**5. Fertility and ROS**

Physiological levels of free radicals are essential in the regulation of sperm maturation, capacitation and hyperactivation, acrosome reaction and sperm-oocyte fusion [15]. ROS can make their way into the seminal plasma where they have a regulatory and a damaging an effect on spermatozoa. Leukocytes and immature spermatozoa are sources of ROS within the seminal plasma [15]. Leukocytes produce ROS as part of their mechanism to clear the body of infections [56], but during an infection, increased production and chemotaxis of leukocytes leads to an increase in ROS and a reduction in the antioxidant called superoxide dismutase [57]. This results in oxidative stress which has negative effects on the function of spermatozoa [15]. The changes in sperm parameters brought about are a reduction in motility and in the fertilisation capacity [58, 59]. The presence of excess cytoplasm in immature spermatozoa leads to activation of the NADPH system, which donates electrons to free radicals [13], allowing them to build up within the seminal plasma. This excess of free radicals leads to an increase in ROS and the disadvantages thereof.

Exogenous sources of ROS allow pathological amounts of ROS to build up within the seminal plasma [15]. Environmental pollutants increase ROS in testes, causing oxidative stress, DNA damage and impaired sperm production [15]. Cigarette smoking allows nicotine to make its way into the seminal plasma, which has been associated with the production of free radicals and a decrease in antioxidant function [15]. Cigarette smoking also increases concentrations of leukocytes in seminal plasma [15]. Excessive intake of alcohol increases ROS while decreas‐ ing antioxidants due to increased ethanol metabolism [60]. Spinal cord injury is associated with higher levels of ROS and higher numbers of leukocytes, both of which are known to contribute to oxidative stress, leading to reduces sperm motility and poor sperm morphology [61, 62]. Dilatation of veins in the plexus surrounding the spermatic cord (varicocele) is associated with increased levels of ROS, with a concomitant decrease in sperm function [63]. Higher quantities of nitric oxide have also been found in men with varicocele, and these men have also been found to have higher markers of oxidative stress [64].

increase urinary output [52], serving to reduce colonisation of the urethra by bacteria, reducing

Buchu may be used in the form of a herbal tincture or a herbal tea. The tincture will deliver adequate quantities of the herb allowing its actions to be distributed throughout the body. The tea will support the herbal action of the tincture and simultaneously increase fluid intake [52].

Buchu may be used alone or in combination with other plants. In combination, the synergistic interactions of the extracts found in buchu and those of other plants will produce a better therapeutic effect in the treatment of a variety of ailments, or, in this instance, in the treatment of UTI's[44]. Antibiotic resistance is rife, and is the cause of a prolonged disease course. A combination of antimicrobial agents helps prevent resistance to antibiotics, increases the spectrum of activity and may even reduce the side effects of therapy [44]. A combination of the plants Agathosma crenulata, Dodonaea viscose and Eucalyptus globules produces a broad spectrum and enhanced antimicrobial activity [44]. Combination of these plants in a 1:1:1 ratio provides additive and synergistic effects [53] where the extracts of each plant act via multiple mechanisms to reduce the number of disease causing microbials. The mixture of bioactive constituents target numerous target sites and work in a synergistic manner [53]. The combi‐ nation of these plants give an additive anti-inflammatory, anti-fungal, analgesic, antibacterial

By preventing and reducing colonisation of the urinary tract that leads to urinary tract infections, other co-morbidities associated with UTI's such as erectile dysfunction, prostatitis

Physiological levels of free radicals are essential in the regulation of sperm maturation, capacitation and hyperactivation, acrosome reaction and sperm-oocyte fusion [15]. ROS can make their way into the seminal plasma where they have a regulatory and a damaging an effect on spermatozoa. Leukocytes and immature spermatozoa are sources of ROS within the seminal plasma [15]. Leukocytes produce ROS as part of their mechanism to clear the body of infections [56], but during an infection, increased production and chemotaxis of leukocytes leads to an increase in ROS and a reduction in the antioxidant called superoxide dismutase [57]. This results in oxidative stress which has negative effects on the function of spermatozoa [15]. The changes in sperm parameters brought about are a reduction in motility and in the fertilisation capacity [58, 59]. The presence of excess cytoplasm in immature spermatozoa leads to activation of the NADPH system, which donates electrons to free radicals [13], allowing them to build up within the seminal plasma. This excess of free radicals leads to an increase

Exogenous sources of ROS allow pathological amounts of ROS to build up within the seminal plasma [15]. Environmental pollutants increase ROS in testes, causing oxidative stress, DNA damage and impaired sperm production [15]. Cigarette smoking allows nicotine to make its

and erectile dysfunction may be simultaneously prevented.

the duration of the UTI.

306 Antioxidant-Antidiabetic Agents and Human Health

and antiviral effect [54, 55].

**5. Fertility and ROS**

in ROS and the disadvantages thereof.

As previously stated, ROS have both physiological and pathological roles. Physiological levels of ROS have been found to play a vital role in normal sperm function [15]. ROS are signalling molecules in many processes which allow fertilisation [15]. ROS are essential in sperm maturation as they have been shown to stabilise sperm chromatin, which is required to maintain genetic integrity [15]. Capacitation, a process which occurs in the female genital tract, is a maturation process which gives sperm the ability to gain hyperactive motility and the potential to carry out an acrosome reaction [15]; both of which are vital for fertilisation to take place. Capacitation is a series of molecular changes that brings about membrane hyperpolarisation, protein phosphorylation and an increase in pH and cAMP [65]. Hyperactivation, a subsection of capacitaion, provide spermatozoa with the ability to move through cervical mucous and to penetrate cumulus cells of the oocyte [15]. The acrosome reaction, which is the release of proteolytic enzymes contained within the acrosomal cap, helps to degrade the extracellular matrix of the zona pellucida [65]. This allows spermatozoa to burrow through the zona pellucida, allowing penetration of this layer to permit fusion with the oocyte. Physiological levels of ROS are also essential for sperm-oocyte fusion [15]. Fluidity of the sperm membrane is required to fusion to take place [66]. This is acquired by spermatozoa through capacitation and acrosome reaction [15], processes which are both assisted by the presence of ROS. Membrane fluidity allows sperm to fuse with the oocyte, allowing successful fertilisation.

Pathological levels of ROS are detrimental to sperm function [15]. An imbalance between ROS and antioxidant defence mechanisms results in pathological processes to be initiated. ROS cause damage to proteins, lipid molecules and DNA [67]. Lipid peroxiation causes loss of fatty acids from sperm plasma membranes, which affects the structure and func‐ tion (i.e. fluidity, transport processed, receptor transduction) of spermatozoa, impairing the functional parameters of sperm and their ability to fertilize [67, 68]. Sperm is usually resistant to DNA damage due to the tight packaging of DNA material and the innate antioxidant defense mechanisms in place [69]. ROS causes DNA damage and leads to infertility. Nitric oxide and hydrogen peroxide have been found to induce DNA damage, which brings about DNA fragmentation and reduced sperm parameters (i.e. motility, and morphology) [70, 73]. The impaired the ability of the Y chromosome to repair DNA strand breaks makes spermatozoa more vulnerable to DNA damage [74]. Damage to cellular components initiates apoptosis. Abnormal spermatozoa are regularly removed via program‐

med cell death, which is assisted by the presence of ROS [75]. ROS activate mitochondri‐ al release of cytochrome c, which ultimately activates caspases which leads to apoptosis [76, 78]. Hypochlorous acid is also known to oxidise cellular components, directly activat‐ ing apoptosis [78]. Also, high temperatures within the male reproductive system have been correlated to higher levels of ROS [13].

**Author/Study Findings/Uses of Buchu Application/Mechanism of action**

Buchu – The Multi-Purpose Ethnomedicinally Important Specie and Its Benefits in the Reproductive System

Members of the Agathosma specie contain flavonoids, i.e. diosmin, hesperidin,rutin, quercitin, mucilage and resins with extensive anti-oxidant properties These compounds act via several mechanisms to reduce free radicals

http://dx.doi.org/10.5772/57233

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Buchu essential oils and extracts active against selected pathogens, namely Staphylococcus aureus, Bacillus cereus, Klebsiella pneumonia and Candida albicans. More active against gram positive than gram negative bacteria Affects the development of biofilms by preventing attachment of bacteria Agathosma species found to contain coumarins, phenolic substances with benzene and α-pyrone rings. A number of these compounds have been found to be active against microbials by stimulating macrophages

Oil distilled from Buchu leaves has therapeutic properties and major uses, including the treatment of kidney and urinary tract infections, as well as haematuria and prostatitis

Diospenol is responsible for the diuretic action of Buchu. Diosphenol acts by irritating the gallbladder, causing the production of urine. Buchu contains flavonoids that induce urine production

The Agathosma species has been many applications, among these are its uses in the treatment of arthritis and rheuamtism through its ability to reduce uric acid. Buchu has been listed as a treatment for stomach ailments and for relief from nausea, diarrhoea and flatulence

A number of Buchu preparations are used to deliver it to the body. It may be soaked in vinegar and used as an external application to treat bruises, contusions, sprains and fractures Used as an antiseptic to clean wounds

Anti-oxidant properties

Antibacterial Antimicrobial Antifungal Antibiotic repellent

haematuria calculi kidney disease infections of the bladder, prostate, urethra

Diuretic properties

Arthritis, Rheumatism Nausea and diarrhoea Flatulence

Bruises, contusions, sprains, fractures Cleans wounds

Moolla *et al*, 2007

Moolla and Viljoen, 2009

Cowan MM, 1999

Moolla *et al*, 2007 Lis-Balchin *et al*, 2001 Shwegler, 2003

Viljoen *et al*, 2007

Moolla and Viljoen, 2009

Watt and Breyer-Brandwijk, 1962

Gentry HS, 1961 Eaton CL Simpson, 1988 Moolla *et al*, 2007

Moolla and Viljoen, 2009 Lis-Balchin M *et al*, 2001 Bajpayee P *et al*, 2012 Grieve M, 1937 Grieve M, 1937

Van Wyk *et al*, 1997

Lis-Balchin *et al*, 2001 Urinary problems

Antioxidants have been explored as possible therapeutics that may lead to a reduction in levels of ROS [15]. Antioxidant therapy has been shown to reduce oxidative stress, resulting in reduced DNA damage and apoptosis. Antioxidants may be used to keep free radicals at levels that are appropriate for physiological function, while reducing the levels that bring about unwanted pathologies and infertility [15].

Buchu is well known for its antioxidant properties. As discussed above, it has been shown to contain factors which assist in the antioxidant activity of the plant. By using this plant in small therapeutic doses, Buchu may be effective in staving off excess build up of free radicals, decreasing the negative effects on the function of spermatozoa that lead to infertility.
