Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants

*Evelyn Asante-Kwatia, Abraham Yeboah Mensah and Michael Frimpong Baidoo*

### **Abstract**

Medicinal plants continue to be used in various cultures of the world as safe therapeutic agents against various issues including pain and inflammation which underlie almost every disease process. In Ghanaian traditional medicine, various parts of several plants have been used alone or in combination of therapies for the treatment of various painful inflammatory conditions. In this chapter, the anti-inflammatory and analgesic (antinociceptive) properties of selected medicinal plants from Ghana are reviewed. Evidence of pharmacological activities of crude extracts and fractions in *in-vitro* and *in-vivo* models, bioactive anti-inflammatory and antinociceptive compounds isolated as well as possible mechanisms of anti-inflammatory and antinociceptive action are discussed.

**Keywords:** inflammation, nociception, analgesia, herbal medicine, Ghana

### **1. Introduction**

Inflammation is a complex defensive and protective response of living tissues to injury, irritation or infection which is accompanied by typical symptoms of pain, swelling, redness and fever. It is a mechanism by which the body identifies and neutralises noxious stimuli by increasing the blood flow to the site of tissue injured. Inflammation is a defensive mechanism but the complexity of events as well as the mediators released often result in the induction or aggravation of several disease conditions [1, 2]. Painful conditions such as rheumatoid arthritis, osteoarthritis, asthma, inflammatory bowel disease, colitis and hepatitis as well as other chronic diseases including cardiovascular and neurodegenerative diseases are all conditions whose pathophysiology involves inflammation [3]. These diseases impose a huge social and economic burden on individual victims, their families, and societies as a whole. Moreover, they can cause disability, impairing the social function of people, reducing their quality of life and sometimes resulting in death [2]. Millions of people suffering from different types of painful inflammatory conditions wish to find effective interventions with fewer or no side effects [4].

### **1.1 Current drugs for the treatment of inflammation and pain and their major side effects**

The range of anti-inflammatory and analgesic agents currently available all work to relieve pain, reduce inflammation, and slow down or stop tissue damage. These include non-steroidal anti-inflammatory drugs (NSAIDs), disease modifying antirheumatic drugs (DMARDs), opioids and corticosteroids. Some antidepressants and anti-convulsants have also been shown to increase patients' threshold to pain [5].

NSAIDS such as diclofenac, ibuprofen and aspirin act by blocking certain stages of the arachidonic acid pathway, specifically by inhibiting lipoxygenase (LOX) and cyclooxygenase enzymes (COX-1 and COX-2) responsible for converting arachidonic acid to prostaglandins (PGs). Though effective, NSAIDS are associated with major adverse effects such as gastrointestinal ulceration, intestinal perforation, cardiovascular risks, hepatotoxicity and renal failure after long term use [6].

DMARDs such as methotrexate, sulfasalazine, gold compounds and penicillamine slow the progression of joint destruction in chronic inflammatory conditions like arthritis but are reported to cause kidney failure, skin reactions, liver problems and gastrointestinal side effects [7].

Corticosteroids such as prednisone, cortisone and methylprednisolone act by inhibiting the action of phospholipase A2 which subsequently blocks the biosynthesis of inflammatory mediators such as prostaglandins and leukotrienes. Adverse effects such as delayed wound healing, hypertension, fluid retention, weight gain and osteoporosis are reported [8].

Opioids such as morphine, codeine and pethidine are very effective centrally working analgesics which increase the threshold of pain at the spinal level. These are associated with unwanted behavioural tendencies such as physical dependence, development of tolerance and respiratory depression [9].

#### **1.2 Anti-inflammatory and analgesic agents extracted from medicinal plants**

The adverse effects of most currently used orthodox drugs for the management of painful inflammatory conditions give a strong motivation for researchers to search for other appropriate and effective treatment [10]. Through this search, drugs of plant origin have attracted much attention due to their wide acceptance, availability, reported effectiveness and safety. The discovery of the anti-inflammatory agent salicin and subsequently, aspirin from *Salix fragilis* was a significant evidence to affirm the ability of plants to produce anti-inflammatory compounds [11]. Other plant products such as capsaicin (*Capsicum annum*), curcumin (*Curcuma longa*) and frankincense (*Boswellia serrata*) have been effectively utilised as adjuncts in the treatment of inflammatory conditions and pain [12–14]. Apart from being potent, these products have an added advantage of causing no significant adverse effect or toxicity to liver and kidney cells like other synthetic agents. Medicinal plants are therefore considered as sources of anti-inflammatory and analgesic agents and as practicable alternatives to conventional medicines [15].

#### **1.3 Experimental methods used for screening anti-inflammatory and antinociceptive activities of herbal extracts**

Based on the symptoms of inflammation, several *in-vivo* and *in-vitro* screening methods have been employed to evaluate the anti-inflammatory activity of plant extracts and natural compounds.

**5**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

have been used. To induce inflammation, phlogistic substances or irritants such as carrageenan, mustard, dextran, egg-white, yeast, zymosan-LOX, serotonin, histamine, kaolin, etc. are employed. Some applicable methods described in literature are the carrageenan-induced paw oedema in rats or chicks, croton-oil or oxazolone-induced ear oedema in mice, UV erythema in guinea pigs, granuloma pouch technique and pleurisy in mice [16]. Adjuvant-induced and collagen-induced arthritis models are also efficient in chronic inflammation studies. *In-vitro* methods have mainly focused on the inhibition of the activation of local inflammatory mediators such as leukotrienes [tumour necrosis factor alpha (TNF-α), interleukins (IL-6, IL-1β)], prostaglandins (PGE2), prostacyclin, thromboxane A2, interferon-λ (IFN-λ), inducible nitric oxide synthase (iNOS) and reactive oxygen species. The level of these mediators at the inflamed site is measured and compared to control groups [17, 18]. Other *in-vitro* methods include human red blood cell stabilisation

To determine the antinociceptive effects of herbal extracts, chemically-induced (formalin and acetic acid-induced writhing test) and thermal-induced pain models (hot plate, tail immersion, tail flick, Hargreaves paw withdrawal methods) in

Like other developing countries, Ghana continues to search for more effective and appropriate ways of providing the health needs of its developing populace. Generally, the high cost of Western therapeutic medications and additionally their unavailability to the rural communities has prompted a high interest for herbal medicines [20]. In this regard, intensive efforts are being made to explore plants that might be of therapeutic significance to the Ghanaian community. Several reports cutting across the boundaries of botany, medicine and pharmacy have highlighted the use of different plants alone or in combination therapies for the

Considering the evolving interest in studying traditional systems of healthcare and exploiting the potential of natural products for future drug development, this communication presents a compilation of data on plants with promising anti-inflammatory and analgesic activity with special emphasis on plants found in Ghana. Their pharmacological action, anti-inflammatory or analgesic constituents and possible mechanisms of actions are hereby discussed. It is envisioned that this information will be helpful to the indigenes for their primary healthcare and for researchers, to further identify the active chemical constituents and mechanisms responsible for the analgesic and anti-inflammatory potential of these plants [24].

**1.5 Methods used for identifying herbal materials with anti-inflammatory and** 

Electronic databases including PubMed, SciFinder and Google Scholar were employed in the search for medicinal plants with reported anti-inflammatory and analgesic activities collected from various parts of Ghana. The inclusion criteria were that (i) plant should be used in Ghanaian traditional medicine for treatment of inflammatory condition or pain; (ii) validated *in-vitro* and *in-vivo* models for screening anti-inflammatory and antinociceptive activity were employed; (iii) the right botanical names, plant parts used, types of extracts prepared, active constituents and mechanisms of action if identified were mentioned. Consideration was also given to plants with significant activity differences with

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

and protein denaturation assays.

experimental animals are commonly used [19].

**1.4 Use of herbal medicine in Ghana**

treatment diseases [21–23].

**analgesic activities**

reference to control groups.

To investigate the anti-inflammatory activity of plant extracts against acute and chronic inflammation *in-vivo*, oedema, granuloma and arthritis models

#### *Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants DOI: http://dx.doi.org/10.5772/intechopen.90154*

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

**side effects**

and gastrointestinal side effects [7].

and osteoporosis are reported [8].

development of tolerance and respiratory depression [9].

**1.1 Current drugs for the treatment of inflammation and pain and their major** 

The range of anti-inflammatory and analgesic agents currently available all work to relieve pain, reduce inflammation, and slow down or stop tissue damage. These include non-steroidal anti-inflammatory drugs (NSAIDs), disease modifying antirheumatic drugs (DMARDs), opioids and corticosteroids. Some antidepressants and anti-convulsants have also been shown to increase patients' threshold to pain [5]. NSAIDS such as diclofenac, ibuprofen and aspirin act by blocking certain stages of the arachidonic acid pathway, specifically by inhibiting lipoxygenase (LOX) and cyclooxygenase enzymes (COX-1 and COX-2) responsible for converting arachidonic acid to prostaglandins (PGs). Though effective, NSAIDS are associated with major adverse effects such as gastrointestinal ulceration, intestinal perforation, cardiovascular risks, hepatotoxicity and renal failure after long term use [6]. DMARDs such as methotrexate, sulfasalazine, gold compounds and penicillamine slow the progression of joint destruction in chronic inflammatory conditions like arthritis but are reported to cause kidney failure, skin reactions, liver problems

Corticosteroids such as prednisone, cortisone and methylprednisolone act by inhibiting the action of phospholipase A2 which subsequently blocks the biosynthesis of inflammatory mediators such as prostaglandins and leukotrienes. Adverse effects such as delayed wound healing, hypertension, fluid retention, weight gain

Opioids such as morphine, codeine and pethidine are very effective centrally working analgesics which increase the threshold of pain at the spinal level. These are associated with unwanted behavioural tendencies such as physical dependence,

**1.2 Anti-inflammatory and analgesic agents extracted from medicinal plants**

The adverse effects of most currently used orthodox drugs for the management of painful inflammatory conditions give a strong motivation for researchers to search for other appropriate and effective treatment [10]. Through this search, drugs of plant origin have attracted much attention due to their wide acceptance, availability, reported effectiveness and safety. The discovery of the anti-inflammatory agent salicin and subsequently, aspirin from *Salix fragilis* was a significant evidence to affirm the ability of plants to produce anti-inflammatory compounds [11]. Other plant products such as capsaicin (*Capsicum annum*), curcumin (*Curcuma longa*) and frankincense (*Boswellia serrata*) have been effectively utilised as adjuncts in the treatment of inflammatory conditions and pain [12–14]. Apart from being potent, these products have an added advantage of causing no significant adverse effect or toxicity to liver and kidney cells like other synthetic agents. Medicinal plants are therefore considered as sources of anti-inflammatory and analgesic

agents and as practicable alternatives to conventional medicines [15].

**antinociceptive activities of herbal extracts**

extracts and natural compounds.

**1.3 Experimental methods used for screening anti-inflammatory and** 

Based on the symptoms of inflammation, several *in-vivo* and *in-vitro* screening methods have been employed to evaluate the anti-inflammatory activity of plant

To investigate the anti-inflammatory activity of plant extracts against acute and chronic inflammation *in-vivo*, oedema, granuloma and arthritis models

**4**

have been used. To induce inflammation, phlogistic substances or irritants such as carrageenan, mustard, dextran, egg-white, yeast, zymosan-LOX, serotonin, histamine, kaolin, etc. are employed. Some applicable methods described in literature are the carrageenan-induced paw oedema in rats or chicks, croton-oil or oxazolone-induced ear oedema in mice, UV erythema in guinea pigs, granuloma pouch technique and pleurisy in mice [16]. Adjuvant-induced and collagen-induced arthritis models are also efficient in chronic inflammation studies. *In-vitro* methods have mainly focused on the inhibition of the activation of local inflammatory mediators such as leukotrienes [tumour necrosis factor alpha (TNF-α), interleukins (IL-6, IL-1β)], prostaglandins (PGE2), prostacyclin, thromboxane A2, interferon-λ (IFN-λ), inducible nitric oxide synthase (iNOS) and reactive oxygen species. The level of these mediators at the inflamed site is measured and compared to control groups [17, 18]. Other *in-vitro* methods include human red blood cell stabilisation and protein denaturation assays.

To determine the antinociceptive effects of herbal extracts, chemically-induced (formalin and acetic acid-induced writhing test) and thermal-induced pain models (hot plate, tail immersion, tail flick, Hargreaves paw withdrawal methods) in experimental animals are commonly used [19].

#### **1.4 Use of herbal medicine in Ghana**

Like other developing countries, Ghana continues to search for more effective and appropriate ways of providing the health needs of its developing populace. Generally, the high cost of Western therapeutic medications and additionally their unavailability to the rural communities has prompted a high interest for herbal medicines [20]. In this regard, intensive efforts are being made to explore plants that might be of therapeutic significance to the Ghanaian community. Several reports cutting across the boundaries of botany, medicine and pharmacy have highlighted the use of different plants alone or in combination therapies for the treatment diseases [21–23].

Considering the evolving interest in studying traditional systems of healthcare and exploiting the potential of natural products for future drug development, this communication presents a compilation of data on plants with promising anti-inflammatory and analgesic activity with special emphasis on plants found in Ghana. Their pharmacological action, anti-inflammatory or analgesic constituents and possible mechanisms of actions are hereby discussed. It is envisioned that this information will be helpful to the indigenes for their primary healthcare and for researchers, to further identify the active chemical constituents and mechanisms responsible for the analgesic and anti-inflammatory potential of these plants [24].

#### **1.5 Methods used for identifying herbal materials with anti-inflammatory and analgesic activities**

Electronic databases including PubMed, SciFinder and Google Scholar were employed in the search for medicinal plants with reported anti-inflammatory and analgesic activities collected from various parts of Ghana. The inclusion criteria were that (i) plant should be used in Ghanaian traditional medicine for treatment of inflammatory condition or pain; (ii) validated *in-vitro* and *in-vivo* models for screening anti-inflammatory and antinociceptive activity were employed; (iii) the right botanical names, plant parts used, types of extracts prepared, active constituents and mechanisms of action if identified were mentioned. Consideration was also given to plants with significant activity differences with reference to control groups.

### **2. Plants with anti-inflammatory and analgesic activities from Ghana**

#### **2.1** *Albizia zygia* **(DC.) J.F. Macbr. (***Leguminosae-Mimosoideae***)**

*Albizia zygia* is a medium-sized ornamental shade tree widely distributed in secondary forest and semi-deciduous forest zones of West and East Africa. It grows up to about 30 m tall, has a branchless cylindrical bole with a greenish-grey smooth outer bark and an orange-brown fibrous inner bark. It has alternate bipinnately compound leaves and bears oblong flat pods. It is commonly known as the West African walnut and locally called '*okuro*' in Ghana (Akan). The leaf infusion is used for the treatment of lumbago, fever, waist pain and sexually transmitted infection. The bark decoction is administered to treat respiratory tract disease, malaria fever, constipation and worm infestation. The crushed bark is applied topically to treat yaws, heal wounds and toothache [25].

In previous studies, the leaves and roots were evaluated for their analgesic properties in animal models. Oral administration of the 70% ethanolic leaf extract in rats caused a significant reduction in both neurogenic and inflammatory phases of formalin-induced paw licking with maximal inhibition of 67.81 ± 8.73% and 72.85 ± 12.74% respectively [26]. The hydro-alcoholic root extract also caused a significant diminishing of acetic acid-induced visceral pain, formalin-induced paw pain, thermal and carrageenan-induced mechanical hyperalgesia in animals *via* opioidergic, adenosinergic and muscarinic cholinergic mechanisms [27].

To validate its anti-inflammatory effects, the hydro-alcoholic root extract was evaluated in carrageenan-induced paw oedema and caused a significant reduction of paw oedema in cockerels. The extract was found to increase the expression of endogenous antioxidants such as superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) as well as reduced the action of myeloperoxidase (MPO) and malondialdehyde (MDA) levels at the inflamed site [26].

#### **2.2** *Anopyxis klaineana* **(Pierre) Engl. (***Rhizophoraceae***)**

*A. klaineana* is a medium sized to large tree found in the evergreen and semideciduous forest of tropical Africa. It grows up to about 50 m tall, has a branchless, cylindrical bole with longitudinally fissured greyish-brown outer bark and a thick pale orange inner bark. It has simple leathery, glabrous leaves which occur in whorls of 3–4 and bears greenish-white hairy flowers. *A. klaineana* is locally traded as '*kokoti(e)*' in Ghana and '*bodioa*' in Cote d'Ivoire. The stem bark decoction is used to treat joint pain, gonorrhoea, skin and respiratory tract infection, pneumonia, bronchitis and malaria. Its leaves are also applied as a poultice to heal wounds [28].

The anti-inflammatory activity of the stem bark of *A. klaineana* was evaluated in previous studies. Various solvent extracts including the petroleum ether, ethyl acetate and methanol extracts showed anti-inflammatory activity in a time and dose-dependent manner, by suppressing carrageenan-induced foot pad swelling in chicks. A tetranortriterpenoid called methyl angolensate was isolated as the major constituent of the stem bark and showed anti-inflammatory activity by significantly suppressing foot pad oedema in chicks with an ED50 of 4.05 ± 0.0034 [29]. In another study, a tirucallane triterpenoid isolated from the stem bark namely 3,23-dioxotirucalla-7,24-dien-21-oic acid, exhibited remarkable anti-inflammatory activity in a PGE2 competitive inhibition immunoassay with an IC50 value of 3.63 μM which was comparable to the positive control, cortisone (IC50 = 2.59 μM). Methyl angolensate further demonstrated remarkable competitive inhibition of PGE2 with an IC50 of 10.23 μM confirming its *in-vivo* anti-inflammatory effect [30].

**7**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

*C. procera* is a medium-large sized bushy shrub which grows on coarse, sandy or alkaline soils of West and East Africa also found in the Indian Ocean islands and the north of South Africa. The plant can be identified by decussate, broadly ovate, leathery leaves and bears purple flowers with erect lobes [31]. It is locally called '*mpatu-asa*' (Akan) in Ghana. In traditional medicine, the bark decoction is used for the treatment of rheumatism, arthritis, headache and general body pain. The latex from leafy twigs and flowers is used for treating conjunctivitis, nasopharyngeal infection, tooth ache, wound healing, and vermifuge. The root bark decoction is used for treating cutaneous and subcutaneous skin infection and yaws [32, 33]. A study was conducted to determine the anti-inflammatory effect of the alcoholic extract of *C. procera* leaf in *in-vitro* models including the heat-induced haemolysis, hypotonic-induced haemolysis, albumin denaturation and the bovine serum albumin assay. The 70% alcoholic leaf extract at 1000 μg/mL significantly demonstrated anti-inflammatory effect by stabilising human red blood cells exposed to heat (69.24% inhibition) and hypotonic solution (85.09% inhibition). The extract prevented denaturation of protein (albumin) as well as bovine serum by 87.8% and 96.86% respectively. In *in-vivo* studies, the extract caused significant reduction of carrageenan-induced paw oedema in both acute and chronic

*C. erythrocarpos* is a climbing shrub distributed in the coastal scrubs and inlands

To validate the analgesic effect of *C. erythrocarpos*, the 70% ethanol extract of the root, stem bark and leaf were investigated in the formalin-induced nociception, hot plate and acetic acid-induced writhing assays in mice and rats. The root extract (100 mg/kg *p.o.*) was found to significantly and dose-dependently reduce pain in the early and late phases of formalin-induced pain by 47.54 ± 5.65% and 80.01 ± 3.77% respectively via interaction with adenosinergic receptors [36]. In other studies, the leaf extract at 200 mg/kg *p.o.* showed significant analgesic effects by reducing acetic acid writhing by 27.43% and increasing the pain threshold in the

To validate its anti-inflammatory effect in both acute and chronic inflammation, the 70% alcoholic root extract was investigated in the carrageenan-induced paw oedema and Freund's adjuvant-induced arthritis models respectively. The extract at 30 mg/kg *p.o.* caused marked reduction in foot oedema by 48.86 ± 20.41% and significantly reduced knee joint swelling in arthritis by 34.19 ± 15.73%. The extract prevented systemic spread of inflammation from ipsilateral to contralateral limbs [38]. In another study, the leaves, stem bark and roots also demonstrated marked anti-arthritic activity by reducing rat paw volumes in the Complete Freund's adjuvant model with ED50 values (mg/kg) of 182.5, 181.5

of many African countries and commonly referred to as 'salt bush'. The plant is densely thorny and branched with re-curved hooks, growing up to about 6m in height. Its bears green elliptical leaves, which are alternately arranged [35]. The roots are used in traditional medicine for the management of rheumatism and arthritis. Other plant parts also find use in the treatment of eye and ear infection, fever, epilepsy and as aphrodisiac. The powdered root is used at the Center for Scientific Research in to Plant Medicine (CSRPM), Mampong, Ghana for the

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

inflammation [34].

management of arthritis [21].

hot plate assay by 184.5% [37].

and 36.4 respectively [39].

**2.3** *Calotropis procera* **(Ait) f. (***Apocynaceae***)**

**2.4** *Capparis erythrocarpos* **Isert (***Capparaceae***)**

### **2.3** *Calotropis procera* **(Ait) f. (***Apocynaceae***)**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

yaws, heal wounds and toothache [25].

malondialdehyde (MDA) levels at the inflamed site [26].

**2.2** *Anopyxis klaineana* **(Pierre) Engl. (***Rhizophoraceae***)**

**2. Plants with anti-inflammatory and analgesic activities from Ghana**

*Albizia zygia* is a medium-sized ornamental shade tree widely distributed in secondary forest and semi-deciduous forest zones of West and East Africa. It grows up to about 30 m tall, has a branchless cylindrical bole with a greenish-grey smooth outer bark and an orange-brown fibrous inner bark. It has alternate bipinnately compound leaves and bears oblong flat pods. It is commonly known as the West African walnut and locally called '*okuro*' in Ghana (Akan). The leaf infusion is used for the treatment of lumbago, fever, waist pain and sexually transmitted infection. The bark decoction is administered to treat respiratory tract disease, malaria fever, constipation and worm infestation. The crushed bark is applied topically to treat

In previous studies, the leaves and roots were evaluated for their analgesic properties in animal models. Oral administration of the 70% ethanolic leaf extract in rats caused a significant reduction in both neurogenic and inflammatory phases of formalin-induced paw licking with maximal inhibition of 67.81 ± 8.73% and 72.85 ± 12.74% respectively [26]. The hydro-alcoholic root extract also caused a significant diminishing of acetic acid-induced visceral pain, formalin-induced paw pain, thermal and carrageenan-induced mechanical hyperalgesia in animals *via* opioidergic, adenosinergic and muscarinic cholinergic mechanisms [27].

To validate its anti-inflammatory effects, the hydro-alcoholic root extract was evaluated in carrageenan-induced paw oedema and caused a significant reduction of paw oedema in cockerels. The extract was found to increase the expression of endogenous antioxidants such as superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) as well as reduced the action of myeloperoxidase (MPO) and

*A. klaineana* is a medium sized to large tree found in the evergreen and semideciduous forest of tropical Africa. It grows up to about 50 m tall, has a branchless, cylindrical bole with longitudinally fissured greyish-brown outer bark and a thick pale orange inner bark. It has simple leathery, glabrous leaves which occur in whorls of 3–4 and bears greenish-white hairy flowers. *A. klaineana* is locally traded as '*kokoti(e)*' in Ghana and '*bodioa*' in Cote d'Ivoire. The stem bark decoction is used to treat joint pain, gonorrhoea, skin and respiratory tract infection, pneumonia, bronchitis and malaria. Its leaves are also applied as a poultice to heal wounds [28]. The anti-inflammatory activity of the stem bark of *A. klaineana* was evaluated in previous studies. Various solvent extracts including the petroleum ether, ethyl acetate and methanol extracts showed anti-inflammatory activity in a time and dose-dependent manner, by suppressing carrageenan-induced foot pad swelling in chicks. A tetranortriterpenoid called methyl angolensate was isolated as the major constituent of the stem bark and showed anti-inflammatory activity by significantly suppressing foot pad oedema in chicks with an ED50 of 4.05 ± 0.0034 [29]. In another study, a tirucallane triterpenoid isolated from the stem bark namely 3,23-dioxotirucalla-7,24-dien-21-oic acid, exhibited remarkable anti-inflammatory activity in a PGE2 competitive inhibition immunoassay with an IC50 value of 3.63 μM which was comparable to the positive control, cortisone (IC50 = 2.59 μM). Methyl angolensate further demonstrated remarkable competitive inhibition of PGE2 with an IC50 of 10.23 μM confirming its *in-vivo* anti-inflammatory effect [30].

**2.1** *Albizia zygia* **(DC.) J.F. Macbr. (***Leguminosae-Mimosoideae***)**

**6**

*C. procera* is a medium-large sized bushy shrub which grows on coarse, sandy or alkaline soils of West and East Africa also found in the Indian Ocean islands and the north of South Africa. The plant can be identified by decussate, broadly ovate, leathery leaves and bears purple flowers with erect lobes [31]. It is locally called '*mpatu-asa*' (Akan) in Ghana. In traditional medicine, the bark decoction is used for the treatment of rheumatism, arthritis, headache and general body pain. The latex from leafy twigs and flowers is used for treating conjunctivitis, nasopharyngeal infection, tooth ache, wound healing, and vermifuge. The root bark decoction is used for treating cutaneous and subcutaneous skin infection and yaws [32, 33].

A study was conducted to determine the anti-inflammatory effect of the alcoholic extract of *C. procera* leaf in *in-vitro* models including the heat-induced haemolysis, hypotonic-induced haemolysis, albumin denaturation and the bovine serum albumin assay. The 70% alcoholic leaf extract at 1000 μg/mL significantly demonstrated anti-inflammatory effect by stabilising human red blood cells exposed to heat (69.24% inhibition) and hypotonic solution (85.09% inhibition). The extract prevented denaturation of protein (albumin) as well as bovine serum by 87.8% and 96.86% respectively. In *in-vivo* studies, the extract caused significant reduction of carrageenan-induced paw oedema in both acute and chronic inflammation [34].

#### **2.4** *Capparis erythrocarpos* **Isert (***Capparaceae***)**

*C. erythrocarpos* is a climbing shrub distributed in the coastal scrubs and inlands of many African countries and commonly referred to as 'salt bush'. The plant is densely thorny and branched with re-curved hooks, growing up to about 6m in height. Its bears green elliptical leaves, which are alternately arranged [35]. The roots are used in traditional medicine for the management of rheumatism and arthritis. Other plant parts also find use in the treatment of eye and ear infection, fever, epilepsy and as aphrodisiac. The powdered root is used at the Center for Scientific Research in to Plant Medicine (CSRPM), Mampong, Ghana for the management of arthritis [21].

To validate the analgesic effect of *C. erythrocarpos*, the 70% ethanol extract of the root, stem bark and leaf were investigated in the formalin-induced nociception, hot plate and acetic acid-induced writhing assays in mice and rats. The root extract (100 mg/kg *p.o.*) was found to significantly and dose-dependently reduce pain in the early and late phases of formalin-induced pain by 47.54 ± 5.65% and 80.01 ± 3.77% respectively via interaction with adenosinergic receptors [36]. In other studies, the leaf extract at 200 mg/kg *p.o.* showed significant analgesic effects by reducing acetic acid writhing by 27.43% and increasing the pain threshold in the hot plate assay by 184.5% [37].

To validate its anti-inflammatory effect in both acute and chronic inflammation, the 70% alcoholic root extract was investigated in the carrageenan-induced paw oedema and Freund's adjuvant-induced arthritis models respectively. The extract at 30 mg/kg *p.o.* caused marked reduction in foot oedema by 48.86 ± 20.41% and significantly reduced knee joint swelling in arthritis by 34.19 ± 15.73%. The extract prevented systemic spread of inflammation from ipsilateral to contralateral limbs [38]. In another study, the leaves, stem bark and roots also demonstrated marked anti-arthritic activity by reducing rat paw volumes in the Complete Freund's adjuvant model with ED50 values (mg/kg) of 182.5, 181.5 and 36.4 respectively [39].

#### **2.5** *Cassia sieberiana* **D.C. (***Caesalpinaceae***)**

*C. sieberiana* is a tropical woody shrub found growing in the bushy savannahs and coastal shrubs of many African countries. The plant grows up to about 20 m tall, has a short twisted bole, with a greyish-brown fissured bark. It has spirally arranged paripinnately compound leaves which bear bright yellow flowers and dehiscent pods as fruits. The entire plant is purgative and diuretic. The root decoction and leaf infusions are used as pain reliever in rheumatism and arthritis, for treatment of ear infection, skin disease, malaria fever, gastrointestinal infection, oedema, sexually transmitted infection, as laxative and vermifuge. The boiled and squeezed fresh leaves are applied topically to heal wounds, pleurisy and boils [25, 40].

A study conducted to investigate the analgesic effects of the aqueous and ethyl acetate root extracts indicated that in the hot plate assay, the aqueous extract attenuated hyperalgesia in a dose-dependent manner with an ED50 of 9.7 ± 3.9 mg/kg. The ethyl acetate fraction also showed antinociceptive activity in the formalin-induced nociception, yeast induced hyperalgesia, hot plate and acetic acid writhing tests. The analgesic effect was significantly blocked by Naloxone, atropine and theophylline indicating interactions with the opioidergic, muscarinic cholinergic or adenosinergic pathways [41, 42].

*In-vivo* study detected that the ethyl acetate extract of *C. sieberiana* root exhibits anti-inflammatory activity by reducing carrageenan-induced foot oedema in chicks [42]. Furthermore, the 70% ethanolic root extracts dose-dependently attenuated *Mycobacterium tuberculosis*-carrageenan-induced inflammation in the rats. Serum levels of IL-1α, IL-6 and TNF-α were reduced with increasing levels of IL-10 suggesting that the anti-inflammatory activity of the root bark extract may be as a result of its immune-modulatory effects *via* interactions with these pro-inflammatory mediators [43].

#### **2.6** *Commelina diffusa* **Burm. f. (***Commelinaceae***)**

*C. diffusa* is a perennial herb distributed in tropical African countries including Ghana, Nigeria, Ivory Coast, Gabon and Congo. The plant is a smooth and sparsely hairy herb with mucilaginous leaves and creeping stems which ascends above and roots at the nodes. It is commonly called 'climbing day flower'. The Akans in Ghana fancifully refer to it as '*Nyame bewu ansa na mawu*' meaning '*God will die before I die*' alluding to its tenacity to life. In Ghana and Nigeria, the pounded leaves are applied topically to boils and swollen glands and as a rubefacient to relief pain in rheumatism and arthritis. Other reported uses include for the treatment of skin abscess, wound, gonorrhoea, ear infection and for the relief of severe menstrual pain [44].

To evaluate its anti-inflammatory effect, the 70% ethanolic leaf extract was investigated in the carrageenan-induced foot pad oedema in chicks. The extract (30, 100 and 300 mg/kg *p.o.*) showed a dose-dependent inhibition of foot pad oedema with the maximum inhibition of 43.55% at 300 mg/kg confirming it antiinflammatory effects [44].

#### **2.7** *Erythrophleum ivoren***se (A Chev.) (***Fabaceae***)**

*E. ivorense* is a large tree widely distributed in the evergreen primary and secondary forests of tropical Africa. It grows to about 40 m tall, with a cylindrical bole, sometimes fluted at the base. It is called by names like 'forest ordeal tree', 'red water tree' and 'sasswood tree' in West African countries. Among the Akan tribe in Ghana, *E. ivorense* is known as '*potrodum*'. The stem-bark and roots are usually

**9**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

employed in the treatment of epilepsy, emesis, pain, oedema, constipation and

induced foot pad oedema better than the standard drug diclofenac [46].

hyperalgesia in rats was produced by the leaf extract [49].

bark also exhibited anti-inflammatory activities [52].

**2.9** *Glyphaea brevis* **(Spreng) Monachino (***Tiliaceae***)**

constipation, chest pain and gastrointestinal infection [53].

lactically and therapeutically [55].

The hydro-alcoholic stem bark and leaf extracts at 30–300 mg/kg, *p.o.* dose-dependently inhibited carrageenan-induced foot oedema with ED50s of 50.65 ± 0.012 and 46.05 ± 12.3 respectively [50]. Moreover, the leaf extract significantly reduced the arthritic oedema in ipsilateral paws of rats with a maximal inhibition of 34.46 ± 11.42% and significantly prevented the systemic spread to the contralateral paws [51]. Furanocoumarins namely bergapten, oxypeucedanin hydrate and the sterolin, sitosterol-3-O-β-d-glucopyranoside isolated from the stem

*G. brevis* is a medium sized spreading climber usually found growing in forest re-growths, rocky savannahs and swampy areas of tropical Africa. It possesses straggling sparsely stellate branchlets, which bear ovate-oblong leaves and lemonyellow flowers. Its fruits are spindle-shaped and brown in colour with irregularly ellipsoid seeds. The leaves are used to treat dyspepsia, gastric ulcer, oedema, pain and worm infestation. The root decoction is used to treat male sexual impotence,

The anti-inflammatory effects of the 70% ethanol extracts of the leaves and stem bark were investigated by the carrageenan induced foot pad oedema method. The extracts exhibited potent anti-inflammatory activity in doses of 30, 100 and 300 mg/kg *p.o.*, by reducing foot oedema with similar potencies at ED50s ~ 21.00 mg/kg [54]. In another study, oral administration of the 70% ethanol extract of the stem bark exerted inhibitory effects on carrageenan-induced paw oedema, systemic anaphylaxis and chronic inflammation in the Freund's adjuvant-induced arthritis models. The effect was significant when the extract was given both prophy-

The carrageenan-induced foot pad oedema in chick was used to evaluate the antiinflammatory activity of the roots of *E. ivorense*. The 70% alcoholic root extract suppressed foot pad oedema in a time and dose-dependent manner. Three constituents, a casein type diterpene namely erythroivorensin A, betulininc acid and the flavonoid, eriodictyol isolated from the roots exhibited significant reduction of carrageenan-

*F. exasperata*, commonly known as 'sand paper tree', is a deciduous, shrub growing up to about 30 m tall. It has a buttressed bole with a pale grey-green outer bark and creamy-white inner bark which exudes a clear, viscid sap when damaged. It has alternate simple pubescent leaves which are elliptical in shape. In Ghana, it is locally called '*onyankyerεn*' (Akan), '*nyadεlε*' (Nzema) or *nyadkese* (Ga). The plant is used in folk medicine for the treatment of sprain, arthritis, rheumatism, intestinal and stomach infection, high blood pressure, abscesses and respiratory tract disease [47]. The analgesic activity of the 70% alcoholic leaf extract was investigated in murine models. The extract elicited a dose-dependent significant antinociceptive effect in the formalin-induced nociception assay through interactions with adenosinergic and opioidergic pathways [48]. The leaf extract also caused significant reduction in acute carrageenan-kaolin-induced muscle hyperalgesia (ED50 = 31.23 ± 11.91). Significant attenuation of chronic muscle hyperalgesia in both ipsilateral and contralateral paws and total reversal of the chronic muscle

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

**2.8** *Ficus exasperata* **Vahl (***Moraceae***)**

worm infestation [45].

#### *Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants DOI: http://dx.doi.org/10.5772/intechopen.90154*

employed in the treatment of epilepsy, emesis, pain, oedema, constipation and worm infestation [45].

The carrageenan-induced foot pad oedema in chick was used to evaluate the antiinflammatory activity of the roots of *E. ivorense*. The 70% alcoholic root extract suppressed foot pad oedema in a time and dose-dependent manner. Three constituents, a casein type diterpene namely erythroivorensin A, betulininc acid and the flavonoid, eriodictyol isolated from the roots exhibited significant reduction of carrageenaninduced foot pad oedema better than the standard drug diclofenac [46].

#### **2.8** *Ficus exasperata* **Vahl (***Moraceae***)**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

*C. sieberiana* is a tropical woody shrub found growing in the bushy savannahs and coastal shrubs of many African countries. The plant grows up to about 20 m tall, has a short twisted bole, with a greyish-brown fissured bark. It has spirally arranged paripinnately compound leaves which bear bright yellow flowers and dehiscent pods as fruits. The entire plant is purgative and diuretic. The root decoction and leaf infusions are used as pain reliever in rheumatism and arthritis, for treatment of ear infection, skin disease, malaria fever, gastrointestinal infection, oedema, sexually transmitted infection, as laxative and vermifuge. The boiled and squeezed fresh

A study conducted to investigate the analgesic effects of the aqueous and ethyl acetate root extracts indicated that in the hot plate assay, the aqueous extract attenuated hyperalgesia in a dose-dependent manner with an ED50 of 9.7 ± 3.9 mg/kg. The ethyl acetate fraction also showed antinociceptive activity in the formalin-induced nociception, yeast induced hyperalgesia, hot plate and acetic acid writhing tests. The analgesic effect was significantly blocked by Naloxone, atropine and theophylline indicating interactions with the opioidergic, muscarinic cholinergic or adenosinergic

*In-vivo* study detected that the ethyl acetate extract of *C. sieberiana* root exhib-

*C. diffusa* is a perennial herb distributed in tropical African countries including Ghana, Nigeria, Ivory Coast, Gabon and Congo. The plant is a smooth and sparsely hairy herb with mucilaginous leaves and creeping stems which ascends above and roots at the nodes. It is commonly called 'climbing day flower'. The Akans in Ghana fancifully refer to it as '*Nyame bewu ansa na mawu*' meaning '*God will die before I die*' alluding to its tenacity to life. In Ghana and Nigeria, the pounded leaves are applied topically to boils and swollen glands and as a rubefacient to relief pain in rheumatism and arthritis. Other reported uses include for the treatment of skin abscess, wound, gonorrhoea, ear infection and for the relief of severe menstrual

To evaluate its anti-inflammatory effect, the 70% ethanolic leaf extract was investigated in the carrageenan-induced foot pad oedema in chicks. The extract (30, 100 and 300 mg/kg *p.o.*) showed a dose-dependent inhibition of foot pad oedema with the maximum inhibition of 43.55% at 300 mg/kg confirming it anti-

*E. ivorense* is a large tree widely distributed in the evergreen primary and secondary forests of tropical Africa. It grows to about 40 m tall, with a cylindrical bole, sometimes fluted at the base. It is called by names like 'forest ordeal tree', 'red water tree' and 'sasswood tree' in West African countries. Among the Akan tribe in Ghana, *E. ivorense* is known as '*potrodum*'. The stem-bark and roots are usually

its anti-inflammatory activity by reducing carrageenan-induced foot oedema in chicks [42]. Furthermore, the 70% ethanolic root extracts dose-dependently attenuated *Mycobacterium tuberculosis*-carrageenan-induced inflammation in the rats. Serum levels of IL-1α, IL-6 and TNF-α were reduced with increasing levels of IL-10 suggesting that the anti-inflammatory activity of the root bark extract may be as a result of its immune-modulatory effects *via* interactions with these

leaves are applied topically to heal wounds, pleurisy and boils [25, 40].

**2.5** *Cassia sieberiana* **D.C. (***Caesalpinaceae***)**

pathways [41, 42].

pro-inflammatory mediators [43].

**2.6** *Commelina diffusa* **Burm. f. (***Commelinaceae***)**

**2.7** *Erythrophleum ivoren***se (A Chev.) (***Fabaceae***)**

**8**

pain [44].

inflammatory effects [44].

*F. exasperata*, commonly known as 'sand paper tree', is a deciduous, shrub growing up to about 30 m tall. It has a buttressed bole with a pale grey-green outer bark and creamy-white inner bark which exudes a clear, viscid sap when damaged. It has alternate simple pubescent leaves which are elliptical in shape. In Ghana, it is locally called '*onyankyerεn*' (Akan), '*nyadεlε*' (Nzema) or *nyadkese* (Ga). The plant is used in folk medicine for the treatment of sprain, arthritis, rheumatism, intestinal and stomach infection, high blood pressure, abscesses and respiratory tract disease [47].

The analgesic activity of the 70% alcoholic leaf extract was investigated in murine models. The extract elicited a dose-dependent significant antinociceptive effect in the formalin-induced nociception assay through interactions with adenosinergic and opioidergic pathways [48]. The leaf extract also caused significant reduction in acute carrageenan-kaolin-induced muscle hyperalgesia (ED50 = 31.23 ± 11.91). Significant attenuation of chronic muscle hyperalgesia in both ipsilateral and contralateral paws and total reversal of the chronic muscle hyperalgesia in rats was produced by the leaf extract [49].

The hydro-alcoholic stem bark and leaf extracts at 30–300 mg/kg, *p.o.* dose-dependently inhibited carrageenan-induced foot oedema with ED50s of 50.65 ± 0.012 and 46.05 ± 12.3 respectively [50]. Moreover, the leaf extract significantly reduced the arthritic oedema in ipsilateral paws of rats with a maximal inhibition of 34.46 ± 11.42% and significantly prevented the systemic spread to the contralateral paws [51]. Furanocoumarins namely bergapten, oxypeucedanin hydrate and the sterolin, sitosterol-3-O-β-d-glucopyranoside isolated from the stem bark also exhibited anti-inflammatory activities [52].

#### **2.9** *Glyphaea brevis* **(Spreng) Monachino (***Tiliaceae***)**

*G. brevis* is a medium sized spreading climber usually found growing in forest re-growths, rocky savannahs and swampy areas of tropical Africa. It possesses straggling sparsely stellate branchlets, which bear ovate-oblong leaves and lemonyellow flowers. Its fruits are spindle-shaped and brown in colour with irregularly ellipsoid seeds. The leaves are used to treat dyspepsia, gastric ulcer, oedema, pain and worm infestation. The root decoction is used to treat male sexual impotence, constipation, chest pain and gastrointestinal infection [53].

The anti-inflammatory effects of the 70% ethanol extracts of the leaves and stem bark were investigated by the carrageenan induced foot pad oedema method. The extracts exhibited potent anti-inflammatory activity in doses of 30, 100 and 300 mg/kg *p.o.*, by reducing foot oedema with similar potencies at ED50s ~ 21.00 mg/kg [54]. In another study, oral administration of the 70% ethanol extract of the stem bark exerted inhibitory effects on carrageenan-induced paw oedema, systemic anaphylaxis and chronic inflammation in the Freund's adjuvant-induced arthritis models. The effect was significant when the extract was given both prophylactically and therapeutically [55].

#### **2.10** *Haematostaphis barteri* **Hook. f. (***Anarcadiaceae***)**

*H. barteri* is a woody plant typical of tropical Africa widely distributed in rocky savanna areas of Ghana, Upper Volta, Nigeria, Cameroon and Sudan. It reaches up to about 8 m high, about 65 cm in girth with a bark that contains a clear gum. It bears characteristic reddish-purple drupes which are edible with an acrid taste [33]. It is commonly called 'blood plum' and in the Upper West region of Ghana where it is locally referred to as '*zimbringa*' (Dagaari). In traditional medicine, the boiled leaves are used to treat malaria. The stem bark decoction is used for the treatment of hepatitis and sleeping sickness, while the roots are used in the treatment of oedema, pain and swelling [56].

The antinociceptive and anti-inflammatory effects of the plant were investigated in previous studies. The aqueous leaf extract significantly blocked the progression of the neurogenic and inflammatory phases of formalin-induced nociception in a dose-dependent manner. The study further revealed that *H. barteri* inhibits nociception in mice by modulating the opioidergic, adrenergic, muscarinic, ATPsensitive K+ channels and adenosinergic nociceptive pathways [57]. Moreover, the aqueous leaf extract inhibited carrageenan, histamine and serotonin-induced rat paw oedema significantly [58].

#### **2.11** *Hilleria latifolia* **(Lam.) H.Walt. (***Phytolaccaceae***)**

*H. latifolia* is a woody perennial herb about 2 m tall, with weak spiky hairs on young branches. It has alternate, simple elliptical leaves, bears several whitish-green sepals and a lens-shaped fruit with a thin wrinkled pericarp. In Ghana it is locally called '*Avegboma*' (Ewe) and '*Anafranaku*' (Akan-Twi) and used for the treatment of arthritis, rheumatism, oedema, gout, worm infestation, parasitic and viral infection of the skin, respiratory and pulmonary disease including asthma [25, 59].

The ethanolic extract of the aerial plant parts of the plant was investigated for analgesic and anti-inflammatory effect *in-vivo*. The extract in doses of 30–300 mg/kg *p.o.* demonstrated remarkable antinociceptive activity in the chemical and thermal-induced pain models. It produced a dose-related analgesic effect and significantly suppressed the development of morphine tolerance after repeated co-administration with morphine [60]. Its analgesic effect is *via* alteration of adenosinergic, muscarinic cholinergic and opioid pathways [61].

The 70% ethanolic extract of the aerial parts also significantly inhibited acute inflammation in the carrageenan-induced foot oedema [61] and significantly reduced poly-arthritic oedema in the ipsilateral paw of rats but was unable to prevent systemic spread to contralateral limbs in the Freund's adjuvant-induced arthritis model [62].

#### **2.12** *Jatropha curcas* **L. (***Euphorbiaceae***)**

*J. curcas* is a shrub or small tree about, 2–5 m tall, with a smooth bark and sparsely lenticellate branches. The leaves are broadly palmate and inflorescences greenish-yellow. At maturity it produces ellipsoidal capsules containing black seeds. The seed oil is used to treat eczema, skin disease and to soothe rheumatic pain. The root powder is topically applied as a paste to treat swelling and inflammatory condition such as gout [63].

In studies of its analgesic activity, the 70% ethanolic root extract (30–300 mg/kg, *p.o.*) significantly inhibited acute and chronic skeletal hyperalgesia induced by 3% kaolin-carrageenan mixture in both ipsilateral and contralateral limbs of rats [64].

**11**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

condition, pain, schistosomiasis, haemorrhoid and toothache [65].

The name of the genus, *Lannea*, originates from the Latin word '*lana*' which translates to 'wool' alluding to the densely hairy young plant parts or possibly to the wool on the roots of some *Lannea* species. The plant occurs in different habitats in Sub-Saharan Africa including Benin, Burkina Faso, Cameroon, Central African Republic, Côte d'Ivoire, Gambia, Ghana, Guinea, Mali, Niger, and Nigeria. It usually grows in wooded savannah, forest edges, bushed grassland, rocky outcrops, and near rivers on sandy soils. It bears berry-like fruits which occur in large clusters and are consumed either fresh or dried. The fruits have a slightly acidic but pleasant taste. In traditional medicine, *L. acida* is used for the treatment of inflammatory

The aqueous stem bark extract was evaluated for anti-inflammatory effect and caused a significant dose-dependent reduction of PGE2-induced rat paw oedema with maximal oedema inhibition of 67.1%. The stem bark extract also inhibited writhing movement in the acetic acid-induced writhing test in mice models [66].

*N. laevis* is a shrubby small to medium sized ornamental tree with several vertically ascending stems usually found growing in the wooded savanna and deciduous forests across tropical Africa. The plant has shiny dark green leaves and bears large terminal purple flowers. *N. laevis* finds use in folk medicine for the treatment of epilepsy, elephantiasis, haemorrhoid, pelvic pain, peptic and skin ulcer, rheuma-

The analgesic and anti-inflammatory activity of the leaves have been inves-

The ethanolic leaf extract significantly and dose-dependently, inhibited carrageenan-induced foot oedema with maximal inhibition of 64.41 ± 11.47% [67]. In another study, the ethanol stem bark extract inhibited the poly-arthritic phase limb swelling in rat adjuvant-induced arthritis by 28.11 ± 2.02% justifying the use of the

*P. hirsuta* is one of the most commonly used species of Commelinaceae. It is a robust perennial herb with lax inflorescences, lateral branches, purplish flowers and black glossy fruits. It is usually found in lowland rain-forest of West Africa. In Ghana it is commonly called '*somenini*' or '*mpentemi*' in Akan, '*sumbe*' in Ewe and '*sombenyin*' in Fante languages. Various parts of the plant are used in traditional medicine for the treatment of general body pain, earache, pelvic pain, piles, tooth

The ethanolic leaf extract of *P. hirsuta* was investigated for its analgesic effect. The extract (30–300 mg/kg *p.o.*) caused a significant increase in tail withdrawal latency by 73.75 ± 14.99%; reversed carrageenan-induced hyperalgesia with a percentage maximum effect of 154.79 ± 15.84%; reduced the number of acetic acid

tigated in several models. At 300 mg/kg *p.o.*, the 70% ethanol leaf extract significantly increased the paw withdrawal latency of mice in a tail immersion (withdrawal) test by 88.45 ± 19.81% indicating decreased sensitivity to pain. The leaf extract further inhibited the neurogenic (54.47 ± 8.60%) and inflammatory phases (83.62 ± 6.03%) of formalin-induced nociception and blocked the effect of carrageenan-induced thermal hyperalgesia by 37.60 ± 7.26% [67]. In another study, the hydro-alcoholic stem bark extract significantly and dose-dependently decreased

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

**2.13** *Lannea acida* **A. Rich (***Anacadiaceae***)**

**2.14** *Newbouldia laevis* **Seem. (***Bignoniaceae***)**

tism and as antidote to snake bite [40].

formalin-induced nociceptive behaviour in rats [68].

stem bark in the management of arthritis [69].

ache, swelling and wound [70].

**2.15** *Palisota hirsuta* **K. Schum (***Commelinaceae***)**

#### **2.13** *Lannea acida* **A. Rich (***Anacadiaceae***)**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

**2.10** *Haematostaphis barteri* **Hook. f. (***Anarcadiaceae***)**

**2.11** *Hilleria latifolia* **(Lam.) H.Walt. (***Phytolaccaceae***)**

pain and swelling [56].

paw oedema significantly [58].

arthritis model [62].

tion such as gout [63].

limbs of rats [64].

**2.12** *Jatropha curcas* **L. (***Euphorbiaceae***)**

sensitive K+

*H. barteri* is a woody plant typical of tropical Africa widely distributed in rocky savanna areas of Ghana, Upper Volta, Nigeria, Cameroon and Sudan. It reaches up to about 8 m high, about 65 cm in girth with a bark that contains a clear gum. It bears characteristic reddish-purple drupes which are edible with an acrid taste [33]. It is commonly called 'blood plum' and in the Upper West region of Ghana where it is locally referred to as '*zimbringa*' (Dagaari). In traditional medicine, the boiled leaves are used to treat malaria. The stem bark decoction is used for the treatment of hepatitis and sleeping sickness, while the roots are used in the treatment of oedema,

The antinociceptive and anti-inflammatory effects of the plant were investigated

channels and adenosinergic nociceptive pathways [57]. Moreover, the

in previous studies. The aqueous leaf extract significantly blocked the progression of the neurogenic and inflammatory phases of formalin-induced nociception in a dose-dependent manner. The study further revealed that *H. barteri* inhibits nociception in mice by modulating the opioidergic, adrenergic, muscarinic, ATP-

aqueous leaf extract inhibited carrageenan, histamine and serotonin-induced rat

*H. latifolia* is a woody perennial herb about 2 m tall, with weak spiky hairs on young branches. It has alternate, simple elliptical leaves, bears several whitish-green sepals and a lens-shaped fruit with a thin wrinkled pericarp. In Ghana it is locally called '*Avegboma*' (Ewe) and '*Anafranaku*' (Akan-Twi) and used for the treatment of arthritis, rheumatism, oedema, gout, worm infestation, parasitic and viral infection of the skin, respiratory and pulmonary disease including asthma [25, 59]. The ethanolic extract of the aerial plant parts of the plant was investigated for

30–300 mg/kg *p.o.* demonstrated remarkable antinociceptive activity in the chemical and thermal-induced pain models. It produced a dose-related analgesic effect and significantly suppressed the development of morphine tolerance after repeated co-administration with morphine [60]. Its analgesic effect is *via* alteration of

The 70% ethanolic extract of the aerial parts also significantly inhibited acute inflammation in the carrageenan-induced foot oedema [61] and significantly reduced poly-arthritic oedema in the ipsilateral paw of rats but was unable to prevent systemic spread to contralateral limbs in the Freund's adjuvant-induced

*J. curcas* is a shrub or small tree about, 2–5 m tall, with a smooth bark and sparsely lenticellate branches. The leaves are broadly palmate and inflorescences greenish-yellow. At maturity it produces ellipsoidal capsules containing black seeds. The seed oil is used to treat eczema, skin disease and to soothe rheumatic pain. The root powder is topically applied as a paste to treat swelling and inflammatory condi-

(30–300 mg/kg, *p.o.*) significantly inhibited acute and chronic skeletal hyperalgesia induced by 3% kaolin-carrageenan mixture in both ipsilateral and contralateral

In studies of its analgesic activity, the 70% ethanolic root extract

analgesic and anti-inflammatory effect *in-vivo*. The extract in doses of

adenosinergic, muscarinic cholinergic and opioid pathways [61].

**10**

The name of the genus, *Lannea*, originates from the Latin word '*lana*' which translates to 'wool' alluding to the densely hairy young plant parts or possibly to the wool on the roots of some *Lannea* species. The plant occurs in different habitats in Sub-Saharan Africa including Benin, Burkina Faso, Cameroon, Central African Republic, Côte d'Ivoire, Gambia, Ghana, Guinea, Mali, Niger, and Nigeria. It usually grows in wooded savannah, forest edges, bushed grassland, rocky outcrops, and near rivers on sandy soils. It bears berry-like fruits which occur in large clusters and are consumed either fresh or dried. The fruits have a slightly acidic but pleasant taste. In traditional medicine, *L. acida* is used for the treatment of inflammatory condition, pain, schistosomiasis, haemorrhoid and toothache [65].

The aqueous stem bark extract was evaluated for anti-inflammatory effect and caused a significant dose-dependent reduction of PGE2-induced rat paw oedema with maximal oedema inhibition of 67.1%. The stem bark extract also inhibited writhing movement in the acetic acid-induced writhing test in mice models [66].

#### **2.14** *Newbouldia laevis* **Seem. (***Bignoniaceae***)**

*N. laevis* is a shrubby small to medium sized ornamental tree with several vertically ascending stems usually found growing in the wooded savanna and deciduous forests across tropical Africa. The plant has shiny dark green leaves and bears large terminal purple flowers. *N. laevis* finds use in folk medicine for the treatment of epilepsy, elephantiasis, haemorrhoid, pelvic pain, peptic and skin ulcer, rheumatism and as antidote to snake bite [40].

The analgesic and anti-inflammatory activity of the leaves have been investigated in several models. At 300 mg/kg *p.o.*, the 70% ethanol leaf extract significantly increased the paw withdrawal latency of mice in a tail immersion (withdrawal) test by 88.45 ± 19.81% indicating decreased sensitivity to pain. The leaf extract further inhibited the neurogenic (54.47 ± 8.60%) and inflammatory phases (83.62 ± 6.03%) of formalin-induced nociception and blocked the effect of carrageenan-induced thermal hyperalgesia by 37.60 ± 7.26% [67]. In another study, the hydro-alcoholic stem bark extract significantly and dose-dependently decreased formalin-induced nociceptive behaviour in rats [68].

The ethanolic leaf extract significantly and dose-dependently, inhibited carrageenan-induced foot oedema with maximal inhibition of 64.41 ± 11.47% [67]. In another study, the ethanol stem bark extract inhibited the poly-arthritic phase limb swelling in rat adjuvant-induced arthritis by 28.11 ± 2.02% justifying the use of the stem bark in the management of arthritis [69].

#### **2.15** *Palisota hirsuta* **K. Schum (***Commelinaceae***)**

*P. hirsuta* is one of the most commonly used species of Commelinaceae. It is a robust perennial herb with lax inflorescences, lateral branches, purplish flowers and black glossy fruits. It is usually found in lowland rain-forest of West Africa. In Ghana it is commonly called '*somenini*' or '*mpentemi*' in Akan, '*sumbe*' in Ewe and '*sombenyin*' in Fante languages. Various parts of the plant are used in traditional medicine for the treatment of general body pain, earache, pelvic pain, piles, tooth ache, swelling and wound [70].

The ethanolic leaf extract of *P. hirsuta* was investigated for its analgesic effect. The extract (30–300 mg/kg *p.o.*) caused a significant increase in tail withdrawal latency by 73.75 ± 14.99%; reversed carrageenan-induced hyperalgesia with a percentage maximum effect of 154.79 ± 15.84%; reduced the number of acetic acid writhing with an ED50 of 80.20 ± 0.58 mg/kg and decreased formalin-induced nociception by 83.46 ± 6.67% and 94.56 ± 4.12% in the early and late phases respectively [71].

In other studies, oral administration of the leaf extract (30–300 mg/kg *p.o.*) resulted in a dose-dependent complete reversal of vincristine-induced neuropathic pain in rats [72]. An ecdysteroid called 20-hydroxyecdysone was isolated from the root and was found to inhibit formalin-induced nociception in rats by 71.39 ± 9.19% and 89.19 ± 3.81% respectively in the early and late phases [73].

The ethanolic root extract (50–400 mg/kg *p.o.*) demonstrated remarkable reduction of carrageenan-induced foot oedema in chicks in both curative (62.52 ± 4.73%) and prophylactic (58.90 ± 11.38%) treatment regimens [74]. Further, the ethanolic leaf extract caused significant reduction in arthritic oedema induced by Freund's adjuvant and prevented the systemic spread of arthritis from the ipsilateral to the contralateral limb [75].

#### **2.16** *Picralima nitida* **(Stapf ) T. Durand & H. Durand (***Apocynaceae***)**

*P. nitida* is a medium sized to large tree which reaches up to 35 m in height with a dense crown, a pale yellow, fine grained inner wood and a cylindrical trunk. The leaves are broadly oblong with hard tiny lateral nerves and bear white flowers with ovoid fruits which turn yellow at maturity. *P. nitida* is widely distributed in the deciduous forests of West and Central Africa. In Ghana, the seeds are locally known as '*akuama*' (Asante-Akan) or '*onwema*' (Fante) and are used for the treatment of pain of various aetiologies as well as fever. Other plant parts find use in folk medicine for the treatment of malaria, fever, worm infestation, venereal disease, respiratory tract infection, constipation and jaundice [76].

Investigation of the analgesic effects of seeds collected from Ghana established that the aqueous seed extract possessed significant antinociceptive effect in murine models tested by the hot plate assay. Indole alkaloids isolated from the seeds, namely akuammidine, akuammine, akuammicine, akuammigine and pseudoakuammigine also exhibited potent analgesic effects in an isolated tissue and radio-ligand binding assay, demonstrating varying degrees of agonist and antagonist activity at *μ*-, *δ*-, and *κ*-opioid receptors [77, 78].

In anti-inflammatory studies, the hydro-ethanolic extract of the seeds demonstrated a dose-dependent suppression of paw oedema in the carrageenan-induced paw oedema assay. The extract further showed inhibition of chronic inflammation in rat adjuvant-induced arthritis. The total alkaloidal extract at 75–300 mg/kg *p.o.* caused a significant dose-dependent inhibition of total oedema formation in carrageenan-induced paw oedema assay and reduced adjuvant-induced knee joint swelling in rats [79]. Pseudoakuammigine displayed significant dose-dependent suppression of total paw oedema by 82.8 ± 94.6% [78].

#### **2.17** *Phyllanthus muellerianus* **(Kuntze) Exell. (***Euphorbiaceae***)**

*P. muellerianus* is a straggling shrub about 12 m tall with spreading branches and several short axillary shoots dispersed in the deciduous and secondary forests of tropical Africa. It has simple alternate glabrous leaves and clustered whitish-green flowers. The plant bears fleshy six-seeded smooth capsules which are green when young and black at maturity. The fresh twigs are chewed to prevent toothache and also used to treat dysmenorrhea, dropsy, wound, swelling, oedema, tumour, paralysis and epilepsy [80].

The aerial part of the plant was investigated for analgesic and anti-inflammatory effects in various models. Oral administration of the aqueous extract in doses

**13**

haemostatic [86].

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

**2.18** *Secamone afzelii* **(Schult.) K. Schum (***Asclepiadaceae***)**

extract gave a 44.26% inhibition of oedema [85].

**2.19** *Synedrella nodiflora* **(Linn.) Gaertn. (***Asteraceae***)**

of 30, 100, 300 mg/kg, produced significant antinociceptive effect in the acetic acid-induced abdominal writhing and formalin-induced nociception models in rats [81]. The antinociceptive effect of its major constituent geraniin was demonstrated *via* interaction with opioidergic receptors. Geraniin was found to potentiate the antinociceptive effects of diclofenac and morphine when co-administered [82]. In the carrageenan-induced acute inflammation model, the 70% ethanolic extract of the whole plant and 10 mg/kg of its major constituent geraniin significantly reduced paw oedema by 46.75 ± 4.97% and 61.65 ± 6.70% respectively. The extract and geraniin further attenuated arthritis by reducing total limb swelling in the Freund's adjuvant-induced arthritis model. Histomorphological analysis revealed reduced bone damage in both extract and geraniin treated groups [83].

*S. afzelii* is a slender creeping woody climber about 12 m long, with dark brown branches which contain whitish latex. Its leaves are pinnately compound with entire margins and exude an odourless white gummy substance with slightly acrid taste when cut. It bears numerous flowers and achene (cypsela) fruits. In West Africa, the leaves are used to treat constipation, pain in rheumatism and arthritis, gastroin-

To evaluate the anti-inflammatory effect of the plant, the ethanolic leaf extract (30–300 mg/kg *p.o.*) was examined in the carrageenan-induced foot oedema in chicks and caused a dose-dependent inhibition foot oedema. The highest dose of the

*S. nodiflora* is a common weed usually found growing along the banks of rivers, streams and roadsides of tropical African countries. It is an erect branched annual herb with ascending woody stems branching dichotomously from the base. It leaves occur in opposite pairs, elliptic in shape with finely toothed margins and bear small crowded yellow flowers at nodes. The whole plant is boiled in water and drank for the treatment of convulsion, threatened miscarriage, constipation, arthritis and as

The analgesic effect of the whole plant was investigated in several animal models. The ethanolic extract of the whole plant (100–1000 mg/kg *p.o.*) significantly reduced the number of writhes in mice during an acetic acid-induced writhing assay (ED50 = 141.9 ± 37.16) and inhibited both neurogenic (ED50 = 25.98 ± 14.59) and inflammatory (ED50 = 30.24 ± 18.08) phases of the nociceptive pain produced by formalin *via* adenosinergic mechanisms [87]. In other studies, the hydroethanolic extract of the whole plant (100–1000 mg/kg *p.o.*) caused a significant decrease to pain perception in mechanical, tactile, cold water and thermal hyper-

*T. monadelpha* is an evergreen, small to medium-sized tree with a straight cylindrical bole, smooth greyish outer bark and a pale pink inner wood. Its leaves are alternate, imparipinnately compound. It bears greenish yellow flowers and an obovoid 6-seeded dehiscent capsule. It is commonly known as '*otanduru*' (Akan-Twi) in Ghana and found growing at the river banks near evergreen semi deciduous forests. Various parts of the plant find use in traditional medicine for the treatment of inflammatory condition and neurological disorder such as epilepsy and psychosis [90].

algesia in paclitaxel and vincristine-induced neuropathic pain [88, 89].

**2.20** *Trichilia monadelpha* **(Thonn) JJ De Wilde (***Meliaceae***)**

testinal discomfort, urinary tract and sexually transmitted infection [84].

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

#### *Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants DOI: http://dx.doi.org/10.5772/intechopen.90154*

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

and 89.19 ± 3.81% respectively in the early and late phases [73].

**2.16** *Picralima nitida* **(Stapf ) T. Durand & H. Durand (***Apocynaceae***)**

tory tract infection, constipation and jaundice [76].

suppression of total paw oedema by 82.8 ± 94.6% [78].

**2.17** *Phyllanthus muellerianus* **(Kuntze) Exell. (***Euphorbiaceae***)**

and *κ*-opioid receptors [77, 78].

paralysis and epilepsy [80].

respectively [71].

contralateral limb [75].

writhing with an ED50 of 80.20 ± 0.58 mg/kg and decreased formalin-induced nociception by 83.46 ± 6.67% and 94.56 ± 4.12% in the early and late phases

In other studies, oral administration of the leaf extract (30–300 mg/kg *p.o.*) resulted in a dose-dependent complete reversal of vincristine-induced neuropathic pain in rats [72]. An ecdysteroid called 20-hydroxyecdysone was isolated from the root and was found to inhibit formalin-induced nociception in rats by 71.39 ± 9.19%

The ethanolic root extract (50–400 mg/kg *p.o.*) demonstrated remarkable reduction of carrageenan-induced foot oedema in chicks in both curative (62.52 ± 4.73%) and prophylactic (58.90 ± 11.38%) treatment regimens [74]. Further, the ethanolic leaf extract caused significant reduction in arthritic oedema induced by Freund's adjuvant and prevented the systemic spread of arthritis from the ipsilateral to the

*P. nitida* is a medium sized to large tree which reaches up to 35 m in height with a dense crown, a pale yellow, fine grained inner wood and a cylindrical trunk. The leaves are broadly oblong with hard tiny lateral nerves and bear white flowers with ovoid fruits which turn yellow at maturity. *P. nitida* is widely distributed in the deciduous forests of West and Central Africa. In Ghana, the seeds are locally known as '*akuama*' (Asante-Akan) or '*onwema*' (Fante) and are used for the treatment of pain of various aetiologies as well as fever. Other plant parts find use in folk medicine for the treatment of malaria, fever, worm infestation, venereal disease, respira-

Investigation of the analgesic effects of seeds collected from Ghana established that the aqueous seed extract possessed significant antinociceptive effect in murine models tested by the hot plate assay. Indole alkaloids isolated from the seeds, namely akuammidine, akuammine, akuammicine, akuammigine and pseudoakuammigine also exhibited potent analgesic effects in an isolated tissue and radio-ligand binding assay, demonstrating varying degrees of agonist and antagonist activity at *μ*-, *δ*-,

In anti-inflammatory studies, the hydro-ethanolic extract of the seeds demonstrated a dose-dependent suppression of paw oedema in the carrageenan-induced paw oedema assay. The extract further showed inhibition of chronic inflammation in rat adjuvant-induced arthritis. The total alkaloidal extract at 75–300 mg/kg *p.o.* caused a significant dose-dependent inhibition of total oedema formation in carrageenan-induced paw oedema assay and reduced adjuvant-induced knee joint swelling in rats [79]. Pseudoakuammigine displayed significant dose-dependent

*P. muellerianus* is a straggling shrub about 12 m tall with spreading branches and several short axillary shoots dispersed in the deciduous and secondary forests of tropical Africa. It has simple alternate glabrous leaves and clustered whitish-green flowers. The plant bears fleshy six-seeded smooth capsules which are green when young and black at maturity. The fresh twigs are chewed to prevent toothache and also used to treat dysmenorrhea, dropsy, wound, swelling, oedema, tumour,

The aerial part of the plant was investigated for analgesic and anti-inflammatory

effects in various models. Oral administration of the aqueous extract in doses

**12**

of 30, 100, 300 mg/kg, produced significant antinociceptive effect in the acetic acid-induced abdominal writhing and formalin-induced nociception models in rats [81]. The antinociceptive effect of its major constituent geraniin was demonstrated *via* interaction with opioidergic receptors. Geraniin was found to potentiate the antinociceptive effects of diclofenac and morphine when co-administered [82].

In the carrageenan-induced acute inflammation model, the 70% ethanolic extract of the whole plant and 10 mg/kg of its major constituent geraniin significantly reduced paw oedema by 46.75 ± 4.97% and 61.65 ± 6.70% respectively. The extract and geraniin further attenuated arthritis by reducing total limb swelling in the Freund's adjuvant-induced arthritis model. Histomorphological analysis revealed reduced bone damage in both extract and geraniin treated groups [83].

#### **2.18** *Secamone afzelii* **(Schult.) K. Schum (***Asclepiadaceae***)**

*S. afzelii* is a slender creeping woody climber about 12 m long, with dark brown branches which contain whitish latex. Its leaves are pinnately compound with entire margins and exude an odourless white gummy substance with slightly acrid taste when cut. It bears numerous flowers and achene (cypsela) fruits. In West Africa, the leaves are used to treat constipation, pain in rheumatism and arthritis, gastrointestinal discomfort, urinary tract and sexually transmitted infection [84].

To evaluate the anti-inflammatory effect of the plant, the ethanolic leaf extract (30–300 mg/kg *p.o.*) was examined in the carrageenan-induced foot oedema in chicks and caused a dose-dependent inhibition foot oedema. The highest dose of the extract gave a 44.26% inhibition of oedema [85].

#### **2.19** *Synedrella nodiflora* **(Linn.) Gaertn. (***Asteraceae***)**

*S. nodiflora* is a common weed usually found growing along the banks of rivers, streams and roadsides of tropical African countries. It is an erect branched annual herb with ascending woody stems branching dichotomously from the base. It leaves occur in opposite pairs, elliptic in shape with finely toothed margins and bear small crowded yellow flowers at nodes. The whole plant is boiled in water and drank for the treatment of convulsion, threatened miscarriage, constipation, arthritis and as haemostatic [86].

The analgesic effect of the whole plant was investigated in several animal models. The ethanolic extract of the whole plant (100–1000 mg/kg *p.o.*) significantly reduced the number of writhes in mice during an acetic acid-induced writhing assay (ED50 = 141.9 ± 37.16) and inhibited both neurogenic (ED50 = 25.98 ± 14.59) and inflammatory (ED50 = 30.24 ± 18.08) phases of the nociceptive pain produced by formalin *via* adenosinergic mechanisms [87]. In other studies, the hydroethanolic extract of the whole plant (100–1000 mg/kg *p.o.*) caused a significant decrease to pain perception in mechanical, tactile, cold water and thermal hyperalgesia in paclitaxel and vincristine-induced neuropathic pain [88, 89].

#### **2.20** *Trichilia monadelpha* **(Thonn) JJ De Wilde (***Meliaceae***)**

*T. monadelpha* is an evergreen, small to medium-sized tree with a straight cylindrical bole, smooth greyish outer bark and a pale pink inner wood. Its leaves are alternate, imparipinnately compound. It bears greenish yellow flowers and an obovoid 6-seeded dehiscent capsule. It is commonly known as '*otanduru*' (Akan-Twi) in Ghana and found growing at the river banks near evergreen semi deciduous forests. Various parts of the plant find use in traditional medicine for the treatment of inflammatory condition and neurological disorder such as epilepsy and psychosis [90].

Various solvent extracts (pet-ether, ethyl acetate and methanol) of the stem bark were evaluated for analgesic and anti-inflammatory effect. A significant dosedependent antinociceptive activity in the chemical, thermal and mechanical models of pain was elicited by interaction with opioidergic, muscarinic cholinergic and adenosinergic pathways [91].

The aqueous and pet-ether stem bark extracts suppressed carrageenaninduced foot oedema in chicks by 57.79 ± 3.92% and 63.83 ± 12.0% respectively. In a Complete Freund's Adjuvant-induced arthritis assay, the aqueous extract (100 mg/kg *p.o.*) caused a significant attenuation of chronic inflammation by reducing joint thickness by 64.41 ± 5.56% [92]. Moreover, the stem bark extract caused significant reduction in the high levels of TNF-α, IL-6, malonaldehyde and myeloperoxidase and increased the levels of superoxide dismutase [93] and improved arthritic score by reducing redness, swelling and joint stiffness in rats. Hyperplasia, formation of pannus and exudation of inflammatory cells into synovial spaces were also reduced [94].

#### **2.21** *Vernonia amygdalina* **Delile. (***Compositae***)**

*V. amygdalina* is a widely grown shrub in many African countries including Ghana, Nigeria, Cameroon, Togo, Benin, Guinea and Sierra Leone. It reaches up to about 10 m tall and is severally branched with a greyish-brown smooth bark. Its leaves are ovate-elliptical in shape, simple and alternately arranged with minutely toothed margin. It bears a 10-ribbed achene pubescent dark brown to black fruit. Due to the bitterness of its leaves, the plant is called 'bitter leaf' in many countries. In Ghana, the Akans refer to it as '*awonwene*' (Twi) literally meaning 'bitterness'. The leaves, stem bark and roots are used to treat malaria, fever, worm infestation, skin and nasopharyngeal infection, diarrhoea, dysentery, diabetes and as pain reliever in arthritis and rheumatism [95].

In previous studies, the anti-inflammatory properties of the leaves were evaluated in various models. The ethanol extracts of the young and old leaves (200 mg/kg *p.o.*) caused a significant dose-dependent inhibition of carrageenan-induced cold allodynia, increased the tail withdrawal latency in the tail immersion test and reduced the paw licking time in formalin-induced nociception test in rats *via* opioidergic, nitric oxide cyclic GMP and the muscarinic cholinergic pathways [96].

The young leaf extract at 50, 100 and 200 mg/kg *p.o.* significantly and dosedependently reduced carrageenan-induced foot pad oedema by 59.61%, 67.52% and 86.31% respectively. Similarly, the old leaf extract at same doses exhibited remarkable suppression of oedema formation by 56.11%, 63.37% and 67.41% respectively [96].

#### **2.22** *Wissadula amplissima* **var. rostrata (Schum. & Thonn.) (***Malvaceae***)**

*W. amplissima* is an erect, shrubby herb which grows up to 2.5 m tall on rocky and loamy soils of grassland, bushes and forests in tropical Africa. The leaves have entire or slightly toothed margins, densely pubescent on the lower surface but with sparsely stellate hairs on the dark green upper surface. The leaves are used as a poultice to relief spider bite and sting by venomous insects [25, 97].

The anti-inflammatory activity of the pet ether, chloroform and methanol fractions was investigated in 7-day old chicks and showed significant dose-dependent reduction of carrageenan-induced foot oedema. Maximal oedema inhibition was recorded as 68.25 ± 2.03%, 77.83 ± 0.81% and 62.21 ± 2.61% for the three extracts respectively [98].

**15**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

*X. aethiopica* is a tall evergreen aromatic tree with a smooth greyish-brown bark, severally branched crown and a buttressed bole. Its leaves are coriaceous, green on the upper surface and greenish-brown to orange on the lower surface. It bears small dark brown, cylindrical twisted bean-like aromatic pods, with about 5–8 black seeds per pod. The tree is usually found in lowland rainforests, coastal brackish swamps and deciduous forests of tropical Africa. The fruit is the most important part of the plant and is commonly known as the 'African pepper'. In Ghana it is locally referred to as '*hwentia*' (Twi), '*tso*' (Ewe) and '*soo*' (Ga). It is used as a flavouring in the preparation of soups and for the treatment of inflammatory conditions such as arthritis, bronchitis, rheumatism, lumbago, headache, neuralgia and colic pain [25]. The ethanolic fruits extract and its major diterpene constituent, xylopic acid were investigated for analgesic effects in several pain models. The fruit extract (XAE, 30–300 mg/kg *p.o.*) and xylopic acid (XA, 10–100 mg/kg *p.o.*) inhibited acetic acid-induced visceral nociception, formalin-induced paw pain, thermally-induced as well as carrageenan-induced mechanical and thermal

hyperalgesia [99]. XAE and XA also exhibited anti-hyperalgesic and anti-allodynic properties in vincristine and paclitaxel-induced neuropathic pain [100, 101]. Co-administration of XA and pregabalin synergistically reduced paclitaxel induced neuropathic pain without causing any toxicity [102]. XAE and XA dose-dependently reduced both acute and chronic carrageenan-induced musculoskeletal pain [103] *via* opioidergic, adenosinergic, adrenergic, bradykinin and

In anti-inflammatory studies, the aqueous fruit extract (300 mg/kg *p.o.*) caused a significant reduction of carrageenan-induced paw oedema in mice through inhibition of histamine release from mast cells [105]. Histopathology revealed substantial reduction in mononuclear infiltration, formation of pannus and bone erosion [106]. XA also caused inhibition of inhibition of histamine, serotonin, bradykinin and

*Z. abyssinica* is a thorny, semi-deciduous plant, varying in habit from an erect shrub, a climbing plant or a tree with sagging branches that form a heavy, rounded crown. It usually reaches up to about 12 m tall and has a straight bole. It is commonly known as 'Catch thorn' in English and '*larukluror*' among the Sissala people of Ghana. The root and leaves are useful in folk medicine for treatment of pneumonia, tonsillitis, burn wound, chest pain, migraine and as a general pain-killer [108]. The analgesic and anti-inflammatory effects of the roots were investigated. The hydro-ethanolic root bark extract (30–300 mg/kg, *p.o.*) dose-dependently inhibited acetic acid-, formalin- and glutamate-induced nociception with maximal inhibition of 86.29 ± 2.27%, 84.97 ± 5.35%, and 82.81 ± 5.97% respectively. The paw withdrawal latencies in both tail-immersion and carrageenan-induced hyperalgesia were also prolonged [109]. Moreover, the root extract reversed hyper-nociception induced by intra-plantar injection of TNF-α, IL-1β, bradykinin and prostaglandin E2 *via* interactions with opioidergic, adenosinergic, ATP-sensitive potassium chan-

In an *in-vitro* assay, the hydro-alcoholic root extract at 100 μg/mL inhibited heat and hypotonic-induced haemolysis of human red blood cells by 61.8% and 42.98% respectively. The extracts also inhibited protein (albumin) and bovine serum albumin denaturation. Significant reduction of carrageenan-induced paw oedema

**2.23** *Xylopia aethiopica* **(Dunal) A. Rich. (***Annonaceae***)**

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

prostaglandin nociceptive pathways [104].

prostaglandin E2-induced inflammation [107].

nels and nitric oxide cyclic GMP pathways [110].

**2.24** *Ziziphus abyssinica* **Hochst Ex A. Rich (***Rhamnaceae***)**

### **2.23** *Xylopia aethiopica* **(Dunal) A. Rich. (***Annonaceae***)**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

adenosinergic pathways [91].

synovial spaces were also reduced [94].

**2.21** *Vernonia amygdalina* **Delile. (***Compositae***)**

as pain reliever in arthritis and rheumatism [95].

Various solvent extracts (pet-ether, ethyl acetate and methanol) of the stem bark

were evaluated for analgesic and anti-inflammatory effect. A significant dosedependent antinociceptive activity in the chemical, thermal and mechanical models of pain was elicited by interaction with opioidergic, muscarinic cholinergic and

The aqueous and pet-ether stem bark extracts suppressed carrageenaninduced foot oedema in chicks by 57.79 ± 3.92% and 63.83 ± 12.0% respectively. In a Complete Freund's Adjuvant-induced arthritis assay, the aqueous extract (100 mg/kg *p.o.*) caused a significant attenuation of chronic inflammation by reducing joint thickness by 64.41 ± 5.56% [92]. Moreover, the stem bark extract caused significant reduction in the high levels of TNF-α, IL-6, malonaldehyde and myeloperoxidase and increased the levels of superoxide dismutase [93] and improved arthritic score by reducing redness, swelling and joint stiffness in rats. Hyperplasia, formation of pannus and exudation of inflammatory cells into

*V. amygdalina* is a widely grown shrub in many African countries including Ghana, Nigeria, Cameroon, Togo, Benin, Guinea and Sierra Leone. It reaches up to about 10 m tall and is severally branched with a greyish-brown smooth bark. Its leaves are ovate-elliptical in shape, simple and alternately arranged with minutely toothed margin. It bears a 10-ribbed achene pubescent dark brown to black fruit. Due to the bitterness of its leaves, the plant is called 'bitter leaf' in many countries. In Ghana, the Akans refer to it as '*awonwene*' (Twi) literally meaning 'bitterness'. The leaves, stem bark and roots are used to treat malaria, fever, worm infestation, skin and nasopharyngeal infection, diarrhoea, dysentery, diabetes and

In previous studies, the anti-inflammatory properties of the leaves were evaluated in various models. The ethanol extracts of the young and old leaves (200 mg/kg *p.o.*) caused a significant dose-dependent inhibition of carrageenan-induced cold allodynia, increased the tail withdrawal latency in the tail immersion test and reduced the paw licking time in formalin-induced nociception test in rats *via* opioidergic, nitric oxide cyclic GMP and the muscarinic cholinergic pathways [96]. The young leaf extract at 50, 100 and 200 mg/kg *p.o.* significantly and dosedependently reduced carrageenan-induced foot pad oedema by 59.61%, 67.52% and 86.31% respectively. Similarly, the old leaf extract at same doses exhibited remarkable suppression of oedema formation by 56.11%, 63.37% and 67.41%

**2.22** *Wissadula amplissima* **var. rostrata (Schum. & Thonn.) (***Malvaceae***)**

poultice to relief spider bite and sting by venomous insects [25, 97].

*W. amplissima* is an erect, shrubby herb which grows up to 2.5 m tall on rocky and loamy soils of grassland, bushes and forests in tropical Africa. The leaves have entire or slightly toothed margins, densely pubescent on the lower surface but with sparsely stellate hairs on the dark green upper surface. The leaves are used as a

The anti-inflammatory activity of the pet ether, chloroform and methanol fractions was investigated in 7-day old chicks and showed significant dose-dependent reduction of carrageenan-induced foot oedema. Maximal oedema inhibition was recorded as 68.25 ± 2.03%, 77.83 ± 0.81% and 62.21 ± 2.61% for the three extracts

**14**

respectively [96].

respectively [98].

*X. aethiopica* is a tall evergreen aromatic tree with a smooth greyish-brown bark, severally branched crown and a buttressed bole. Its leaves are coriaceous, green on the upper surface and greenish-brown to orange on the lower surface. It bears small dark brown, cylindrical twisted bean-like aromatic pods, with about 5–8 black seeds per pod. The tree is usually found in lowland rainforests, coastal brackish swamps and deciduous forests of tropical Africa. The fruit is the most important part of the plant and is commonly known as the 'African pepper'. In Ghana it is locally referred to as '*hwentia*' (Twi), '*tso*' (Ewe) and '*soo*' (Ga). It is used as a flavouring in the preparation of soups and for the treatment of inflammatory conditions such as arthritis, bronchitis, rheumatism, lumbago, headache, neuralgia and colic pain [25].

The ethanolic fruits extract and its major diterpene constituent, xylopic acid were investigated for analgesic effects in several pain models. The fruit extract (XAE, 30–300 mg/kg *p.o.*) and xylopic acid (XA, 10–100 mg/kg *p.o.*) inhibited acetic acid-induced visceral nociception, formalin-induced paw pain, thermally-induced as well as carrageenan-induced mechanical and thermal hyperalgesia [99]. XAE and XA also exhibited anti-hyperalgesic and anti-allodynic properties in vincristine and paclitaxel-induced neuropathic pain [100, 101]. Co-administration of XA and pregabalin synergistically reduced paclitaxel induced neuropathic pain without causing any toxicity [102]. XAE and XA dose-dependently reduced both acute and chronic carrageenan-induced musculoskeletal pain [103] *via* opioidergic, adenosinergic, adrenergic, bradykinin and prostaglandin nociceptive pathways [104].

In anti-inflammatory studies, the aqueous fruit extract (300 mg/kg *p.o.*) caused a significant reduction of carrageenan-induced paw oedema in mice through inhibition of histamine release from mast cells [105]. Histopathology revealed substantial reduction in mononuclear infiltration, formation of pannus and bone erosion [106]. XA also caused inhibition of inhibition of histamine, serotonin, bradykinin and prostaglandin E2-induced inflammation [107].

#### **2.24** *Ziziphus abyssinica* **Hochst Ex A. Rich (***Rhamnaceae***)**

*Z. abyssinica* is a thorny, semi-deciduous plant, varying in habit from an erect shrub, a climbing plant or a tree with sagging branches that form a heavy, rounded crown. It usually reaches up to about 12 m tall and has a straight bole. It is commonly known as 'Catch thorn' in English and '*larukluror*' among the Sissala people of Ghana. The root and leaves are useful in folk medicine for treatment of pneumonia, tonsillitis, burn wound, chest pain, migraine and as a general pain-killer [108].

The analgesic and anti-inflammatory effects of the roots were investigated. The hydro-ethanolic root bark extract (30–300 mg/kg, *p.o.*) dose-dependently inhibited acetic acid-, formalin- and glutamate-induced nociception with maximal inhibition of 86.29 ± 2.27%, 84.97 ± 5.35%, and 82.81 ± 5.97% respectively. The paw withdrawal latencies in both tail-immersion and carrageenan-induced hyperalgesia were also prolonged [109]. Moreover, the root extract reversed hyper-nociception induced by intra-plantar injection of TNF-α, IL-1β, bradykinin and prostaglandin E2 *via* interactions with opioidergic, adenosinergic, ATP-sensitive potassium channels and nitric oxide cyclic GMP pathways [110].

In an *in-vitro* assay, the hydro-alcoholic root extract at 100 μg/mL inhibited heat and hypotonic-induced haemolysis of human red blood cells by 61.8% and 42.98% respectively. The extracts also inhibited protein (albumin) and bovine serum albumin denaturation. Significant reduction of carrageenan-induced paw oedema

and a decreased the level of neutrophils in the peritoneal cavity were observed after oral administration of the root extract [111].

## **3. Conclusion**

The Ghanaian flora provides a potent promising source for new therapeutic interventions for local population. The anti-inflammatory and analgesic activities of the crude extracts and fractions of several medicinal plants employed in Ghanaian traditional medicine have been validated in several models. However, the specific bioactive constituents are not yet identified. Therefore further studies to isolate and verify these anti-inflammatory and analgesic compounds are highly recommended. Further evaluation of safety profiles and standardisation of most active plants will add substantial value to the reported bioactivities and make these plants attractive for adaptation to pharmaceutical companies for further development.

### **Acknowledgements**

Authors are grateful to Mr. Yakubu Jibira (Pharmacology Department, KNUST) for assisting with literature retrieval.

## **Conflict of interest**

Authors have no conflict of interest to declare.

## **Author details**

Evelyn Asante-Kwatia\*, Abraham Yeboah Mensah and Michael Frimpong Baidoo Department of Pharmacognosy, Faculty of Pharmacy and Pharmaceutical Sciences, College of Health Sciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana

\*Address all correspondence to: emireku@yahoo.com

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**17**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

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**References**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants DOI: http://dx.doi.org/10.5772/intechopen.90154*

### **References**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

oral administration of the root extract [111].

**3. Conclusion**

**Acknowledgements**

**Conflict of interest**

for assisting with literature retrieval.

Authors have no conflict of interest to declare.

and a decreased the level of neutrophils in the peritoneal cavity were observed after

The Ghanaian flora provides a potent promising source for new therapeutic interventions for local population. The anti-inflammatory and analgesic activities of the crude extracts and fractions of several medicinal plants employed in Ghanaian traditional medicine have been validated in several models. However, the specific bioactive constituents are not yet identified. Therefore further studies to isolate and verify these anti-inflammatory and analgesic compounds are highly recommended. Further evaluation of safety profiles and standardisation of most active plants will add substantial value to the reported bioactivities and make these plants attractive for adaptation to pharmaceutical companies for further development.

Authors are grateful to Mr. Yakubu Jibira (Pharmacology Department, KNUST)

**16**

**Author details**

Kumasi, Ghana

Evelyn Asante-Kwatia\*, Abraham Yeboah Mensah and Michael Frimpong Baidoo Department of Pharmacognosy, Faculty of Pharmacy and Pharmaceutical Sciences, College of Health Sciences, Kwame Nkrumah University of Science and Technology,

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

\*Address all correspondence to: emireku@yahoo.com

provided the original work is properly cited.

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[21] Adeniyi A, Asase A, Ekpe PK, Asitoakor BK, Adu-Gyamfi A, Avekor PY. Ethnobotanical study of medicinal plants from Ghana; confirmation of ethnobotanical uses, and review of biological and toxicological studies on medicinal plants used in Apra Hills Sacred Grove. Journal of Herbal Medicine. 2018;**14**:76-87

[22] Appiah K, Oppong C, Mardani H, Omari R, Kpabitey S, Amoatey C, et al. Medicinal plants used in the Ejisu-Juaben municipality, Southern Ghana: An Ethnobotanical Study. Medicines. 2019;**6**(1):1-27

[23] Asase A, Yohonu DT. Ethnobotanical study of herbal medicines for management of diabetes mellitus in Dangme West District of southern Ghana. Journal of Herbal Medicine. 2016;**6**(4):204-209

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[25] Neuwinger HD. African Traditional Medicine: A Dictionary of Plant Use and Applications. With Supplement: Search System for Diseases. Stuttgart, Germany: Medpharm Scientific; 2000

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[27] Abotsi WKM, Lamptey SB, Boakye-Gyasi E, Woode E. *Albizia zygia* (DC.) JF Macbr. (Leguminosae-Mimosoideae) root extract exhibits anti-nociceptive and antipyretic activities in murine models. Journal of Ethnopharmacology. 2017;**199**:183-193

[28] Oteng-Amoako A, Essien C. *Anopyxis klaineana* (Pierre) Engl. Wageningen, Netherlands: Plant Resources of Tropical Africa/Ressources Végétales de l'Afrique Tropicale; 2011

[29] Mireku EA, Mensah AY, Mensah ML, Tocher DA, Habtemariam S. Anti-inflammatory properties of the stem-bark of *Anopyxis klaineana* and its major constituent. Methyl Angolensate*.* Phytotherapy Research. 2014;**28**(12):1855-1860

[30] Mireku EA, Kusari S, Eckelmann D, Mensah AY, Talontsi FM, Spiteller M.

**19**

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

[38] Danquah CA, Woode E,

inflammatory and antipyretic effects of an ethanolic extract of *Capparis erythrocarpos* Isert roots. Research Journal of Medicinal Plant.

2011;**5**(2):158-168

10.1016/j.jep.2019.111890

Publishers, Gmbh; 2004

Bioresources. 2008;**4**:49-58

2013;**4**(4):301-310

2017;**11**(39):613-620

[44] Mensah AY, Mireku EA, Oppong-Damoah A. Amponsah IK. Anti-inflammatory and antioxidant activities of *Commelina diffusa*

[45] Adu-Amoah L, Agyare C, Kisseih E, Ayande PG, Mensah KB. Toxicity assessment of *Erythrophleum ivorense* and *Parquetina nigrescens*. Toxicology Reports. 2014;**1**:411-440

(Commelinaceae). 2014;**2**(10):1159-1165

Gyasi EB, Duwiejua M, Ansah C. Anti-

[39] Twumasi M, Tandoh A, Mante P, Ekuadzi E, Boakye-Gyasi M, Benneh C, et al. Leaves and stems of *Capparis erythrocarpos*, more sustainable than roots, show antiarthritic effects. Journal of Ethnopharmacology. 2019;**238**. DOI:

[40] Arbonnier M. Trees, Shrubs and Lianas of West African Dry Zones. Paris, France: CIRAD-MNHN, Margraf

[41] Duwiejua M, Panyin AB, Weremfo A, Woode E, Ansah C. Antinociceptive activity of the ethanolic extract of the root bark of *Cassia sieberiana* (Fam. Caesalpinaceae). Journal of Pharmacy &

[42] Donkor K, Okine LN, Abotsi WK, Woode E. Antiinflammatory and anti-nociceptive effects of ethyl acetate fraction of root bark of *Cassia sieberiana* DC in murine models. Pharmacologia.

[43] Donkor K, Woode E, Okine LK. Immunoregulatory activity of root bark of *Cassia sieberiana* DC in a modified adjuvant-induced arthritis in rat. Journal of Medicinal Plants Research.

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

Anti-inflammatory tirucallane triterpenoids from *Anopyxis klaineana* Pierre (Engl.),(Rhizophoraceae). Fitoterapia. 2015;**106**:84-91

[31] Murti Y, Yogi B, Pathak D. Pharmacognostic standardization of leaves of *Calotropis procera* (Ait.) R. Br. (Asclepiadaceae). International Journal of Ayurveda Research. 2010;**1**(1):14-17

[32] Yogi B, Gupta SK, Mishra A.

2016;**5**:74-81

*Calotropis procera* (Madar): A medicinal plant of various therapeutic uses—A review. Bulletin of Environment, Pharmacology and Life Sciences.

[33] Arbonnier M. Arbres, arbustes et lianes des zones sèches d'Afrique de l'Ouest. Paris, France: CIRAD–MNHN, Margraf publishers, Gmbh; 2002

[34] Obese E, Ameyaw EO, Biney RP, Henneh IT, Edzeamey FJ, Woode E. Phytochemical screening and antiinflammatory properties of the

hydroethanolic leaf extract of *Calotropis procera* (Ait). R. Br.(Apocynaceae). Journal of Pharmaceutical Research International. 2018;**23**(1):1-11

[35] Twumasi MA, Ekuadzi E, Mante PK, Boakye-Gyasi ME, Mensah MLK, Woode E. Pharmacognostic studies of the leaves, stem and root of *Capparis erythrocarpos* Isert

(Capparaceae). Pharmacognosy Journal.

[36] Woode E, Ansah C. Antinociceptive

Brew-Daniels H, Appiah AA, Ocloo A. *In-vivo* comparative anti-inflammatory and analgesic activities of root bark, stem and leaf extracts of *Capparis erythrocarpus* (Capparaceae). Pharmacognosy Journal.

effect of an Ethanolic extract of *Capparis erythrocarpos* Isert root in the mice formalin test. International Journal of Pharmacology. 2009;**5**(6):354-361

[37] Kumatia EK, Antwi S,

2019;**11**(3):515-520

2019;**11**(1):112-118

#### *Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants DOI: http://dx.doi.org/10.5772/intechopen.90154*

Anti-inflammatory tirucallane triterpenoids from *Anopyxis klaineana* Pierre (Engl.),(Rhizophoraceae). Fitoterapia. 2015;**106**:84-91

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

[23] Asase A, Yohonu DT. Ethnobotanical study of herbal

medicines for management of diabetes mellitus in Dangme West District of southern Ghana. Journal of Herbal Medicine. 2016;**6**(4):204-209

[24] Sengupta R, Sheorey SD, Hinge MA. Analgesic and anti-inflammatory plants: An updated review.

International Journal of Pharmaceutical

[25] Neuwinger HD. African Traditional Medicine: A Dictionary of Plant Use and Applications. With Supplement: Search System for Diseases. Stuttgart, Germany: Medpharm Scientific; 2000

Sciences Review and Research.

[26] Abotsi WKM, Lamptey SB, Afrane S, Boakye-Gyasi E, Umoh RU, Woode E. An evaluation of the antiinflammatory, antipyretic and analgesic

effects of hydroethanol leaf extract of *Albizia zygia* in animal models. Pharmaceutical Biology.

[27] Abotsi WKM, Lamptey SB, Boakye-Gyasi E, Woode E. *Albizia zygia* (DC.) JF Macbr. (Leguminosae-Mimosoideae) root extract exhibits anti-nociceptive and antipyretic activities in murine models. Journal of Ethnopharmacology. 2017;**199**:183-193

[28] Oteng-Amoako A, Essien C. *Anopyxis klaineana* (Pierre) Engl. Wageningen, Netherlands: Plant

Resources of Tropical Africa/Ressources Végétales de l'Afrique Tropicale; 2011

[30] Mireku EA, Kusari S, Eckelmann D, Mensah AY, Talontsi FM, Spiteller M.

[29] Mireku EA, Mensah AY, Mensah ML, Tocher DA, Habtemariam S. Anti-inflammatory properties of the stem-bark of *Anopyxis klaineana* and its major constituent. Methyl Angolensate*.* Phytotherapy Research.

2014;**28**(12):1855-1860

2017;**55**(1):338-348

2012;**12**(2):114-119

[16] Patel M, Murugananthan G, Gowda K. *In-vivo* animal models in preclinical evaluation of antiinflammatory activity—A review. International Journalof Pharmaceutical

Research and Allied Sciences.

[17] Hariram Nile S, Won PS. Optimized methods for *in-vitro* and *in-vivo* antiinflammatory assays and its applications in herbal and synthetic drug analysis. Mini Reviews in Medicinal Chemistry.

[18] Morris CJ. Carrageenan-induced paw edema in the rat and mouse. In: Winyard PG, Willoughby DA, editors. Inflammation Protocols. Totowa, NJ: Humana Press; 2003. pp. 115-121

[19] Patel PK, Sahu J, Chandel SS. A detailed review on nociceptive models for the screening of analgesic activity in experimental animals. International Journal of Neurologic Physical Therapy.

[20] Agyei-Baffour P, Kudolo A, Quansah DY, Boateng D. Integrating herbal medicine into mainstream healthcare in Ghana: Clients'

acceptability, perceptions and disclosure of use. BMC Complementary and Alternative Medicine. 2017;**17**(513):1-9

[21] Adeniyi A, Asase A, Ekpe PK, Asitoakor BK, Adu-Gyamfi A, Avekor PY. Ethnobotanical study of medicinal plants from Ghana; confirmation of ethnobotanical uses, and review of biological and toxicological studies on medicinal plants used in Apra Hills Sacred Grove. Journal of Herbal Medicine.

[22] Appiah K, Oppong C, Mardani H, Omari R, Kpabitey S, Amoatey C, et al. Medicinal plants used in the Ejisu-Juaben municipality, Southern Ghana: An Ethnobotanical Study. Medicines.

2012;**1**(2):1-5

2013;**13**(1):95-100

2016;**2**:44-50

2018;**14**:76-87

2019;**6**(1):1-27

**18**

[31] Murti Y, Yogi B, Pathak D. Pharmacognostic standardization of leaves of *Calotropis procera* (Ait.) R. Br. (Asclepiadaceae). International Journal of Ayurveda Research. 2010;**1**(1):14-17

[32] Yogi B, Gupta SK, Mishra A. *Calotropis procera* (Madar): A medicinal plant of various therapeutic uses—A review. Bulletin of Environment, Pharmacology and Life Sciences. 2016;**5**:74-81

[33] Arbonnier M. Arbres, arbustes et lianes des zones sèches d'Afrique de l'Ouest. Paris, France: CIRAD–MNHN, Margraf publishers, Gmbh; 2002

[34] Obese E, Ameyaw EO, Biney RP, Henneh IT, Edzeamey FJ, Woode E. Phytochemical screening and antiinflammatory properties of the hydroethanolic leaf extract of *Calotropis procera* (Ait). R. Br.(Apocynaceae). Journal of Pharmaceutical Research International. 2018;**23**(1):1-11

[35] Twumasi MA, Ekuadzi E, Mante PK, Boakye-Gyasi ME, Mensah MLK, Woode E. Pharmacognostic studies of the leaves, stem and root of *Capparis erythrocarpos* Isert (Capparaceae). Pharmacognosy Journal. 2019;**11**(1):112-118

[36] Woode E, Ansah C. Antinociceptive effect of an Ethanolic extract of *Capparis erythrocarpos* Isert root in the mice formalin test. International Journal of Pharmacology. 2009;**5**(6):354-361

[37] Kumatia EK, Antwi S, Brew-Daniels H, Appiah AA, Ocloo A. *In-vivo* comparative anti-inflammatory and analgesic activities of root bark, stem and leaf extracts of *Capparis erythrocarpus* (Capparaceae). Pharmacognosy Journal. 2019;**11**(3):515-520

[38] Danquah CA, Woode E, Gyasi EB, Duwiejua M, Ansah C. Antiinflammatory and antipyretic effects of an ethanolic extract of *Capparis erythrocarpos* Isert roots. Research Journal of Medicinal Plant. 2011;**5**(2):158-168

[39] Twumasi M, Tandoh A, Mante P, Ekuadzi E, Boakye-Gyasi M, Benneh C, et al. Leaves and stems of *Capparis erythrocarpos*, more sustainable than roots, show antiarthritic effects. Journal of Ethnopharmacology. 2019;**238**. DOI: 10.1016/j.jep.2019.111890

[40] Arbonnier M. Trees, Shrubs and Lianas of West African Dry Zones. Paris, France: CIRAD-MNHN, Margraf Publishers, Gmbh; 2004

[41] Duwiejua M, Panyin AB, Weremfo A, Woode E, Ansah C. Antinociceptive activity of the ethanolic extract of the root bark of *Cassia sieberiana* (Fam. Caesalpinaceae). Journal of Pharmacy & Bioresources. 2008;**4**:49-58

[42] Donkor K, Okine LN, Abotsi WK, Woode E. Antiinflammatory and anti-nociceptive effects of ethyl acetate fraction of root bark of *Cassia sieberiana* DC in murine models. Pharmacologia. 2013;**4**(4):301-310

[43] Donkor K, Woode E, Okine LK. Immunoregulatory activity of root bark of *Cassia sieberiana* DC in a modified adjuvant-induced arthritis in rat. Journal of Medicinal Plants Research. 2017;**11**(39):613-620

[44] Mensah AY, Mireku EA, Oppong-Damoah A. Amponsah IK. Anti-inflammatory and antioxidant activities of *Commelina diffusa* (Commelinaceae). 2014;**2**(10):1159-1165

[45] Adu-Amoah L, Agyare C, Kisseih E, Ayande PG, Mensah KB. Toxicity assessment of *Erythrophleum ivorense* and *Parquetina nigrescens*. Toxicology Reports. 2014;**1**:411-440

[46] Armah FA, Annan K, Mensah AY, Amponsah IK, Tocher DA, Habtemariam S. Erythroivorensin: A novel anti-inflammatory diterpene from the root-bark of *Erythrophleum ivorense* (A Chev.). Fitoterapia. 2015;**105**:37-42

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Pharmacognostic standardisation of *Hilleria latifolia* (Lam.) H. Walt. (Phytolaccaceae). Asian Pacific Journal of Tropical Biomedicine. 2014;**4**(12):941-946

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

the stem bark of *Ficus exasperata* Vahl (Moraceae). Journal of Scientific and Innovative Research. 2013;**2**(5):880-887

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[54] Dickson R, Ekuadzi E, Annan K, Komlaga G. Antibacterial, antiinflammatory, and antioxidant effects of the leaves and stem bark of *Glyphaea brevis* (Spreng) Monachino (Tiliaceae): A comparative study. Pharmacognosy

[55] Obiri DD, Osafo N, Abotsi RE. Antiallergic and antiarthritic effects of stem bark extract of *Glyphaea brevis* (Spreng) Monachino (Tiliaceae) in murine models. ISRN Pharmacology.

[56] Asase A, Oteng-Yeboah AA, Odamtten GT, Simmonds MS. Ethnobotanical study of some Ghanaian anti-malarial plants. Journal of

[57] Ameyaw EO, Kukuia KKE, Thomford AK, Kyei S, Mante PK, Boampong JN. Analgesic properties of aqueous leaf extract of *Haematostaphis barteri*: Involvement of ATP-sensitive potassium channels, adrenergic, opioidergic, muscarinic, adenosinergic and serotoninergic pathways. Journal of Basic and Clinical Physiology and Pharmacology. 2016;**27**(6):557-561

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Mensah AY, Amponsah IK, Tocher DA, Habtemariam S. Erythroivorensin: A novel anti-inflammatory diterpene from the root-bark of *Erythrophleum ivorense* (A Chev.). Fitoterapia. 2015;**105**:37-42

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Africa/ Ressources végétales de l'Afrique tropicale. 2010. Available from: http:// www.prota4u.org/search.asp. [Accessed:

**20**

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[75] Woode E, Boakye-Gyasi E, Danquah C, Ansah C, Duwiejua M. Anti-arthritic effects of *Palisota hirsuta* K. Schum. leaf extract in Freund's adjuvant-induced arthritis in rats. International Journal of Pharmacology. 2009;**5**:181-190

[76] Erharuyi O, Falodun A, Langer P. Medicinal uses, phytochemistry and pharmacology of *Picralima nitida* (Apocynaceae) in tropical diseases: A review. Asian Pacific Journal of Tropical Medicine. 2014;**7**(1):1-8

[77] Menzies JR, Paterson SJ, Duwiejua M, Corbett AD. Opioid activity of alkaloids extracted from *Picralima nitida* (fam. Apocynaceae). European Journal of Pharmacology. 1998;**350**(1):101-108

[78] Duwiejua M, Woode E, Obiri D. Pseudo-akuammigine, an alkaloid from *Picralima nitida* seeds, has antiinflammatory and analgesic actions in rats. Journal of Ethnopharmacology. 2002;**81**(1):73-79

[79] Woode E, Obiri D, Ansah C, Duwiejua M, Kuffuor G. Total alkaloidal extract of *Picralima nitida* (Fam. Apocynaceae) seeds has antiinflammatory actions. Journal of the Ghana Science Association. 2006;**8**(1):70-78

[80] Ben-Bala K. *Phyllanthus muellerianus (Kuntze) Exell*. Plant Resources of Tropical Africa/Ressources Végétales de L'Afrique Tropicale. 2008. Available

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[82] Boakye-Gyasi E, Kasanga EA, Ameyaw EO, Abotsi WKM, Biney RP, Agyare C, et al. An isobolographic analysis of the anti-nociceptive effect of geraniin in combination with morphine or diclofenac. Journal of Basic and Clinical Physiology and Pharmacology. 2018;**29**(2):201-209

[83] Boakye YD, Agyare C, Abotsi WKM, Ayande PG, Ossei PPS. Antiinflammatory activity of aqueous leaf extract of *Phyllanthus muellerianus* (Kuntze) Exell. and its major constituent, geraniin. Journal of Ethnopharmacology. 2016;**187**:17-27

[84] Abere TA, Onwukaeme DN. Pharmacognostic evaluation of the leaves of *Secamone afzelii* (Schult) K Schum (Asclepiadaceae). Tropical Journal of Pharmaceutical Research. 2012;**11**(1):125-131

[85] Mensah A, Mireku E, Okwuonu V. Anti-inflammatory and anti-oxidant activities of *Secamone afzelii* (Rhoem) Ascleipiadaceae. Journal of Medical and Biomedical Sciences. 2014;**3**(1):23-30

[86] Bhogaonkar P, Dagawal M, Ghorpade D. Pharmacognostic studies and antimicrobial activity of *Synedrella nodiflora* (L.) Gaertn. Bioscience Discovery. 2011;**2**(3):317-321

[87] Woode E, Amoateng P, Ansah C, Duwiejua M. Anti-nociceptive effects of an ethanolic extract of the whole plant of *Synedrella nodiflora* (L.)

**23**

2016;**7**(1):32-43

*Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants*

[94] Ben IO, Woode E, Koffuor GA, Boakye-Gyasi E, Titiloye NA. Effect of *Trichilia monadelpha* (Meliaceae) extracts on bone histomorphology in complete Freund's adjuvant-induced arthritis. Journal of Intercultural Ethnopharmacology. 2017;**6**(2):177

[95] Ucheck FF. *Vernonia Amygdalina Delile*. Plant Resources of Tropical Africa/Ressources végétales de l'Afrique Tropicale. 2004. Available from: http:// www.prota4u.org/search.asp. [Accessed:

Acheampong DO, Kyei F, Adokoh CK, Ofori EG, et al. Anti-inflammatory, anti-nociceptive and antipyretic activity of young and old leaves of *Vernonia amygdalina*. Biomedicine & Pharmacotherapy. 2019;**111**:1187-1203

[97] Oyen LPA. Wissadula rostrata (Schumach. & Thonn.) Planchon ex Hook. f. Plant Resources of Tropical Africa/Ressources Végétales de l'Afrique Tropicale. 2011. Available from: http:// www.prota4u.org/search.asp. [Accessed:

[98] Mensah A, Donkor P, Fleischer T. Anti-inflammatory and antioxidant activities of the leaves of *Wissadula amplissima* var Rostrata. African Journal of Traditional, Complementary

Boakye-Gyasi E, Abotsi WK. Analgesic effects of an ethanol extract of the fruits of *Xylopia aethiopica* (Dunal) A. Rich (Annonaceae) and the major constituent, xylopic acid in murine models. Journal of Pharmacy & Bioallied Sciences. 2012;**4**(4):291

and Alternative Medicines.

[99] Woode E, Ameyaw EO,

[100] Ameyaw EO, Woode E, Boakye-Gyasi E, Abotsi WK, Kyekyeku JO, Adosraku RK. Antiallodynic and anti-hyperalgesic effects

01 August 2019]

01 August 2019]

2011;**8**(2):185-195

[96] Asante D-B, Henneh IT,

*DOI: http://dx.doi.org/10.5772/intechopen.90154*

[88] Amoateng P, Adjei S, Osei-Safo D, Ameyaw EO, Ahedor B, N'guessan BB, et al. A hydro-ethanolic extract of *Synedrella nodiflora* (L.) Gaertn ameliorates hyperalgesia and allodynia in vincristine-induced neuropathic pain in rats. Journal of Basic and Clinical Physiology and Pharmacology.

[89] Amoateng P, Adjei S, Osei-Safo D, Kukuia KKE, Kretchy IA, Sarkodie JA, et al. Analgesic effects of a hydroethanolic whole plant extract of *Synedrella nodiflora* (L.) Gaertn in paclitaxel-induced neuropathic pain in rats. BMC Research Notes.

[90] Lemmens R. *Trichilia monadelpha* (Thonn.) JJ de Wilde. Plant Resources of Tropical Africa. 2008;**7**(1):561-563

[91] Woode E, Amoh-Barimah AK, Abotsi WKM, Ainooson GK, Owusu G. Analgesic effects of stem bark extracts of *Trichilia monadelpha* (Thonn.) JJ De Wilde. Indian Journal of Pharmacology. 2012;**44**(6):765-773

[92] Ainooson G, Owusu G, Woode E, Ansah C, Annan K. *Trichilia monadelpha* bark extracts inhibit carrageenaninduced foot-oedema in the 7-day old chick and the oedema associated with adjuvant-induced arthritis in rats. African Journal of Traditional, Complementary and Alternative Medicines. 2012;**9**(1):8-16

[93] Ben IO, Woode E, Koffuor GA, Boakye-Gyasi E, Ehigiator BE. Effect of *Trichilia monadelpha* (Thonn.) JJ De Wilde (Meliaceae) extracts on haematology, cytokines and oxidative stress biomarkers in rats adjuvantinduced arthritis. Pharmacologia.

Gaertn in mice: Involvement of adenosinergic mechanisms. Journal of Pharmacology and Toxicology.

2009;**4**(1):17-29

2015;**26**(4):383-394

2017;**10**(226):1-7

#### *Analgesic and Anti-Inflammatory Effect of Ghanaian Medicinal Plants DOI: http://dx.doi.org/10.5772/intechopen.90154*

Gaertn in mice: Involvement of adenosinergic mechanisms. Journal of Pharmacology and Toxicology. 2009;**4**(1):17-29

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

from: http://www.prota4u.org/search. asp. [Accessed: 01 August 2019]

[81] Boakye-Gyasi E, Kasange EA, Biney RP, Boadu-Mensah K, Agyare C, Woode E. Anti-nociceptive effects of geraniin and an aqueous extract of the aerial parts of *Phyllanthus muellerianus* (Kuntze) Exell. in murine models of chemical nociception. Iranian Journal of Pharmaceutical Sciences.

[82] Boakye-Gyasi E, Kasanga EA, Ameyaw EO, Abotsi WKM, Biney RP, Agyare C, et al. An isobolographic analysis of the anti-nociceptive effect of geraniin in combination with morphine or diclofenac. Journal of Basic and Clinical Physiology and Pharmacology.

[83] Boakye YD, Agyare C, Abotsi WKM,

inflammatory activity of aqueous leaf extract of *Phyllanthus muellerianus* (Kuntze) Exell. and its major constituent, geraniin. Journal of Ethnopharmacology. 2016;**187**:17-27

2016;**12**(3):17-30

2018;**29**(2):201-209

2012;**11**(1):125-131

Ayande PG, Ossei PPS. Anti-

[84] Abere TA, Onwukaeme DN. Pharmacognostic evaluation of the leaves of *Secamone afzelii* (Schult) K Schum (Asclepiadaceae). Tropical Journal of Pharmaceutical Research.

[85] Mensah A, Mireku E, Okwuonu V. Anti-inflammatory and anti-oxidant activities of *Secamone afzelii* (Rhoem) Ascleipiadaceae. Journal of Medical and Biomedical Sciences. 2014;**3**(1):23-30

[86] Bhogaonkar P, Dagawal M, Ghorpade D. Pharmacognostic studies and antimicrobial activity of *Synedrella nodiflora* (L.) Gaertn. Bioscience Discovery. 2011;**2**(3):317-321

[87] Woode E, Amoateng P, Ansah C, Duwiejua M. Anti-nociceptive effects of an ethanolic extract of the whole plant of *Synedrella nodiflora* (L.)

K. Schum (Commelinaceae). Journal of Applied Pharmaceutical Science.

[74] Boakye-Gyasi E, Woode E, Ainooson G, Obiri D, Ansah C, Duwejua M, et al. Anti-inflammatory and antipyretic effects of an ethanolic extract of *Palisota hirsuta* K. Schum roots. African Journal of Pharmacy and Pharmacology. 2008;**2**(9):191-199

[75] Woode E, Boakye-Gyasi E, Danquah C, Ansah C, Duwiejua M. Anti-arthritic effects of *Palisota hirsuta* K. Schum. leaf extract in Freund's adjuvant-induced arthritis in rats. International Journal of Pharmacology.

[76] Erharuyi O, Falodun A, Langer P. Medicinal uses, phytochemistry and pharmacology of *Picralima nitida* (Apocynaceae) in tropical diseases: A review. Asian Pacific Journal of Tropical

Duwiejua M, Corbett AD. Opioid activity of alkaloids extracted from *Picralima* 

European Journal of Pharmacology.

[78] Duwiejua M, Woode E, Obiri D. Pseudo-akuammigine, an alkaloid from *Picralima nitida* seeds, has antiinflammatory and analgesic actions in rats. Journal of Ethnopharmacology.

[79] Woode E, Obiri D, Ansah C,

extract of *Picralima nitida* (Fam. Apocynaceae) seeds has antiinflammatory actions. Journal of the Ghana Science Association.

Duwiejua M, Kuffuor G. Total alkaloidal

[80] Ben-Bala K. *Phyllanthus muellerianus (Kuntze) Exell*. Plant Resources of Tropical Africa/Ressources Végétales de L'Afrique Tropicale. 2008. Available

2016;**6**(10):147-153

2009;**5**:181-190

Medicine. 2014;**7**(1):1-8

[77] Menzies JR, Paterson SJ,

*nitida* (fam. Apocynaceae).

1998;**350**(1):101-108

2002;**81**(1):73-79

2006;**8**(1):70-78

**22**

[88] Amoateng P, Adjei S, Osei-Safo D, Ameyaw EO, Ahedor B, N'guessan BB, et al. A hydro-ethanolic extract of *Synedrella nodiflora* (L.) Gaertn ameliorates hyperalgesia and allodynia in vincristine-induced neuropathic pain in rats. Journal of Basic and Clinical Physiology and Pharmacology. 2015;**26**(4):383-394

[89] Amoateng P, Adjei S, Osei-Safo D, Kukuia KKE, Kretchy IA, Sarkodie JA, et al. Analgesic effects of a hydroethanolic whole plant extract of *Synedrella nodiflora* (L.) Gaertn in paclitaxel-induced neuropathic pain in rats. BMC Research Notes. 2017;**10**(226):1-7

[90] Lemmens R. *Trichilia monadelpha* (Thonn.) JJ de Wilde. Plant Resources of Tropical Africa. 2008;**7**(1):561-563

[91] Woode E, Amoh-Barimah AK, Abotsi WKM, Ainooson GK, Owusu G. Analgesic effects of stem bark extracts of *Trichilia monadelpha* (Thonn.) JJ De Wilde. Indian Journal of Pharmacology. 2012;**44**(6):765-773

[92] Ainooson G, Owusu G, Woode E, Ansah C, Annan K. *Trichilia monadelpha* bark extracts inhibit carrageenaninduced foot-oedema in the 7-day old chick and the oedema associated with adjuvant-induced arthritis in rats. African Journal of Traditional, Complementary and Alternative Medicines. 2012;**9**(1):8-16

[93] Ben IO, Woode E, Koffuor GA, Boakye-Gyasi E, Ehigiator BE. Effect of *Trichilia monadelpha* (Thonn.) JJ De Wilde (Meliaceae) extracts on haematology, cytokines and oxidative stress biomarkers in rats adjuvantinduced arthritis. Pharmacologia. 2016;**7**(1):32-43

[94] Ben IO, Woode E, Koffuor GA, Boakye-Gyasi E, Titiloye NA. Effect of *Trichilia monadelpha* (Meliaceae) extracts on bone histomorphology in complete Freund's adjuvant-induced arthritis. Journal of Intercultural Ethnopharmacology. 2017;**6**(2):177

[95] Ucheck FF. *Vernonia Amygdalina Delile*. Plant Resources of Tropical Africa/Ressources végétales de l'Afrique Tropicale. 2004. Available from: http:// www.prota4u.org/search.asp. [Accessed: 01 August 2019]

[96] Asante D-B, Henneh IT, Acheampong DO, Kyei F, Adokoh CK, Ofori EG, et al. Anti-inflammatory, anti-nociceptive and antipyretic activity of young and old leaves of *Vernonia amygdalina*. Biomedicine & Pharmacotherapy. 2019;**111**:1187-1203

[97] Oyen LPA. Wissadula rostrata (Schumach. & Thonn.) Planchon ex Hook. f. Plant Resources of Tropical Africa/Ressources Végétales de l'Afrique Tropicale. 2011. Available from: http:// www.prota4u.org/search.asp. [Accessed: 01 August 2019]

[98] Mensah A, Donkor P, Fleischer T. Anti-inflammatory and antioxidant activities of the leaves of *Wissadula amplissima* var Rostrata. African Journal of Traditional, Complementary and Alternative Medicines. 2011;**8**(2):185-195

[99] Woode E, Ameyaw EO, Boakye-Gyasi E, Abotsi WK. Analgesic effects of an ethanol extract of the fruits of *Xylopia aethiopica* (Dunal) A. Rich (Annonaceae) and the major constituent, xylopic acid in murine models. Journal of Pharmacy & Bioallied Sciences. 2012;**4**(4):291

[100] Ameyaw EO, Woode E, Boakye-Gyasi E, Abotsi WK, Kyekyeku JO, Adosraku RK. Antiallodynic and anti-hyperalgesic effects of an ethanolic extract and xylopic acid from the fruits of *Xylopia aethiopica* in murine models of neuropathic pain. Pharmacognosy Research. 2014;**6**(2):172-179

[101] Ameyaw E, Boampong J, Kukuia K, Amoateng P, Obese E, Osei-Sarpong C, et al. Effect of xylopic acid on paclitaxel-induced neuropathic pain in rats. Journal of Medical and Biomedical Sciences. 2013;**2**(4):6-12

[102] Ofori AE, Eric W, Samuel K, Robert BP, Nyarko BJ. Anti-nociceptive synergism of pregabalin and xylopic acid co-administration in paclitaxel induced neuropathy: Isobolographic analysis. Pharmacognosy Journal. 2015;**7**(6):363-368

[103] Woode E, Ameyaw EO, Boakye-Gyasi E, Abotsi WKM, Oppong Kyekyeku J, Adosraku R, et al. Effects of an ethanol extract and the diterpene, xylopic acid, of *Xylopia aethiopica* fruits in murine models of musculoskeletal pain. Pharmaceutical Biology. 2016;**54**(12):2978-2986

[104] Woode E, Ameyaw EO, Ainooson G, Abotsi W, Gyasi E, Kyekyeku J. Analgesic effects of an ethanol extract of the fruits of *Xylopia aethiopica* and xylopic acid in murine models of pain: Possible mechanism(s). Pharmacologia. 2013;**4**(4):285-300

[105] Obiri DD, Osafo N. Aqueous ethanol extract of the fruit of *Xylopia aethiopica* (Annonaceae) exhibits anti-anaphylactic and anti-inflammatory actions in mice. Journal of Ethnopharmacology. 2013;**148**(3):940-945

[106] Obiri DD, Osafo N, Ayande PG, Antwi AO. *Xylopia aethiopica* (Annonaceae) fruit extract suppresses Freund's adjuvant-induced arthritis in Sprague-Dawley rats. Journal of Ethnopharmacology. 2014;**152**(3):522-531

[107] Osafo N, Obiri DD, Antwi AO, Yeboah OK. The acute anti-inflammatory action of xylopic acid isolated from *Xylopia aethiopica*. Journal of Basic and Clinical Physiology and Pharmacology. 2018;**29**(6):659-669

[108] Ruffo CK, Birnie A, Tengnäs B. Edible Wild Plants of Tanzania. Vol. 27. Nairobi, Kenya: Regional Land Management Unit, RELMA/Sida; 2002

[109] Boakye-Gyasi E, Henneh IT, Abotsi WKM, Ameyaw EO, Woode E. Hydro-ethanolic leaf extract of *Ziziphus abyssinica* Hochst Ex A. Rich (Rhamnaceae) exhibits anti-nociceptive effects in murine models. BMC Complementary and Alternative Medicine. 2017;**17**(231):1-12

[110] Boakye-Gyasi E, Henneh IT, Abotsi WKM, Ameyaw EO, Woode E. Possible mechanisms involved in the anti-nociceptive effects of hydroethanolic leaf extract of *Ziziphus abyssinica*. Pharmaceutical Biology. 2017;**55**(1):1962-1971

[111] Henneh IT, Ameyaw EO, Biney RP, Armah FA, Obese E, Konjah D, et al. *Ziziphus abyssinica* hydro-ethanolic root bark extract attenuates acute inflammation possibly through membrane stabilization and inhibition of protein denaturation and neutrophil degranulation. West African Journal of Pharmacy. 2018;**29**(2):81-94

**25**

**Chapter 2**

**Abstract**

resistance gene

**1. Introduction**

Antimicrobial Potential of Genes

With the advancements in agriculture, farming community less or more started to rely on synthetic chemicals to increase the crop production and protection. But the negative impact of these chemicals on environment and cropping system urges the scientists to discover some new ways to combat with crop disease. By keeping in view, garlic is a well-known economically important vegetable throughout the world and recognized as reservoir for a number of bioactive compounds to treat various diseases; scientists have developed a strategy to identify and isolate antimicrobial genes from garlic. By using *B. subtilis* expression systems, a total of 48 antimicrobial genes, including *AsR 416*, were identified with the potential to significantly retard the growth of economically important fungal and bacterial pathogens. Furthermore, these antimicrobial genes exhibited the thermal stability along with nontoxic effects on mammalian blood cells, which indicate its potential use in the development of human medicines. These genes can revolutionize the way to treat with pathogens and also give a new wave of knowledge to explore the other organisms for the search of antimicrobial genes. This will also help to search the

from Garlic (*Allium sativum* L.)

*Hafiz Muhammad Khalid Abbas, Xi Kong, Jia Wu,* 

other cost-effective ways for the treatment of plant and human diseases.

Garlic (*Allium sativum* L.) is one of the most important species of the genus *Allium* and recognized as economically important vegetable throughout the world, especially around the Mediterranean basin where it is considered as main agricultural product [1, 2]. It is also of great importance because of its therapeutic properties and health-related benefits against various kinds of diseases such as aches, deafness, diarrhea, constipation, tumors, and respiratory problems. Health benefits from *Allium* species, especially garlic, have been used for centuries to treat various kinds of disorders, and still, there is need of research to explore its healthrelated potential [3–5]. It is a historic medicinal plant, originated from central Asia about 6000 years ago, and had been started to use as medicine in India since 5000 years ago and 3000 years ago in China [6, 7]. Volatile sulfur compounds, especially thiosulfates, responsible for pungent aroma, are the main compounds

**Keywords:** *Allium sativum* L., antimicrobial peptide, *Bacillus subtilis*,

*Mohsin Ali and Wubei Dong*

### **Chapter 2**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

[107] Osafo N, Obiri DD, Antwi AO, Yeboah OK. The acute anti-inflammatory action of xylopic acid isolated from *Xylopia aethiopica*. Journal of Basic and Clinical Physiology and Pharmacology.

[108] Ruffo CK, Birnie A, Tengnäs B. Edible Wild Plants of Tanzania. Vol. 27. Nairobi, Kenya: Regional Land Management Unit, RELMA/Sida; 2002

[109] Boakye-Gyasi E, Henneh IT, Abotsi WKM, Ameyaw EO, Woode E. Hydro-ethanolic leaf extract of *Ziziphus* 

(Rhamnaceae) exhibits anti-nociceptive

[111] Henneh IT, Ameyaw EO, Biney RP, Armah FA, Obese E, Konjah D, et al. *Ziziphus abyssinica* hydro-ethanolic root bark extract attenuates acute inflammation possibly through

membrane stabilization and inhibition of protein denaturation and neutrophil degranulation. West African Journal of

Pharmacy. 2018;**29**(2):81-94

*abyssinica* Hochst Ex A. Rich

effects in murine models. BMC Complementary and Alternative Medicine. 2017;**17**(231):1-12

[110] Boakye-Gyasi E, Henneh IT, Abotsi WKM, Ameyaw EO, Woode E. Possible mechanisms involved in the anti-nociceptive effects of hydroethanolic leaf extract of *Ziziphus abyssinica*. Pharmaceutical Biology.

2017;**55**(1):1962-1971

2018;**29**(6):659-669

of an ethanolic extract and xylopic acid from the fruits of *Xylopia aethiopica* in murine models of neuropathic pain. Pharmacognosy Research.

[101] Ameyaw E, Boampong J, Kukuia K, Amoateng P, Obese E, Osei-Sarpong C,

paclitaxel-induced neuropathic pain in rats. Journal of Medical and Biomedical

et al. Effect of xylopic acid on

[102] Ofori AE, Eric W, Samuel K, Robert BP, Nyarko BJ. Anti-nociceptive synergism of pregabalin and xylopic acid co-administration in paclitaxel induced neuropathy: Isobolographic analysis. Pharmacognosy Journal.

Sciences. 2013;**2**(4):6-12

2015;**7**(6):363-368

[103] Woode E, Ameyaw EO,

[104] Woode E, Ameyaw EO, Ainooson G, Abotsi W, Gyasi E, Kyekyeku J. Analgesic effects of an ethanol extract of the fruits of *Xylopia aethiopica* and xylopic acid in murine models of pain: Possible mechanism(s). Pharmacologia. 2013;**4**(4):285-300

[105] Obiri DD, Osafo N. Aqueous ethanol extract of the fruit of *Xylopia aethiopica* (Annonaceae) exhibits anti-anaphylactic and anti-inflammatory actions in mice. Journal of Ethnopharmacology.

2013;**148**(3):940-945

2014;**152**(3):522-531

[106] Obiri DD, Osafo N, Ayande PG, Antwi AO. *Xylopia aethiopica* (Annonaceae) fruit extract suppresses Freund's adjuvant-induced arthritis in Sprague-Dawley rats. Journal of Ethnopharmacology.

Boakye-Gyasi E, Abotsi WKM, Oppong Kyekyeku J, Adosraku R, et al. Effects of an ethanol extract and the diterpene, xylopic acid, of *Xylopia aethiopica* fruits in murine models of musculoskeletal pain. Pharmaceutical Biology. 2016;**54**(12):2978-2986

2014;**6**(2):172-179

**24**

## Antimicrobial Potential of Genes from Garlic (*Allium sativum* L.)

*Hafiz Muhammad Khalid Abbas, Xi Kong, Jia Wu, Mohsin Ali and Wubei Dong*

### **Abstract**

With the advancements in agriculture, farming community less or more started to rely on synthetic chemicals to increase the crop production and protection. But the negative impact of these chemicals on environment and cropping system urges the scientists to discover some new ways to combat with crop disease. By keeping in view, garlic is a well-known economically important vegetable throughout the world and recognized as reservoir for a number of bioactive compounds to treat various diseases; scientists have developed a strategy to identify and isolate antimicrobial genes from garlic. By using *B. subtilis* expression systems, a total of 48 antimicrobial genes, including *AsR 416*, were identified with the potential to significantly retard the growth of economically important fungal and bacterial pathogens. Furthermore, these antimicrobial genes exhibited the thermal stability along with nontoxic effects on mammalian blood cells, which indicate its potential use in the development of human medicines. These genes can revolutionize the way to treat with pathogens and also give a new wave of knowledge to explore the other organisms for the search of antimicrobial genes. This will also help to search the other cost-effective ways for the treatment of plant and human diseases.

**Keywords:** *Allium sativum* L., antimicrobial peptide, *Bacillus subtilis*, resistance gene

### **1. Introduction**

Garlic (*Allium sativum* L.) is one of the most important species of the genus *Allium* and recognized as economically important vegetable throughout the world, especially around the Mediterranean basin where it is considered as main agricultural product [1, 2]. It is also of great importance because of its therapeutic properties and health-related benefits against various kinds of diseases such as aches, deafness, diarrhea, constipation, tumors, and respiratory problems. Health benefits from *Allium* species, especially garlic, have been used for centuries to treat various kinds of disorders, and still, there is need of research to explore its healthrelated potential [3–5]. It is a historic medicinal plant, originated from central Asia about 6000 years ago, and had been started to use as medicine in India since 5000 years ago and 3000 years ago in China [6, 7]. Volatile sulfur compounds, especially thiosulfates, responsible for pungent aroma, are the main compounds

responsible for its physiological effects [8]. Because of its health benefits, garlic is usually recommended as dietary supplement.

During the past few decades, antimicrobial resistance has become one of the most serious and challenging threat for the prevention and treatment of the infectious diseases [9, 10]. Nowadays, much of the attention has been paid to search some new and natural therapeutic agents, which can be used to treat human diseases with high efficacy and minimum adverse effects [11, 12]. Recent advances in research have revealed that there are several natural products with the potential to eliminate or alleviate several serious human diseases, especially cardiovascular, neurodegeneration, cancer, and several other important diseases [13–15]. A large number of researches have elaborated several herbs with the ability to produce antimicrobial compounds as their defense response against the number of different stresses including microbes [16, 17].

With the advancements in agriculture, farming community started to rely more on synthetic chemicals, which have been considered as an important source for crop production and protection. But, hazardous effects of these synthetic chemicals to environment and cropping system make their use questionable [18, 19]. Besides, pathogens also tended to increase their resistance against these synthetic chemicals and threaten the agriculture sustainability [20, 21]. By keeping these challenges in view, the need of identifying new strategies as an alternative source is increasing interestingly. Recently, scientists are trying to understand the chemistry of secondary metabolites from plants, as studies have revealed these secondary metabolites important in several ways, especially allelopathy, biological control, and biofertilizers, and also some compounds have been identified as biostimulants [22–24]. Consequently, understanding the mechanism of these secondary metabolites/ bioactive compounds from plants can be useful for agricultural community.

### **2. Antimicrobial potential of** *Allium* **species**

A number of *Allium* species have antimicrobial potential against variety of microbes including fungi, bacteria, viruses, and other parasites. Among all the *Allium* species, garlic is considered most for antimicrobial research after onion [25].

#### **2.1 Antibacterial potential**

*Allium* extracts containing thiosulfinates have the potential to retard the growth of Gram-positive and Gram-negative bacteria. It is, however, reported that garlic can inhibit the Gram-negative bacteria more than Gram-positive bacteria [26]. The permeability of inhibitory compounds from *Allium* might be affected by the cell wall and cell membrane structure. However, the results were quite opposite with diallyl trisulfide and dimethyl trisulfide and with garlic extracts to conclude that the Gram-positive bacteria were more sensitive than Gram-negative bacteria [27, 28].

Extracts from the garlic are reported to exhibit the effective results against saprobic and pathogenic bacteria, which are resistant to various drugs [29]. Garlic along with ciprofloxacin exhibits the pronounced inhibition of *E. coli* Z17, O2:K1:H- and *Helicobacter pylori*, but no significant evidence was found in the case of *H. pylori* infection in human [30, 31]. It has previously been proved that allicin is the main compound in garlic responsible for the antimicrobial activity, as garlic oil and extracts deficient in allicin do not exhibit any kind of antimicrobial activity [32]. It was later found that garlic oil and its constituting sulfides exhibit the more and significant inhibition of microbes and work as strong antifungal than the antibacterial agent [28].

**27**

garlic.

*Antimicrobial Potential of Genes from Garlic (*Allium sativum *L.)*

up to sulfur number three or four in sulfide molecules [28, 37].

against potato virus Y under in vivo and in vitro conditions [45].

Studies have reported that oils and sulfides from elephant (*A. ampeloprasum*) and shallot (*A. ascalonicum* L.) garlic have the potential to inhibit the food-borne pathogenic bacteria [33, 34]. Ajoene, an unsaturated disulfide, has been reported for its broad-spectrum antibacterial activities, which can be reduced by cysteine, a sulfhydryl compound [35]. Later, it was proved that disulfide in ajoene is a necessary component for the inhibition of bacteria as reduction by sulfhydryl compounds reduces the antibacterial activity. Gram-positive bacteria and yeast are more sensi-

It is reported in different studies that oils and sulfides from the *Allium* have the more potential to inhibit the fungi than bacteria [28, 36]. Antifungal activity of sulfide molecules is directly proportional to increase in the number of sulfur atoms

Another study has also reported that sterilized/autoclaved garlic and its active compounds exhibit significant antifungal activities than that of antibacterial. Further analysis of garlic antimicrobial products revealed that these products are the heterocyclic sulfides [38], allyl alcohol [39], and 3-(allyltrisulfanyl)-2-aminopropanoic acid [40]. For bacteria and yeasts, minimum inhibitory concentrations (MICs) of heterocyclic sulfides are more than 100 and 1–6 ppm [38], respectively, while for the allyl alcohol, 4% and 55–140 ppm MICs are recorded for bacteria and yeasts [39], respectively. In the case of 3-(allyltrisulfanyl)-2-aminopropanoic acid, 100 ppm and less than 50 ppm MICs are observed for bacteria and yeasts [40], respectively. In previous studies, it was mistakenly stated that autoclaved garlic exhibit less antimicrobial activities than fresh garlic. For this statement, the only reason was that they tested autoclaved garlic against bacteria, which was already very less sensitive than yeasts against garlic [41]. Recent studies have explored the germicidal potential of sterilized/

Diallyl polysulfides, as transformation product of allicin, and ajoene exhibit the antiviral activities. From all the reported *Allium* products, it is observed that ajoene exhibits more inhibition than other compounds like allicin and thiosulfinates, but on the other hand, allicin is considered as strong antimicrobial agent [42, 43]. It is thought that antimicrobial compounds from garlic react with viral envelope and inhibit the penetration and exponentiation of influenza virus in animal kidney cells [44]. Garlic aqueous extracts have also been studied to observe the inhibition

A number of parasites, including *Leishmania donovani* [46], *Spironucleus vortens* [47], and *Eimeria papillata* [48], are sensitive to garlic extracts. The MIC values of allicin, dithiins, and ajoene for the inhibition of *S. vortens* growth are higher than the MICs reported for the inhibition of bacteria and fungi, indicating the high toler-

From the above discussed literature, it is clear that garlic has a certain pool of antimicrobial genes which can be isolated and studied further to explore their mechanisms. It will provide some new directions for antimicrobial research. Now we will discuss some techniques to isolate and study the antimicrobial genes from

*DOI: http://dx.doi.org/10.5772/intechopen.83678*

tive to ajoene than Gram-negative bacteria.

**2.2 Antifungal potential**

autoclaved garlic.

**2.3 Antiviral activity**

**2.4 Antiparasitic potential**

ance of *S. vortens* for *Allium* extracts [46].

*Antimicrobial Potential of Genes from Garlic (*Allium sativum *L.) DOI: http://dx.doi.org/10.5772/intechopen.83678*

Studies have reported that oils and sulfides from elephant (*A. ampeloprasum*) and shallot (*A. ascalonicum* L.) garlic have the potential to inhibit the food-borne pathogenic bacteria [33, 34]. Ajoene, an unsaturated disulfide, has been reported for its broad-spectrum antibacterial activities, which can be reduced by cysteine, a sulfhydryl compound [35]. Later, it was proved that disulfide in ajoene is a necessary component for the inhibition of bacteria as reduction by sulfhydryl compounds reduces the antibacterial activity. Gram-positive bacteria and yeast are more sensitive to ajoene than Gram-negative bacteria.

#### **2.2 Antifungal potential**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

usually recommended as dietary supplement.

stresses including microbes [16, 17].

**2. Antimicrobial potential of** *Allium* **species**

**2.1 Antibacterial potential**

antibacterial agent [28].

responsible for its physiological effects [8]. Because of its health benefits, garlic is

During the past few decades, antimicrobial resistance has become one of the most serious and challenging threat for the prevention and treatment of the infectious diseases [9, 10]. Nowadays, much of the attention has been paid to search some new and natural therapeutic agents, which can be used to treat human diseases with high efficacy and minimum adverse effects [11, 12]. Recent advances in research have revealed that there are several natural products with the potential to eliminate or alleviate several serious human diseases, especially cardiovascular, neurodegeneration, cancer, and several other important diseases [13–15]. A large number of researches have elaborated several herbs with the ability to produce antimicrobial compounds as their defense response against the number of different

With the advancements in agriculture, farming community started to rely more on synthetic chemicals, which have been considered as an important source for crop production and protection. But, hazardous effects of these synthetic chemicals to environment and cropping system make their use questionable [18, 19]. Besides, pathogens also tended to increase their resistance against these synthetic chemicals and threaten the agriculture sustainability [20, 21]. By keeping these challenges in view, the need of identifying new strategies as an alternative source is increasing interestingly. Recently, scientists are trying to understand the chemistry of secondary metabolites from plants, as studies have revealed these secondary metabolites important in several ways, especially allelopathy, biological control, and biofertilizers, and also some compounds have been identified as biostimulants [22–24]. Consequently, understanding the mechanism of these secondary metabolites/ bioactive compounds from plants can be useful for agricultural community.

A number of *Allium* species have antimicrobial potential against variety of microbes including fungi, bacteria, viruses, and other parasites. Among all the *Allium* species, garlic is considered most for antimicrobial research after onion [25].

*Allium* extracts containing thiosulfinates have the potential to retard the growth of Gram-positive and Gram-negative bacteria. It is, however, reported that garlic can inhibit the Gram-negative bacteria more than Gram-positive bacteria [26]. The permeability of inhibitory compounds from *Allium* might be affected by the cell wall and cell membrane structure. However, the results were quite opposite with diallyl trisulfide and dimethyl trisulfide and with garlic extracts to conclude that the Gram-positive bacteria were more sensitive than Gram-negative bacteria [27, 28]. Extracts from the garlic are reported to exhibit the effective results against saprobic and pathogenic bacteria, which are resistant to various drugs [29]. Garlic along with ciprofloxacin exhibits the pronounced inhibition of *E. coli* Z17, O2:K1:H- and *Helicobacter pylori*, but no significant evidence was found in the case of *H. pylori* infection in human [30, 31]. It has previously been proved that allicin is the main compound in garlic responsible for the antimicrobial activity, as garlic oil and extracts deficient in allicin do not exhibit any kind of antimicrobial activity [32]. It was later found that garlic oil and its constituting sulfides exhibit the more and significant inhibition of microbes and work as strong antifungal than the

**26**

It is reported in different studies that oils and sulfides from the *Allium* have the more potential to inhibit the fungi than bacteria [28, 36]. Antifungal activity of sulfide molecules is directly proportional to increase in the number of sulfur atoms up to sulfur number three or four in sulfide molecules [28, 37].

Another study has also reported that sterilized/autoclaved garlic and its active compounds exhibit significant antifungal activities than that of antibacterial. Further analysis of garlic antimicrobial products revealed that these products are the heterocyclic sulfides [38], allyl alcohol [39], and 3-(allyltrisulfanyl)-2-aminopropanoic acid [40]. For bacteria and yeasts, minimum inhibitory concentrations (MICs) of heterocyclic sulfides are more than 100 and 1–6 ppm [38], respectively, while for the allyl alcohol, 4% and 55–140 ppm MICs are recorded for bacteria and yeasts [39], respectively. In the case of 3-(allyltrisulfanyl)-2-aminopropanoic acid, 100 ppm and less than 50 ppm MICs are observed for bacteria and yeasts [40], respectively. In previous studies, it was mistakenly stated that autoclaved garlic exhibit less antimicrobial activities than fresh garlic. For this statement, the only reason was that they tested autoclaved garlic against bacteria, which was already very less sensitive than yeasts against garlic [41]. Recent studies have explored the germicidal potential of sterilized/ autoclaved garlic.

#### **2.3 Antiviral activity**

Diallyl polysulfides, as transformation product of allicin, and ajoene exhibit the antiviral activities. From all the reported *Allium* products, it is observed that ajoene exhibits more inhibition than other compounds like allicin and thiosulfinates, but on the other hand, allicin is considered as strong antimicrobial agent [42, 43]. It is thought that antimicrobial compounds from garlic react with viral envelope and inhibit the penetration and exponentiation of influenza virus in animal kidney cells [44]. Garlic aqueous extracts have also been studied to observe the inhibition against potato virus Y under in vivo and in vitro conditions [45].

#### **2.4 Antiparasitic potential**

A number of parasites, including *Leishmania donovani* [46], *Spironucleus vortens* [47], and *Eimeria papillata* [48], are sensitive to garlic extracts. The MIC values of allicin, dithiins, and ajoene for the inhibition of *S. vortens* growth are higher than the MICs reported for the inhibition of bacteria and fungi, indicating the high tolerance of *S. vortens* for *Allium* extracts [46].

From the above discussed literature, it is clear that garlic has a certain pool of antimicrobial genes which can be isolated and studied further to explore their mechanisms. It will provide some new directions for antimicrobial research. Now we will discuss some techniques to isolate and study the antimicrobial genes from garlic.

## **3. Systems for the isolation of antimicrobial genes from garlic**

### **3.1** *Bacillus subtilis* **and** *Escherichia coli* **expression systems**

An experiment was designed to study the antimicrobial genes from the garlic. For this purpose, cDNA libraries from garlic were constructed by using two different vectors, pBE-s and pET22 (b), and then transformed into expression systems, *B. subtilis* and *E. coli*, respectively. For the library quality analysis, two parameters were considered, recombination rate and library titer [49]. For the *E. coli* expression system, 96.7% and 4.6 × 106 pfu/ml, recombination rate and library titer were observed, respectively. On the other hand, recombination rate and library titer for *B. subtilis* expression system were 91.7% and 7.8 × 106 pfu/ml, respectively. Quality analysis revealed gene library in *E. coli* expression system was marginally better than that of the *B. subtilis* expression system.

For the screening of libraries, it was considered that because of the toxicity of protein products of cDNA libraries, *B. subtilis* and *E. coli* cells would be showing autolysis to indicate the antimicrobial potential of these libraries' inserts. For more confirmation, trypan blue dye was also used to indicate the viability of *E. coli* cells [50]. By using this strategy, a number of antimicrobial genes were screened from garlic to reveal its further potentials. For example, in case of *B. subtilis* expression system, a total of 48 antimicrobial genes were screened, including *AsR 416*, while *AsRE 67* was identified by using *E. coli* expression system [51].

### **3.2 Antimicrobial potential of genes from** *A. sativum*

Antimicrobial potential of *A. sativum* genes was studied against fungi and Gram-positive and Gram-negative bacteria [50], and the results were observed as follows (**Tables 1–3**).

### **3.3 Action mechanism of antimicrobial proteins**

A study was designed to explore the action mechanism of antimicrobial peptides. In this study, *B. subtilis* cells were treated with antimicrobial peptide, AsR 416,


**29**

**Table 3.**

*Antimicrobial Potential of Genes from Garlic (*Allium sativum *L.)*

*oryzicola*

**Gram-negative bacteria**

*Agrobacterium tumefaciens*

**Gram-positive bacteria**

*WB800* **—** + **—** + + **— —** *AsR 379* + + + + + + + *AsR 117* + + + + + + + *AsR 412* + + + + + + + *AsR 416* + + + + + + + *AsR 453* + + + + + + + *AsR 36* + + + + + + **—** *AsR 174* + + + + + + **—** *AsR 864* + + + + + + + *AsR 498* + + + + + + + *AsR 845* + + + + + + **—** *AsR 853* + + + + + + +

*C. fangii B. anthracis B. subtilis* **330–2** *B. cereus B. subtilis* **168** *B. subtilis*

*WB800* **— — —** + *AsR 379* **— — —** + *AsR 117* **— — —** + *AsR 412* **— — —** + *AsR 416* **— — —** + *AsR 453* **— — —** + *AsR 36* **— — —** + *AsR 174* **— — —** + *AsR 864* **— — —** + *AsR 498* **— — —** + *AsR 845* **— — —** + *AsR 853* **— — —** +

*E. coli DE3*

*Ralstonia solanacearum*

**WB800**

and then PI (propidium iodide) staining was performed [51]. PI is fluorescent agent that has the ability to bind with DNA through broken cell membrane. Red fluorescence in all bacterial cells treated with antimicrobial peptide was observed under confocal laser microscope [52], while the flow cytometry analysis revealed that cell membrane damages increase with increase in the protein concentration [53].

*DOI: http://dx.doi.org/10.5772/intechopen.83678*

**Genes** *Xanthomonas campestris* **pv***.* 

**—***, indicate no inhibition, +, indicate inhibition.*

*Antimicrobial potential of A. sativum genes against Gram-negative bacteria.*

**Table 2.**

**Genes** *Clavibacter* 

*michiganensis* **subsp.**

**—***, indicate no inhibition, +, indicate inhibition.*

*Antimicrobial potential of A. sativum genes against Gram-positive bacteria.*

#### **Table 1.**

*Antimicrobial potential of A. sativum genes against fungi.*


#### *Antimicrobial Potential of Genes from Garlic (*Allium sativum *L.) DOI: http://dx.doi.org/10.5772/intechopen.83678*

#### **Table 2.**

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

*B. subtilis* expression system were 91.7% and 7.8 × 106

*AsRE 67* was identified by using *E. coli* expression system [51].

**3.2 Antimicrobial potential of genes from** *A. sativum*

**3.3 Action mechanism of antimicrobial proteins**

**—***, indicate no inhibition, +, indicate inhibition.*

*Antimicrobial potential of A. sativum genes against fungi.*

than that of the *B. subtilis* expression system.

sion system, 96.7% and 4.6 × 106

follows (**Tables 1–3**).

**3. Systems for the isolation of antimicrobial genes from garlic**

An experiment was designed to study the antimicrobial genes from the garlic. For this purpose, cDNA libraries from garlic were constructed by using two different vectors, pBE-s and pET22 (b), and then transformed into expression systems, *B. subtilis* and *E. coli*, respectively. For the library quality analysis, two parameters were considered, recombination rate and library titer [49]. For the *E. coli* expres-

observed, respectively. On the other hand, recombination rate and library titer for

For the screening of libraries, it was considered that because of the toxicity of protein products of cDNA libraries, *B. subtilis* and *E. coli* cells would be showing autolysis to indicate the antimicrobial potential of these libraries' inserts. For more confirmation, trypan blue dye was also used to indicate the viability of *E. coli* cells [50]. By using this strategy, a number of antimicrobial genes were screened from garlic to reveal its further potentials. For example, in case of *B. subtilis* expression system, a total of 48 antimicrobial genes were screened, including *AsR 416*, while

analysis revealed gene library in *E. coli* expression system was marginally better

Antimicrobial potential of *A. sativum* genes was studied against fungi and Gram-positive and Gram-negative bacteria [50], and the results were observed as

A study was designed to explore the action mechanism of antimicrobial peptides. In this study, *B. subtilis* cells were treated with antimicrobial peptide, AsR 416,

**Genes** *Fusarium* **spp.** *Botrytis cinerea Phytophthora capsici*

*WB800* **— —** + *AsR 379* **— — —** *AsR 117* **— —** + *AsR 412* **— — —** *AsR 416* **— — —** *AsR 453* **— —** + *AsR 36* **— — —** *AsR 174* **— — —** *AsR 864* **— — —** *AsR 498* **— — —** *AsR 845* **— —** + *AsR 853* **— — —**

pfu/ml, recombination rate and library titer were

pfu/ml, respectively. Quality

**3.1** *Bacillus subtilis* **and** *Escherichia coli* **expression systems**

**28**

**Table 1.**

*Antimicrobial potential of A. sativum genes against Gram-negative bacteria.*


#### **Table 3.**

*Antimicrobial potential of A. sativum genes against Gram-positive bacteria.*

and then PI (propidium iodide) staining was performed [51]. PI is fluorescent agent that has the ability to bind with DNA through broken cell membrane. Red fluorescence in all bacterial cells treated with antimicrobial peptide was observed under confocal laser microscope [52], while the flow cytometry analysis revealed that cell membrane damages increase with increase in the protein concentration [53].

All findings collectively support that the target of antimicrobial peptide is to destroy the cell membrane of target bacteria.

### **3.4 Thermal stability and safety analysis of antimicrobial proteins**

Proteins from *AsR 117*, *AsR 416*, and *AsR 498* were heated at different temperatures for 15 min, and it was found that *AsR 117* and *AsR 416* proteins were thermally stable at all temperature ranges, while *AsR 498* became thermally unstable after 50°C, as it exhibited the reduced antimicrobial activity. For the safety analysis, these proteins were analyzed against sheep red blood cells [54, 55]. This analysis revealed these antimicrobial proteins as nontoxic to mammalian cells with maximum 1000 μg/ml concentration [51]. From the thermal and safety analyses, it is also obvious that antimicrobial genes from garlic can also be used in human medicines in the future, which needs further investigations.

### **4. Conclusion**

It is an adverse need of modern agriculture to search cost-effective ways to treat the crop diseases, as the potential use of synthetic chemicals also increases the resistance in pathogens. Garlic is a famous vegetable for its potential to treat various kinds of diseases. So, it is obvious that antimicrobial genes from garlic are the best source to incorporate resistance in plants without affecting the other environmental factors. This way of introducing resistance can also help to understand the mechanisms of plant biology to further explore the new strategies.

## **Acknowledgements**

This work was supported by the National Major Project for Transgenic Organism Breeding (2011ZX08003-001 and 2016ZX08003-001) and the Hubei Provincial Technology Innovation Program (2016ABA093).

**31**

**Author details**

provided the original work is properly cited.

*Antimicrobial Potential of Genes from Garlic (*Allium sativum *L.)*

*DOI: http://dx.doi.org/10.5772/intechopen.83678*

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

Hafiz Muhammad Khalid Abbas, Xi Kong, Jia Wu, Mohsin Ali and Wubei Dong\* Department of Plant Pathology, College of Plant Science and Technology and the Key Lab of Crop Disease Monitoring & Safety Control in Hubei Province,

Huazhong Agricultural University, Wuhan, Hubei Province, China

\*Address all correspondence to: dwb@mail.hzau.edu.cn

## **Conflict of interest**

The authors declare that they have no conflict of interest.

*Antimicrobial Potential of Genes from Garlic (*Allium sativum *L.) DOI: http://dx.doi.org/10.5772/intechopen.83678*

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

destroy the cell membrane of target bacteria.

the future, which needs further investigations.

**4. Conclusion**

**Acknowledgements**

**Conflict of interest**

All findings collectively support that the target of antimicrobial peptide is to

Proteins from *AsR 117*, *AsR 416*, and *AsR 498* were heated at different temperatures for 15 min, and it was found that *AsR 117* and *AsR 416* proteins were thermally stable at all temperature ranges, while *AsR 498* became thermally unstable after 50°C, as it exhibited the reduced antimicrobial activity. For the safety analysis, these proteins were analyzed against sheep red blood cells [54, 55]. This analysis revealed these antimicrobial proteins as nontoxic to mammalian cells with maximum 1000 μg/ml concentration [51]. From the thermal and safety analyses, it is also obvious that antimicrobial genes from garlic can also be used in human medicines in

It is an adverse need of modern agriculture to search cost-effective ways to treat

This work was supported by the National Major Project for Transgenic Organism

Breeding (2011ZX08003-001 and 2016ZX08003-001) and the Hubei Provincial

the crop diseases, as the potential use of synthetic chemicals also increases the resistance in pathogens. Garlic is a famous vegetable for its potential to treat various kinds of diseases. So, it is obvious that antimicrobial genes from garlic are the best source to incorporate resistance in plants without affecting the other environmental factors. This way of introducing resistance can also help to understand the mecha-

nisms of plant biology to further explore the new strategies.

The authors declare that they have no conflict of interest.

Technology Innovation Program (2016ABA093).

**3.4 Thermal stability and safety analysis of antimicrobial proteins**

**30**

## **Author details**

Hafiz Muhammad Khalid Abbas, Xi Kong, Jia Wu, Mohsin Ali and Wubei Dong\* Department of Plant Pathology, College of Plant Science and Technology and the Key Lab of Crop Disease Monitoring & Safety Control in Hubei Province, Huazhong Agricultural University, Wuhan, Hubei Province, China

\*Address all correspondence to: dwb@mail.hzau.edu.cn

© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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[13] Vasanthi HR, ShriShriMal N, Das DK. Phytochemicals from plants to combat cardiovascular disease. Current Medicinal Chemistry. 2012;**19**(14): 2242-2251. Available from: http:// www.eurekaselect.com/openurl/ content.php?genre=article&issn=0929- 8673&volume=19&issue=14&sp age=2242

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*Antimicrobial Potential of Genes from Garlic (*Allium sativum *L.)*

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[25] Kyung KH. Antimicrobial properties of allium species. Current Opinion in Biotechnology. 2012;**23**(2):142-147. Available from: https://www.sciencedirect.

[26] Perry CC, Weatherly M, Beale T, Randriamahefa A. Atomic force microscopy study of the antimicrobial activity of aqueous garlic versus ampicillin against *Escherichia coli* and *Staphylococcus aureus*. Journal of the Science of Food and Agriculture.

[27] Fujisawa H, Watanabe K, Suma K, Origuchi K, Matsufuji H, Seki T, et al. Antibacterial potential of garlic-derived allicin and its cancellation by sulfhydryl compounds. Bioscience, Biotechnology,

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10.4315/0362-028X-67.3.499

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[19] Hadi F, Hussain F, Hussain M, Ahmad A, Ur Rahman S, Ali N. Phytoextraction of Pb and Cd; the effect of urea and EDTA on *Cannabis sativa* growth under metals stress. International Journal of Agricultural

[20] Talukdar PK, Rahman M, Rahman M, Nabi A, Islam Z, Hoque MM, et al. Antimicrobial resistance, virulence factors and genetic diversity of

*Escherichia coli* isolates from household water supply in Dhaka, Bangladesh.

[21] Kahmann R, Basse C. Fungal gene expression during pathogenesisrelated development and host plant colonization. Current Opinion in Microbiology. 2001;**4**:374-380

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[15] Nabavi SF, Russo GL, Daglia M, Nabavi SM. Role of quercetin as an alternative for obesity treatment: You are what you eat! Food Chemistry. 2015;**179**:305-310

[16] Bednarek P. Chemical warfare or modulators of defence responses—The function of secondary metabolites in plant immunity. Current Opinion in Plant Biology. 2012;**15**:407-414

[17] Daglia M. Polyphenols as antimicrobial agents. Current Opinion in Biotechnology. 2012;**23**:174-181

[18] Sugeng AJ, Beamer PI, Lutz EA, Rosales CB. Hazard-ranking of agricultural pesticides for chronic health effects in Yuma County, Arizona. Science of the Total Environment. 2013;**463-464**:35-41

[19] Hadi F, Hussain F, Hussain M, Ahmad A, Ur Rahman S, Ali N. Phytoextraction of Pb and Cd; the effect of urea and EDTA on *Cannabis sativa* growth under metals stress. International Journal of Agricultural Research. 2014

[20] Talukdar PK, Rahman M, Rahman M, Nabi A, Islam Z, Hoque MM, et al. Antimicrobial resistance, virulence factors and genetic diversity of *Escherichia coli* isolates from household water supply in Dhaka, Bangladesh. PLoS One. 2013;**8**(4)

[21] Kahmann R, Basse C. Fungal gene expression during pathogenesisrelated development and host plant colonization. Current Opinion in Microbiology. 2001;**4**:374-380

[22] Gniazdowska A, Bogatek R. Allelopathic interactions between plants. Multi site action of allelochemicals. Acta Physiologiae Plantarum. 2005;**27**:395-407

[23] Cantor A, Hale A, Aaron J, Traw MB, Kalisz S. Low allelochemical concentrations detected in garlic mustard-invaded forest soils inhibit fungal growth and AMF spore germination. Biological Invasions. 2011;**13**(12):3015-3025

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[25] Kyung KH. Antimicrobial properties of allium species. Current Opinion in Biotechnology. 2012;**23**(2):142-147. Available from: https://www.sciencedirect. com/science/article/pii/ S0958166911006720

[26] Perry CC, Weatherly M, Beale T, Randriamahefa A. Atomic force microscopy study of the antimicrobial activity of aqueous garlic versus ampicillin against *Escherichia coli* and *Staphylococcus aureus*. Journal of the Science of Food and Agriculture. 2009;**89**(6):958-964

[27] Fujisawa H, Watanabe K, Suma K, Origuchi K, Matsufuji H, Seki T, et al. Antibacterial potential of garlic-derived allicin and its cancellation by sulfhydryl compounds. Bioscience, Biotechnology, and Biochemistry. 2009;**73**(9): 1948-1955. Available from: http://www. tandfonline.com/doi/full/10.1271/ bbb.90096

[28] Kim JW, Kim YS, Kyung KH. Inhibitory activity of essential oils of garlic and onion against bacteria and yeasts. Journal of Food Protection. 2004;**67**(3):499-504. DOI: 10.4315/0362-028X-67.3.499

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[12] Cooper AI, Poliakoff M. Highpressure reactions in polyethylene films, a new development in matrix isolation. The photochemical reaction of Fe(CO)5 with N2 and the thermal reaction of Fe(CO)4 (N2) with H2. Chemical Physics Letters.

[13] Vasanthi HR, ShriShriMal N, Das DK. Phytochemicals from plants to combat cardiovascular disease. Current Medicinal Chemistry. 2012;**19**(14): 2242-2251. Available from: http:// www.eurekaselect.com/openurl/ content.php?genre=article&issn=0929- 8673&volume=19&issue=14&sp

1993;**212**(6):611-616

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[52] Xie J, Gou Y, Zhao Q , Wang K, Yang X, Yan J, et al. Antimicrobial activities and membrane-active mechanism of CPF-C1 against multidrug-resistant bacteria, a novel antimicrobial peptide derived from skin secretions of the tetraploid frog *Xenopus clivii*. 2014. DOI:

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ume=14&issue=2&spage=233

[44] Mehrbod P, Aini I, Amini E, et al. Assessment of direct immunofluorescence assay in detection of antiviral effect of garlic extract on influenza virus. African Journal of Microbiology Research. 2013;**7**(21):2608-2612. Available from: http://academicjournals.org/journal/ AJMR/article-abstract/C7BAEBF12939

[45] Mohamed EF. Antiviral properties of garlic cloves juice compared with onion bulbs juice against potato virus Y (PVY). Journal of American Science. 2010;**6**. Available from: http:// www.americanscience.orgeditor@

[46] Sharma U, Velpandian T, Sharma

leishmanial activity of selected Indian plants known to have antimicrobial properties. Parasitology Research.

[47] Millet COM, Lloyd D, Williams C, Williams D, Evans G, Saunders RA, et al. Effect of garlic and allium-derived products on the growth and metabolism of *Spironucleus vortens*. Experimental Parasitology. 2011;**127**(2):490-499

[48] Dkhil MA, Abdel-Baki AS, Wunderlich F, Sies H, Al-Quraishy S. Anticoccidial and antiinflammatory activity of garlic in murine *Eimeria papillata* infections. Veterinary Parasitology. 2011;**175**(1-2):66-72

americanscience.org302

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P, Singh S. Evaluation of anti-

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*Medicinal Plants - Use in Prevention and Treatment of Diseases*

[35] Terzioǧlu S, Başkent EZ, Sivrikaya F, Çakir G, Kadioǧullari AI, Başkaya Ş, et al. Monitoring forest plant biodiversity changes and developing conservation strategies: A study from Camili Biosphere Reserve area in NE Turkey. Biologia (Bratislava).

2010;**65**(5):843-852

[36] Mahmoudabadi AZ, Nasery MKG. Antifungal activity of shallot, *Allium ascalonicum* Linn. (Liliaceae), in vitro. Journal of Medicinal Plants Research. 2009;**35**(12):2531-2541

[37] O'Gara EA, Hill DJ, Maslin DJ. Activities of garlic oil, garlic powder,

[38] Choi MK, Chae K-Y, Lee J-Y, Kyung KH. Antimicrobial activity of chemical substances derived from S-alk(en) yl-L-cysteine sulfoxide (alllin) in garlic, *Allium sativum* L. Food Science and Biotechnology. 2007;**16**(1):1-7

[39] Kyung KH, Choi JH. Allyl alcohol is the sole antiyeast compound in heated garlic extract. Journal of Food Science.

[40] Kang S-S, Lim DR, Kyung KH. 3-(Allyltrisulfanyl)-2-aminopropanoic acid, a novel nonvolatile water-soluble antimicrobial sulfur compound in heated garlic. Journal of Medicinal Food. 2010;**13**. DOI: 10.1089/

[41] Walton L, Herbold M, Lindegren CC. Bactericidal effects of vapors from crushed garlic. Journal of Food Science. 1936;1(2):163-169. Available from: 10.1111/j.1365-2621.1936.tb17778.x

[42] Weber ND, Andersen DO, North JA, Murray BK, Lawson LD, Hughes BG. In vitro virucidal effects of *Allium sativum* (garlic) extract and compounds. Planta Medica. 1992;**58**(5):417-423. Available

and their diallyl constituents against *Helicobacter pylori*. Applied and Environmental Microbiology.

2000;**66**(5):2269-2273

2005;**70**(6):305-309

jmf.2010.1059

[29] Dubey D, Rath S, Sahu MC, Debata NK, Padhy RN. Antimicrobials

of plant origin against TB and other infections and economics of plant drugs—Introspection. Indian Journal of Traditional Knowledge.

[30] Graham DY, Anderson SY, Lang T. Garlic or jalapeno peppers for treatment of *Helicobacter pylori* infection. The American Journal of Gastroenterology. 1999;**94**(5):1200-1202

article/pii/S0924857909000971

Medicine. 2014;**14**(1)

[33] Rattanachaikunsopon P, Phumkhachorn P. Antimicrobial activity of elephant garlic oil against *Vibrio cholerae* in vitro and in a food model. Bioscience, Biotechnology, and Biochemistry. 2009;**73**(7):1623-1627.

DOI: 10.1271/bbb.90128

[34] Rattanachaikunsopon P, Phumkhachorn P. Shallot (*Allium* 

and antimicrobial activity against food-borne pathogenic bacteria. African Journal of Microbiological Research. 2009;**3**(11):747-750. Available from: http://www.academicjournals.org/ajmr

*ascalonicum* L.) oil: Diallyl sulfide content

[32] Booyens J, Thantsha MS. Fourier transform infra-red spectroscopy and flow cytometric assessment of the antibacterial mechanism of action of aqueous extract of garlic (*Allium sativum*) against selected probiotic Bifidobacterium strains. BMC Complementary and Alternative

[31] Sohn DW, Han CH, Jung YS, Kim SI, Kim SW, Cho YH. Anti-inflammatory and antimicrobial effects of garlic and synergistic effect between garlic and ciprofloxacin in a chronic bacterial prostatitis rat model. International Journal of Antimicrobial Agents. 2009;**34**(3):215-219. Available from: https://www.sciencedirect.com/science/

2012;**11**(2):225-233

**34**

[43] Schäfer G, Kaschula C. The immunomodulation and antiinflammatory effects of garlic organosulfur compounds in cancer chemoprevention. Anti-Cancer Agents in Medicinal Chemistry. 2014;**14**(2):233- 240. Available from: http://www. eurekaselect.com/openurl/content. php?genre=article&issn=1871-5206&vol ume=14&issue=2&spage=233

[44] Mehrbod P, Aini I, Amini E, et al. Assessment of direct immunofluorescence assay in detection of antiviral effect of garlic extract on influenza virus. African Journal of Microbiology Research. 2013;**7**(21):2608-2612. Available from: http://academicjournals.org/journal/ AJMR/article-abstract/C7BAEBF12939

[45] Mohamed EF. Antiviral properties of garlic cloves juice compared with onion bulbs juice against potato virus Y (PVY). Journal of American Science. 2010;**6**. Available from: http:// www.americanscience.orgeditor@ americanscience.org302

[46] Sharma U, Velpandian T, Sharma P, Singh S. Evaluation of antileishmanial activity of selected Indian plants known to have antimicrobial properties. Parasitology Research. 2009;**105**(5):1287-1293

[47] Millet COM, Lloyd D, Williams C, Williams D, Evans G, Saunders RA, et al. Effect of garlic and allium-derived products on the growth and metabolism of *Spironucleus vortens*. Experimental Parasitology. 2011;**127**(2):490-499

[48] Dkhil MA, Abdel-Baki AS, Wunderlich F, Sies H, Al-Quraishy S. Anticoccidial and antiinflammatory activity of garlic in murine *Eimeria papillata* infections. Veterinary Parasitology. 2011;**175**(1-2):66-72

[49] Park NJ, Zhou X, Yu T, Brinkman BMN, Zimmermann BG, Palanisamy V, et al. Characterization of salivary RNA by cDNA library analysis. Archives of Oral Biology. 2007;**52**(1):30-35. Available from: http://www.ncbi.nlm. nih.gov/pubmed/17052683

[50] Cheng X, Liu G, Ye G, Wang H, Shen X, Wu K, et al. Screening and cloning of antimicrobial DNA sequences using a vital staining method. Gene. 2009;**430**((1-2)): 132-139. Available from: https://www. sciencedirect.com/science/article/pii/ S0378111908005453

[51] Kong X, Yang M, Abbas HMK, Wu J, Li M, Dong W. Antimicrobial genes from *Allium sativum* and *Pinellia ternata* revealed by a *Bacillus subtilis* expression system. Scientific Reports. 2018;**8**(1):14514. Available from: http://www.nature.com/articles/ s41598-018-32852-x

[52] Xie J, Gou Y, Zhao Q , Wang K, Yang X, Yan J, et al. Antimicrobial activities and membrane-active mechanism of CPF-C1 against multidrug-resistant bacteria, a novel antimicrobial peptide derived from skin secretions of the tetraploid frog *Xenopus clivii*. 2014. DOI: 10.1002/psc.2679

[53] Lee H, Hwang J-S, Lee J, Kim JI, Lee DG. Scolopendin 2, a cationic antimicrobial peptide from centipede, and its membrane-active mechanism. Biochimica et Biophysica Acta – Biomembranes. 2015;**1848**(2): 634-642. Available from: https://www. sciencedirect.com/science/article/pii/ S0005273614004143

[54] Joshi S, Bisht GS, Rawat DS, Kumar A, Kumar R, Maiti S, et al. Interaction studies of novel cell selective antimicrobial peptides with model membranes and *E. coli* ATCC 11775. Biochimica et Biophysica Acta— Biomembranes. 2010;**1798**(10):

Chapter 3

Abstract

Himalayas

Medicinal Plants Used for

Treatment of Prevalent Diseases

in Northern Pakistan of Western

In this research study, we have scientifically assessed medicinal species and herbal preparations used by inhabitants of Northern Pakistan to treat joint pain, hypertension, skin diseases and glottis infections. The aim of the study is to document and highlight the ethnopharmacological significance and compare the uses of medicinal herbs for curing prevalent ailments in Northern Pakistan. Ethnomedicinal data were collected from 180 informants using semi-structured interviews and group meetings. A total of 80 plant species in 54 families were reported for the treatment of various health conditions. Heliotropium lasiocarpum, Geranium wallichianum, Parkinsonia aculeata, Rubia cordifolia and Salvadora persica were the favored plants for curing these diseases. Highest RFC was recorded for Neolitsea chinensis (0.956), Rubia cordifolia (0.928). The similarity of the informer's knowledge about used medicines was found in Aesculus indica and Abies pindrow with high UV. Cuscuta reflexa and Lawsonia inermis had 98–99% fidelity level for management of joint pain, skin diseases, glottis infection and hypertension respectively. In Northern Pakistan, a rich diversity of medicinal plants was used in curing various diseases. The results of this study help us in screening of herbal plants for further phytochemical and pharmacological study which leads to discovery of natural drug and development with global interest for cure of various ailments.

Keywords: herbaceous diversity, ethnomedicinal, diseases, Northern Pakistan,

Ethnomedicinal literature put emphasis on the relation between the indigenous communities and the usage of plants [1]. Plants are important for all biomes and the working of all social societies [2]. Traditional herbal drugs have been effective as a remedy for wide variety of diseases [3]. Traditional medicinal species and plant derivative treatments are extensively utilized in old medicinal systems worldwide,

herbal preparation, frequency of citation

1.1 Ethnobotany: concept and significance

1. Introduction

37

Khafsa Malik, Mushtaq Ahmad, Muhammad Zafar,

Shazia Sultana, Athar Tariq and Neelam Rashid

1864-1875. Available from: https://www. sciencedirect.com/science/article/pii/ S0005273610002142

[55] Wingfield P. Protein precipitation using ammonium sulfate. Current Protocols in Protein Science. 2001;**3**(3):Appendix 3F. Available from: http://www.ncbi.nlm.nih.gov/ pubmed/18429073

### Chapter 3

*Medicinal Plants - Use in Prevention and Treatment of Diseases*

1864-1875. Available from: https://www. sciencedirect.com/science/article/pii/

[55] Wingfield P. Protein precipitation using ammonium sulfate. Current Protocols in Protein Science. 2001;**3**(3):Appendix 3F. Available from: http://www.ncbi.nlm.nih.gov/

S0005273610002142

pubmed/18429073

**36**
