Introduction to Antioxidants and Their Importance in Human Health

### **Chapter 1** Antioxidants Sources

*Marjan Assefi, Kai-Uwe Lewandrowski, Sohila Nankali and Alireza Sharafshah*

### **Abstract**

Natural antioxidants are abundant in food and medicinal plants. These natural antioxidants, particularly polyphenols and carotenoids, have numerous biological effects, including anti-inflammatory, anti-aging, anti-atherosclerosis, and anticancer properties. To examine potential cancer prevention agent sources and advance their utilization in useful food varieties, drugs, and food added substances, it is fundamental for separate cell reinforcements from food and restorative plants really and assess them suitably. This paper goes into great detail about the green extraction methods of natural antioxidants, the evaluation of antioxidant activity at the chemical and cellular levels, and their primary sources, which are food and medicinal plants.

**Keywords:** medicinal plants, cellular, prevention, antioxidants, natural, biological effects, anti cancer

### **1. Introduction**

Reactive oxygen species (ROS) and reactive nitrogen species (RNS) like superoxide, hydroxyl, and nitric oxide can damage DNA and oxidize proteins and lipids in cells in a biological system. Typically, the body's cell reinforcement framework can dispose of these extremists, keeping the harmony among oxidation and against oxidation. However, environmental toxins, cigarette smoking, alcohol, radiation, or other forms of exposure can cause excessive ROS and RNS production [1]. These ROS and RNS can cause a variety of chronic and degenerative diseases because they upset the balance between oxidation and antioxidation. The increased intake of exogenous antioxidants would lessen the damage caused by oxidative stress by acting as free radical scavengers, singlet oxygen quenchers, and reducing agents. An oxidative chain reaction would not start or spread as a result of this [2, 3].

The majority of the exogenous antioxidants come from food and medicinal plants like mushrooms, beverages, flowers, spices, and traditional medicinal herbs. In addition, the industries that process agricultural by-products are potential significant natural antioxidant sources. Polyphenols (phenolic acids, flavonoids, anthocyanins, lignans, and stilbenes), carotenoids (xanthophylls, carotenes), and vitamins (vitamins E and C) are the primary natural antioxidants derived from plant materials. In general, these natural antioxidants, particularly polyphenols and carotenoids, have a wide range of biological effects, including anti-aging, anti-cancer, anti-inflammatory, and antibacterial effects [4].

Food science and nutrition are paying a lot of attention to the effective extraction methods of natural antioxidants, the appropriate evaluation of antioxidant activity, and the fact that their primary sources are food and medicinal plants. Ultrasoundassisted extraction, microwave-assisted extraction, enzyme-assisted extraction, pressurized liquid extraction, supercritical fluid extraction, high hydrostatic pressure extraction, pulsed electric field extraction, and high voltage electrical discharges extraction are just a few green non-conventional methods that have been developed to improve the efficiency with which antioxidant components are extracted from plant materials. Additionally, a variety of evaluation assays, such as the Trolox equivalence antioxidant capacity (TEAC) assay, the ferric ion reducing antioxidant power (FRAP) assay, the oxygen radical absorbance capacity (ORAC) assay, the inhibiting the oxidation of low-density lipoprotein (LDL) assay, the cellular antioxidant activity assay, and others, have been developed to further evaluate the antioxidant capacities of extracts from natural products, particularly those that These tests have been used to rank antioxidant plants and suggest the best foods for antioxidant consumption. The purpose of this review is to provide a summary of the methods used to extract natural antioxidants, methods used to evaluate antioxidant activity, and their primary sources, which are food and medicinal plants [5–7].

The type and concentration of the extraction solvent, the extraction temperature, the extraction time, and the extraction pH are just a few of the extraction factors that have a significant impact on the efficiency of the extraction. Antioxidants have been extracted from food and medicinal plants using a variety of solvents. The chemical nature and polarity of the antioxidant compounds to be extracted determine the solvent selection. The majority of the phenolics, flavanoids, and anthocyanins are antioxidants that dissolve in water. Extraction frequently makes use of polar and medium-polar solvents like water, ethanol, methanol, propanol, acetone, and their aqueous mixtures. Carotenoids are antioxidants that dissolve in lipids. For extraction, common organic solvents like mixtures of hexane with acetone, ethanol, and methanol or ethyl acetate with acetone, ethanol, and methanol have been used [8, 9].

Antioxidants can be extracted from food and medicinal plants using a variety of extraction methods, including conventional and non-conventional methods. The

most common conventional extraction methods are hot water bath, maceration, and Soxhlet extraction. These methods take a long time, use a lot of organic solvents, and have low extraction yields. Additionally, the long heating process in hot water bath and Soxhlet extraction may cause thermolabile compounds to break down [10]. Non-conventional techniques like ultrasound, microwave, pressurized liquid, enzyme hydrolysis, supercritical fluids, high hydrostatic pressure, pulsed electric field, and high voltage electrical discharges have been investigated for the purpose of obtaining antioxidants from plants in a way that is both energy efficient and economically sustainable.

### **2. Main resources of natural antioxidants**

The TEAC assay, which measures antioxidants' capacity to scavenge free radicals, the FRAP assay directly measures antioxidants' reducing capacity, and the total phenols assay by FCR evaluates the phenolic contents of tested samples are the primary sources of natural antioxidants [11–13]. The combination of the TEAC, FRAP, and FCR methods is frequently utilized to evaluate the antioxidant activity in order to carry out in-depth research on various aspects of antioxidants. Numerous food and medicinal plants, such as fruits, vegetables, cereal grains, edible and wild flowers, macro-fungi, medicinal plants, spices, and so on, have been widely estimated to have antioxidant properties [14]. A combination of the results from the FRAP, TEAC, and FCR assays was used to identify the varieties with strong antioxidant properties. In general, these findings demonstrated that diverse categories had a wide range of antioxidant capacities. From 0.11 0.01 to 72.11 2.19 mol Fe(II)/g, 0.84 0.03 to 80.68 2.11 mol Trolox/g, and 11.88 0.11 to 585.52 18.59 mg GAE/100 g, respectively, the FRAP, TEAC, and FCR values of 62 fruits varied. 56 vegetables had FRAP, TEAC, and FCR values ranging from 2.69 to 60.9 mol Fe(II)/g, 6.93 to 33.63 mol Trolox/g, and 4.99 to 23.27 mg GAE/g, respectively. From 5.23 0.23 to 126.19 2.91 mol Fe(II)/g, 0.62 0.14 tso 30.03 1.10 mol trolox/g, and 1.35 0.15 to 9.47 0.48 mg GAE/g, respectively,

the FRAP, TEAC, and FCR values of 24 cereal grains varied. From 0.14 to 1844.85 mol Fe(II)/g, 0.99 to 1544.38 mol Trolox/g, and 0.19 to 101.33 mg GAE/g, respectively, the FRAP, TEAC, and FCR values of 223 medicinal plants varied. Clearly, medicinal plants had significantly higher antioxidant activities and total phenolic content than fruits, vegetables, and cereals among these varieties with strong antioxidant properties [11, 15, 16].

Moreover, the cancer prevention agent exercises of food and restorative plants have additionally been assessed by cell reinforcement action measures in light of various cell types. The 27 vegetables' cellular antioxidant activities ranged from not detected (tomato) to 41.9 6.2 mol of QE/100 g (beet). The 25 fruits' cellular antioxidant activities ranged from 3.15 0.21 mol of QE/100 g (banana) to 292 11 mol of QE/100 g (wild blueberry). In the two examinations, these outcomes showed that CAA values were fundamentally connected with absolute phenolic content. Surarit and others based on HL-60 cells, it was reported that the ethanolic bran extracts of 11 Thai red and purple and two non-pigmented rice varieties performed the following cellular antioxidant activities: non-pigmented rice followed by purple rice in the same order as red rice in terms of phenolic and flavonoid content in these rice extracts [17, 18].

Chemical assays cannot completely capture the sample's in vivo behavior when evaluating its antioxidant capacity. Antioxidants must be evaluated for their efficacy under more biologically relevant conditions. Although more expensive and timeconsuming, animal models and human studies are more suitable for evaluation [19]. The cellular antioxidant activity (CAA) assay has been developed to evaluate antioxidant capacities as intermediate testing methods. The Dichlorofluorecin (DCFH) method is a common CAA assay that measures antioxidants' ability to prevent DCFH oxidation. In human hepatocarcinoma HepG2 cells, ABAP-generated peroxyl radicals easily convert DCFH that is trapped within the cells to fluorescent dichlorofluorescein (DCF). Fluorescence could be used to monitor DCF (exc = 485 nm, em = 538 nm). The antioxidant capacity of bioactive components is inversely proportional to the decrease in cellular fluorescence [20–23]. Human red blood cells, human endothelial EA.hy926, human colon cancer Caco-2 cells, human macrophage U937 cells, and mouse macrophage RAW264.7 cells have all been utilized for the CAA assay, with the exception of HepG2 cells. Additionally, a microfluidic cell chip-based CAA assay with arrayed microchannels has been developed to evaluate plant antioxidants. There are 48 distinct parallel array channels and 288 round cell culture micro chambers on the microfluidic chip. With this method, a multimode reader could simultaneously test eight groups of diverse samples at six distinct concentrations.

Tests of antioxidant enzyme expression, inhibition of pro-oxidant enzymes, and activation vs. repression of redox transcription factors are also included in the evaluation of antioxidant activity at the cellular level [15]. These tests are in addition to the ability to scavenge ROS/RNS. Caco-2 cells were used to test the antioxidant properties of the extracts made from five brown seaweeds. Both the activity of the antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD) and the amount of glutathione (GSH) present were evaluated [24]. According to these cellular assays, *Pelvetia canaliculata* could exert its antioxidant capacity in Caco-2 cells primarily by preventing H2O2-mediated SOD depletion. In addition, the antioxidant enzyme activities of glutathione peroxidase (GPx) and glutathione reductase (GR) in three Argentine red wines were evaluated. Wine was found to have some protective effects on H2O2 exposed cells, which were attributed to the increased activity of the antioxidant enzymes GPx and GR. In addition, phenols (like curcumin) or food extracts (like blueberries) have been used to treat cultured cells, resulting in a suppression of NF-B

activation as an anti-oxidant response. Curcumin treatment reduced NF-B and activator protein-1 activation as well as IL-8 release in alveolar epithelial cells, according to a study. GSH levels and mRNA expression of the glutamylcysteine ligase catalytic subunit were also higher in treated cells than in untreated ones [25–27].

The ability of antioxidants to slow down the oxidation of 2,2′-azobis-2-methylpropanimidamide, dihydrochloride, (AAPH) or 2,2′-azobis(2-amidinopropane) dihydrochloride, dihydrochloride, (ABAP) is measured using the total radical trapping antioxidant potential (TRAP) assay [28–30]. The variation in the rate of the reaction is measured using fluorometry (ex = 495 nm and e When compared to the rate before the antioxidants were added, the reaction's rate of fluorescence decay slows after they were added [31, 32]. The lag phase duration in comparison to Trolox's lag phase serves as the basis for the quantification. The assumption that antioxidants exhibit a lag phase and that the length of the lag phase is positively correlated with antioxidant capacity underpins the use of the lag phase. However, the potential of antioxidants that play a role after the lag phase is completely ignored because not every antioxidant component possesses an obvious lag phase [33–35].

As a more physiologically relevant measure of antioxidant capacity, the inhibition of induced lipid autoxidation has been developed. Free radical initiator (Cu(II) or 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH)), substrate (linoleic acid or LDL), and antioxidants are typically present in the reaction solution. Cu(II) or AAPH causes linoleic acid autoxidation, or LDL. A UV spectrometer measures conjugated dienes' peroxidation at 234 nm for the lipid components. The reaction begins when a radical initiator is present, and the accumulation of conjugated diene oxides is indicated by an increase in absorbance at 234 nm. The reaction rate slows down after antioxidants are added until the antioxidant is used up. The lag time is measured during the period and used to evaluate antioxidant capacity [36, 37].

The use of a biologically relevant substrate, which makes the results relevant to oxidative reactions in vivo, is this method's main advantage over other in vitro assays. One of the major drawbacks of this method is the variability of the LDL samples, which can vary between donors because LDL is isolated from blood samples. As a result, it is challenging to develop this approach into a high-throughput antioxidant evaluation assay that is consistent and repeatable. The results, on the other hand, would be more reproducible if linoleic acid or its methyl ester was used as an oxidation substrate rather than LDL. However, in the presence of water, linoleic acid would form micelles, and since UV absorbance cannot directly monitor the progression of the reaction in micelles, the method's accuracy may be compromised [38–40].

### **2.1 Natural sources of polyphenols**

Polyphenols, such as phenolic acids, flavonoids, lignans, and stilbenes, are found in a lot of food and medicinal plants. Examples of phenolic acids include cinnamic acid derivatives like p-coumaric, caffeic, and ferulic, as well as benzoic acid derivatives like gallic acid and hydroxybenzoic acids. The hydroxycinnamic acids are more prevalent in edible plants than the hydroxybenzoic acids [41, 42]. The hydroxycinnamic acids are found to be abundant in fruits like blueberries, kiwis, plums, cherries, and apples (0.5–2 g hydroxycinnamic acids/kg fresh wt). While ferulic acid is the most abundant phenolic acid in cereal grains and accounts for approximately 90% of the total polyphenol content of wheat grain, caffeineic acid is the most abundant phenolic acid and accounts for 75–100% of the total hydroxycinnamic acid content in

many fruits. Except for certain red fruits, black radish, and onions, edible plants typically contain very little hydroxybenzoic acid. They are not thought to be particularly nutritious due to their low content [43–45].

The majority of edible fruits and vegetables contain a lot of flavonoids. Flavonols, flavanones, catechins, flavones, anthocyanidins, and isoflavonoids are among its subclasses. Flavonoids come in a variety of forms and concentrations from various food sources. In edible plants, quercetin is typically the most abundant flavonol. Onion is the food with the most quercetin in it. Quercetin levels are relatively low in wine and tea. Kaempferol (broccoli), myricetin (berries), and isorhamnetin (onions) are additional flavonols. Citrus fruits are almost entirely devoid of flavanones. Oranges and mandarins contain the most hesperidin and narirutin flavonoids, while grapefruit contains the most naringin and narirutin flavonoids. Catechins as a rule exist as aglycones or esterified with gallic corrosive. Tea and red wine are the two foods that contain the most catechins [46–48]. Additionally, luteolin and apigenin are the most important flavones. Celery and red pepper are the primary sources for the diet. Anthocyanins like pelargonidin, cyanidin, and delphinidin are what give edible plants like plums, eggplant, and many berries their red, blue, or violet hues [49–51]. The isoflavonoids, for example, isoflavones genistein and daidzein, principally exist in vegetables. Soybean and soy products are the most abundant food source [52, 53].

Linseed, which contains low amounts of matairesinol and secoisolariciresinol (up to 3.7 g/kg dry wt), is the most abundant dietary source of lignans. These same lignans are also found in other algae, leguminous plants like lentils, cereals like wheat and triticale, fruit like pears and prunes, and certain vegetables like garlic, asparagus, and carrots. Resveratrol is a stilbene whose numerous bioactivities have been extensively studied. Resveratrol (0.3–7 mg aglycones/L and 15 mg glycosides/L) is abundant in red wine [54–56].

### **2.2 Natural sources of carotenoids**

Natural pigments called carotenoids include -carotene, lycopene, lutein, and zeaxanthin. All beautiful palatable plants, particularly dim green and yellow-orange verdant, are the great wellsprings of carotenoids [57]. Carotenoids' absorption is primarily dependent on their preparation with oils or fats due to thesir lipid solubility. Among the carotenoids, −carotene is most frequently found in edible plants with the highest provitamin A activity, like acerola, mango, carrot, nuts, and oil palm [58, 59]. A type of red pigment is called lycopene. It almost only exists in the tissues of algae and vegetables. Tomato items like juices, soups, sauces, and ketchup, as well as their handling waste and strip are significant wellsprings of lycopene. The trans isomer accounts for the majority of the lycopene found in tomatoes (between 79 and 91%) [60–62]. The most prevalent xanthophylls found in green and dark leafy vegetables like lettuce, spinach, peas, and broccoli are lutein and zeaxanthin. Zeaxanthin, which accounts for 97.4% of all carotenoids, is also found in the red marine microalga *P. cruentum*.

### **3. Conclusion**

In conclusion, the various nutritional functions and health benefits of antioxidants derived from food and medicinal plants have been the subject of increasing research. Natural antioxidant extraction and antioxidant activity assessment techniques, as well

### *Antioxidants Sources DOI: http://dx.doi.org/10.5772/intechopen.110659*

as their primary sources from food and medicinal plants, are summarized in this review [63–65]. Due to their reduced extraction time, energy consumption, and use of harmful organic solvents, as well as their higher extraction yields for recovering antioxidant compounds from food and medicinal plants, the aforementioned non-conventional methods have the potential to replace or enhance existing extraction methods [66]. Despite this, the majority of them are not suitable for use in industrial settings due to the complicated installation procedures and high cost of the equipment. As a result, finding a balance between cost and energy will be a crucial area of study in the future. The future development trend would be the combination of multiple extraction technologies and the automated potential of these non-conventional extraction technologies to take advantage of the various extraction methods and minimize their disadvantages [67]. The determination of total polyphenolic content by FCR, scavenging free radical ability by TEAC, metal-reducing activity by FRAP, and a kind of cellular-based assay are all suggested for assessing the antioxidant activity of plant materials. Standardizing the operating conditions of the same analysis method and the expression of results is also recommended to make it possible to compare various samples and studies [68–70].

### **Author details**

Marjan Assefi1 \*, Kai-Uwe Lewandrowski2 , Sohila Nankali3 and Alireza Sharafshah4


\*Address all correspondence to: massefi@aggies.ncat.edu

© 2023 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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### **Chapter 2**

## Dietary Regulation of Keap1/Nrf2/ARE Pathway: Focus on Acai Berries and Pistachios and Cashews as Natural Food Sources

*Rosanna Di Paola, Salvatore Cuzzocrea, Roberta Fusco and Marika Cordaro*

### **Abstract**

Inflammation is a biological reaction to oxidative stress in which cell starts producing proteins, enzymes, and other substances to restore homeostasis, while oxidative stress could be intrinsically a biochemical imbalance of the physiologically redox status of the intracellular environment. The nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway, which controls the transcription of numerous antioxidant genes that protect cellular homeostasis and detoxification genes that process and eliminate all toxic compounds and substances before they can cause damage. The Nrf2 pathway is the heart of the daily biological response to oxidative stress. Transient activation of Nrf2 by diet can upregulate antioxidant enzymes to protect cells against oxidative stress inducers. In this chapter, we summarize the effects of some novel foods in the regulation of the Nrf2/ARE pathway and its cellular mechanisms.

**Keywords:** food, oxidative stress, inflammation, diet, Nrf2

### **1. Introduction**

A diet rich in fruits and vegetables has numerous positive effects on the body. In fact, in recent years, research has turned its attention to substances of natural origin: these are rich in essential nutrients with potential therapeutic actions. Nutrients include mainly: vitamins, minerals, fiber, fatty acids, flavonoids, anthocyanins, and carotenoids; the presence of these mainly gives it antioxidant, anti-inflammatory, antimicrobial, antiproliferative, hypoglycemic, cholesterol-lowering, neuroprotective, and cardioprotective action [1]. Recently, the consumption of dried fruits and by-products has gained special attention, among them we can mention in these chapter Cashews, Acai berries, and Pistachios. These components give it anti-inflammatory, antioxidant, antimicrobial, antiproliferative, and astringent actions thanks to the presence of nutrients and substances with different therapeutic actions that give them mainly action against inflammation and oxidative stress as demonstrated in several studies both in vivo and in vitro. Additionally, we briefly discuss the two main molecular pathways involved: NF-E2-related factor 2 (Nrf2) for oxidative stress and NFkB for inflammation (**Figure 1**).

### **1.1 Nrf2**

Nrf2 is one of the most important regulators that shields cells from ROS and xenobiotics that play a key role against the production of antioxidant and detoxifying enzymes [2, 3]. Nrf2 shields cells against stressors such as xenobiotics in food, radiation, reactive oxygen species (ROS), and endogenous chemicals. As a result, activating the Nrf2 pathway may be a viable chemoprevention method [4]. ROS act as a second messenger in cellular communication, but they can alter natural components as lipids, proteins, and DNA, having a detrimental effect on the biological system [5]. Nrf2 is a member of basic leucine zipper genes (bZIP) that are universally expressed in a variety of tissues and cell types and have a conserved structural domain known as a cap'n'collar domain. The leucine zipper region basic's portion is in charge of DNA binding, whereas the acidic area is necessary for transcriptional activation. The heterodimerization of Nrf2 with other bZIP proteins is required for ARE-mediated transcriptional activation [6]. Keap1, an E3 ubiquitin ligase substrate adaptor that is redox-sensitive, controls how much Nrf2 is present inside the cell [7]. Keap1 interacts to Nrf2 in the cytoplasm when the body is not under stress, promoting ubiquitination and proteasomal destruction of Nrf2. The ubiquitin E3 ligase activity of the Keap1-Cul3 complex decreases with exposure to chemicals (typically electrophiles) or ROS, and Nrf2 is stabilized. As it builds up in the nucleus, stable Nrf2 activates the target genes [8]. Under oxidative stress, free freshly produced Nrf2 translocates to the nucleus and heterodimerizes with one of the small Maf (musculoaponeurotic fibrosarcoma oncogene homolog) proteins. The Nrf2-Keap1 association is resolved in a dose-dependent manner. The enhancer sequences known as antioxidant response elements (AREs), which are found in the regulatory regions of Nrf2 target genes. Nrf2 coordinates the expression of several genes, including not only genes encoding antioxidant enzymes but also a series of genes involved in various processes

### *Dietary Regulation of Keap1/Nrf2/ARE Pathway: Focus on Acai Berries and Pistachios and… DOI: http://dx.doi.org/10.5772/intechopen.109239*

including respiratory, cerebrovascular, and neurodegenerative diseases [9–11]. In **Figure 2**, the mechanism of action of Nrf2 is clearly demonstrated. Briefly, (1) Nrf2 is sequestered to the cytoplasm through binding with Keap1 and continually shuttled to the proteasome for degradation. (2) After a response to external stressors, Keap1 cysteine residues are oxidized and Nrf2 serine (Ser) 40 is phosphorylated by protein kinase C (PKC). (3) Nrf2 is then able to translocate into the nucleus and bind to ARE responsive genes in order to increase or decrease their expression. (4) Subsequently, a delayed response to external stressors causes the phosphorylation of GSK-3β by tyrosine (Tyr) kinases. (5) GSK-3β then activates Src kinases, allowing for their translocation into the nucleus. (6) These Src kinases phosphorylate Nrf2 Tyr568, which allows for nuclear export, (7) ubiquitination, and degradation of Nrf2. (8) However, if the insulin receptor signaling is initiated, GSK-3β activity is inhibited. (9) Keap1 is also able to regulate Nrf2 activity through sequestration with PGAM5 to the mitochondria [12].

Multiple genes are impacted by Nrf2 that encode proteins serving as redox balancing agents, detoxifying enzymes, stress response proteins, and metabolic enzymes [6]. Examples of antioxidant detoxification enzymes induced by Nrf2 include heme oxygenase 1 (HO-1) and manganese-dependent superoxide dismutase (Mn-SOD) [13]. Nuclear HO-1 interacts with Nrf2 under oxidative stress, preventing GSK3 mediated phosphorylation along with ubiquitin-proteasomal destruction and extending its accumulation in the nucleus. The preferential transcription of phase II detoxifying enzymes such NQO1 and glucose-6-phosphate dehydrogenase (G6PDH), a regulator of the pentose phosphate pathway, depends on this control of Nrf2 post-induction by nuclear HO-1 [14]. Moreover, the SODs are a family of antioxidant enzymes that catalyze the dismutation of superoxide free radical anions, which are generated during a variety of metabolic activities and lead to the creation of oxygen and hydrogen peroxide molecules. Copper-zinc SOD (Cu, Zn-SOD) and MnSOD, the two primary forms of SODs, are located in the cytoplasm and mitochondria, respectively [15]. It was demonstrated that Nrf2-mediated upregulation of antioxidant

**Figure 2.** *Schematic diagram of Nrf2 regulation.*


### **Table 1.**

*Selected Nrf2 activators present in diet.*

enzymes as GSTs and MnSOD would act to minimize oxidative-stress-induced damage [16].

### **1.2 Nrf2 and NF-ĸB**

To maintain the physiological balance of cellular redox state and to control the cellular response to stress and inflammation, it is hypothesized that Nrf2 and NF-ĸB signaling pathways work in concert. NF-ĸB is a complex protein system constituted by transcription factors that regulate the expression of genes influencing innate and adaptive immunity, inflammation, oxidative stress responses, and B-cell development. NF-ĸB proteins can be divided into two classes according to whether they include or lack a transactivation domain. Since p50 and p52 lack the transactivation domains that RelA (p65), RelB, and c-Rel possess. Heterodimerization with the Rel proteins is necessary for them to activate transcription [17]. Nrf2/ARE signaling plays a crucial role in the protection against oxidative stress and is responsible for the maintenance of homeostasis and redox balance in cells and tissues. In contrast, NF-ĸB is also a redox-regulated transcription factor, which regulates inflammatory responses and cellular injury [18]. Firstly, Nrf2 inhibits oxidative-stress-mediated NF-κB activation by decreasing the intracellular ROS levels. Furthermore, Nrf2 prevents the IκB-α proteasomal degradation and inhibits nuclear translocation of NF-κB [19]. Studies suggest that Nrf2 counteracts the NF-ĸB-driven inflammatory response by competing with transcription co-activator cAMP response element (CREB) binding protein (CBP) [20, 21]. Histones are acetylated by the CBP-p300 complex, which also makes DNA accessible for the construction of the transcriptional machinery.

*Dietary Regulation of Keap1/Nrf2/ARE Pathway: Focus on Acai Berries and Pistachios and… DOI: http://dx.doi.org/10.5772/intechopen.109239*

Additionally, the Nrf2 and p65 non-histone proteins, as well as others, have their lysine residues acetylated by the CBP-p300 complex. Since, CBP also preferentially interacts with p65, the overexpression of p65 limits the availability of CBP for Nrf2 interaction; accordingly, knockdown of p65 promotes Nrf2 complex formation with CBP (**Table 1**) [38].

### **2. Açai berry**

The Açaí berry is a little, spherical fruit (about the size of a grape) that is green while immature and turns dark purple when it is fully developed. It comes from the Açaí palm, a native of Central and South America that also thrives in marshes and flood plains in addition to the Amazon region. Açaí berries are eaten fresh or juiced as food. The juice can be used as a natural food colorant and is commercially employed in jelly, syrup, ice cream, liquors, energy drinks, and a range of other beverages [39]. Açaí juice is viscous and contains 5.9% fats and 2.4% protein. The apple pulp has 12% fats and 4% protein. Vitamins A, C, and E, calcium, phosphorus, iron, and thiamine are among the nutrients. The Açaí berries of the *Euterpe oleracea* plant are thought to be a source of bioactive substances, particularly anthocyanins and unsaturated fatty acids, which are known to have health-promoting properties. These berries may help to reduce metabolic stress and inflammation while enhancing antioxidant protection. Orientin, isoorientin, vanillic acid, as well as the anthocyanins cyanidin-3-glucoside and cyanidin-3-rutinoside, are only a few of the polyphenolic components having antioxidant capabilities found in acai extracts. Acai pulp is rich in proanthocyanins and total phenolics, but also contains trace amounts of anthocyanins. Industrially processed samples have a significant percentage of proanthocyanidins, but naturally occurring anthocyanins are significantly enriched (20 times more). The unprocessed Açai pulp extracts reduced the expression of pro-inflammatory genes such as interleukin-1, cyclooxygenase-2, nitric oxide synthase, and interleukin-6 and dramatically inhibited the generation of nitric oxide, which has been linked to proanthocyanidins in the initial inflammatory response [40]. These chemicals' existence is mostly associated with their anti-inflammatory, antiproliferative, antioxidant, and cardioprotective properties [41]. Açai can be used to treat certain diseases due to its anti-inflammatory and antioxidant effects, acting at the level of the Nrf2 pathway. It has also been shown that Açai may act on different pathways; for example, according to some studies, it goes to act on peroxisome proliferator-activated receptors (PPARs) α and γ, going to decrease the transcription of various genes, including pro-inflammatory, prooxidants, and those affecting lipid metabolism genes. Another pathway that goes to modulate is that of Nf-kB, going to decrease the production of pro-inflammatory cytokines. For example, the fruit has been used to treat Vascular Dementia (VaD), which is the secondary most frequent reason for inherited biological cognitive impairment [42]. VaD is caused by an oxidative stress increase, which might cause cognitive decline brought on by aging and neurological diseases. Since oxidative and inflammatory stressors may reduce synaptic plasticity and memory by resulting in dendritic modification and cell death, important brain regions such as the hippocampus should be more susceptible to these situations. Additionally, the expression of microtubule-associated protein 2 (MAP-2) and α-Tubulin, two significant neuronal markers of well-being, is altered in the brain of VaD patients [43, 44]. Furthermore, it has been demonstrated that impaired autophagy alters protein

"quality control," accumulates unwanted proteins and organelles in brain cells [45]. In this study, Açai Berry was useful to conteract VaD alterations in the brain. Açaí berries can also mitigate Parkinson's disease progression, which is the second most prevalent neurological condition in people over 65 [46]. Recent studies have revealed that the Nrf2/ARE signaling cascade is the most likely target for therapeutic therapy, despite advancements in our understanding of the pathophysiology of PD [47]. The oral administration of Açaí berries has shown an important decrease of ROS and an increase of Nrf2 expression. Also, in this study, the authors observed a significantly improvement in both motor and non-motor deficits, histological alteration, pro-inflammatory cytokine release, neutrophilic infiltration, and lipid peroxidation limiting dopaminergic neuronal death [48]. Some studies, additionally, have shown Açai berries' anti-inflammatory properties in a model of co-culture between Caco-2 and RAW 264.7 macrophages, which has the potential to prevent intestinal inflammatory diseases, due to anthocyanin improved Tight Junction barrier integrity and reduce gastrointestinal inflammation by preventing the expression of cytokines, especially IL-6, IL-8, and PGE2 through inhibition of COX-2 [49]. Açai barriers can also be used for anti-inflammatory treatment of bone diseases such as periodontal disease are triggered by chronic inflammation causing the upregulation of osteoclastogenesis. This in turn shifts the bone remodeling process toward increased bone resorption [50]. Inflammatory cytokines have been associated with bone destruction, having a role in the regulation of the expression of the receptor activator of nuclear factor kappa B (RANK) and receptor activator of nuclear factor kappa B ligand (RANKL), which is a vital step in the activation of osteoclastogenesis [51]. Açai-berry extract (ABE) on the reduction of osteoclast formation and resorptive activity of RANKL-induced osteoclast precursor cells. Moreover, ABE also modulated the secretion of several inflammatory cytokines during osteoclastogenesis and osteoclast activity [52]. Açai seeds, according to some studies, regulate NF-κB and Nrf2/ARE pathways protecting lung against acute and chronic inflammation [53]. Moreover, studies have shown the cytotoxic effect of Açai seeds against the MCF-7 breast cancer cell line. This effect is given by the ability to induce ROS synthesis inside these cells. But also, it induces morphological changes and reduces cells viability, due to flavonoids content [54]. In high-fat mice, Açai seed extract reduces the activation of the renin-angiotensin system, oxidative stress, and inflammation in the white adipose tissue [55]. Acai seed prevented the body weight rise brought on by the heart failure diet, which was correlated with the diminution of adipocyte area and the accumulation of visceral fat, indicating that the diminution of adipose mass may contribute to the acai seed-mediated decrease in body weight. These advantageous effects of acai seed were also connected to a significant decrease in the serum levels of total cholesterol (TC), triglycerides (TG), very low-density lipoprotein (VLDL), and low-density lipoprotein (LDL), indicating a favorable impact of Açai seed on the altered lipid profile [56]. The pulp of Açai can be used, for example, to mitigate colitis-associated colon carcinogenesis, which is one of the most common cancers in the modern world. A lesion caused by acute inflammation was characterized microscopically by a coagulative necrosis process and had macroscopic signs of necrosis. According to the phytochemical tests, the lyophilized acai pulp (AP) utilized in the in vivo trial included significant amounts of the phytonutrients cyanidin 3-rutinoside (C3R) and cyanidin 3-glusoside (C3G). Additionally, the concentration of anthocyanins may range between açai samples utilized in various research. Cells' ability to move slightly less after receiving acai pulp treatment [57, 58].

*Dietary Regulation of Keap1/Nrf2/ARE Pathway: Focus on Acai Berries and Pistachios and… DOI: http://dx.doi.org/10.5772/intechopen.109239*

### **3. Pistachios**

Pistachios originate in West Asia and are traded in the Mediterranean, Europe, and the East. The only species that produces edible nuts is *Pistacia Vera* L. (Pistacio), which is a member of the Anacardiaceae family [59]. The fact that pistachio plants can grow in a variety of soil types and survive dryness is crucial for sustainability because semi-arid regions require vital water consumption. The pistachio fruit is an edible drupe with a thin, soft coating. The endocarp, which is covered with a fleshy, thin hull that is light green in color with red undertones, is inedible. In comparison to other nuts, pistachios contain a high concentration of compounds that have antioxidant and anti-inflammatory properties [60]. Nuts have positive health effects on a variety of metabolic conditions, including hypercholesterolemia, hyperglycemia, hyperhomocysteinemia, and everything else that goes along with them. Pistachios has a high nutritional content and is consumed frequently over the world because it has significant nutritional properties and offers many health advantages [60]. One of the foods that must be included in a nutritious and balanced diet is the eating of nuts. Protein, fiber, monounsaturated fatty acids, minerals, and vitamins are all present in excellent amounts in pistachios, but they are also a good source of carotenoids, phenolic acids, flavonoids, and anthocyanins (**Table 2**).

Lutein, zeaxanthin, and a variety of other bioactive phenolic compounds found in pistachios help to improve endothelial function, glycemic management, and antioxidant and anti-inflammatory activity. The highest concentrations of potassium, tocoferol, and phytosteroids can be found in citrus fruits [61, 62]. Lipophilic extracts from the peel and kernel of raw shelled pistachios contain fatty acids, phytosterols, and tocopherols, according to phytochemical study. These polyphenols in pistachios have strong antioxidant action. Gallic acid and other phenolic chemicals, such as phenol acids, flavonoids, stylibenes, and tannins, have one or more aromatic rings and hydroxyl groups [63–65]. As a great source of phenolic compounds, pistachios have strong antioxidant properties that can block ROS, preventing the oxidation of biological macromolecules [66]. The activity of the various pistachio nut components was evaluated in a number of in vitro and in vivo investigations, and the various lipophilic (carotenoids, tocopherols, and chlorophyll) and hydrophilic extracts were


### **Table 2.**

*Macronutrients content in 100 g of Pistacio. Source: U.S. Department of Agriculture Food Data Central 2019.*

compared [67]. The hydrophilic extract exhibits higher antioxidant activity than the lipophilic components in the kernel, and this activity has been observed to block the metal-dependent and independent lipid oxidation of bovine liver microsomes in a dose-dependent manner [68]. Human low-density lipoprotein (LDL) has also been shown to oxidize less when exposed to copper [60]. Compared with the kernel, the tegument of the pistachio contains a higher level of antioxidant activity. By combining lipophilic and hydrophilic extracts with macrophages that have been stimulated by lipopolysaccharide (LPS), this was proven [69]. The hydrophilic tegument extracts shows stronger inhibition by subsequently reducing nitric oxide (NO) production. The extracts markedly decreased ROS formation. According to the findings of this in vitro study, the tegument extract had a higher concentration of phenolic compounds and hence had more antioxidant activity. In mature adipocytes, these fractions greatly decreased lipid accumulation. Additionally, it has been proposed that the antiproliferative properties of pistachios contribute to their anticancer properties. The growth of LT97 colon adenoma cells has been shown to be inhibited by pistachio fermentation supernatants in vitro in a dose-dependent manner [66]. Additionally, pistachio fermentation supernatants have been shown to increase antioxidant activity, which promotes the expression of catalase (CAT), which lessens DNA damage brought on by hydrogen peroxide (22) [70]. According to the findings of these investigations, roasting pistachios may alter their phytochemical composition and improve biological activity [71]. The gut microbiota, a complex ecology that varies according to anatomical location, is another crucial area of study in science. Obesity, type 2 diabetes, and other illnesses can sometimes cause the microbiota to become out of balance and enter a state of dysbiosis [72, 73]. Diet also plays a significant part in this. According to a study comparing the intake of almonds and pistachios on treated volunteers, the consumption of pistachios was able to change the microbiota's composition more than almonds [74]. According to studies on the microbiome, eating pistachios in moderation can help the body's microbiota get back into balance by boosting the population of helpful bacteria and lowering acute inflammatory conditions. In fact, pistachio supplementation has been found to repair the intestinal microbiota in diabetic rats on a high-fat diet. Drug resistance is a widespread issue, and novel treatments are the focus of current research. Because they include bioactive substances that can be employed as antimicrobials and antivirals, plant extracts play a significant role in medicine. Bactericide properties of raw, salted, roasted pistachios have been demonstrated. Additionally, the effectiveness of a Pistacia Vera metabolic extract against staphylococcal infections has been demonstrated. Pistachios contain polyphenols, which can be extracted alone or combined with other medications to make a potent alternative to antibiotics [75]. Additionally, polyphenols have antiviral properties. Pistachios contain polyphenols, which can be extracted alone or combined with other medications to make a potent alternative to antibiotics. Additionally, polyphenols have antiviral properties. This has been shown to prevent replication of Herpes Simplex Virus Type 1 (HSV-1). Pure polyphenol extracts were used to treat the condition, which inhibited the expression of many viral proteins and the creation of viral DNA [76]. It is important to keep in mind that pistachio component quantities can differ depending on genotype, pre- and post-harvest circumstances, and storage [77]. Numerous experimental models have been used to examine the anti-inflammatory properties of pistachio components in acute inflammatory states such paw edema [78–81], LPS inflammation [69], and chronic inflammation models such as colitis [82]. By contrasting raw, shelled pistachios with salted and roasted pistachios, the therapeutic effects of pistachios were discovered in an experimental animal model of

### *Dietary Regulation of Keap1/Nrf2/ARE Pathway: Focus on Acai Berries and Pistachios and… DOI: http://dx.doi.org/10.5772/intechopen.109239*

paw edema generated in rats. In contrast to roasting, which results in a 60% drop-in antioxidant activity, eating raw shelled natural pistachios has been shown to result in reduced nitrate protein production [68, 82, 83]. A diet with a balanced intake of pistachios has been demonstrated to enhance serum concentrations of tocopherol, lutein, and carotene. In addition, pistachio consumption has been proven to decrease oxidized LDL concentrations in randomized trials of healthy patients and hypercholesterolemic subjects [84]. Malondialdehyde (MDA), a by-product of lipid peroxidation, was reduced, and blood antioxidant potential was improved by eating pistachios [85]. Numerous studies have demonstrated the critical role played by bioactive components in mastic oil produced from Pistachio Lentiscus in the treatment of ulcerative colitis, where inflammation and oxidative stress play a significant role. Myeloperoxidase (MPO) activity was dramatically decreased by flavonoids and other bioactive substances [86]. Mastic oil therapy reduces the inflammatory response of ulcerative colitis, which is mediated by cytokines such as TNF- and IL-6. These research studies sought to emphasize the critical function of the pistachio's bioactive components and the potential significance of including them in a nutritious, wellbalanced diet [87, 88]. In particular, Nrf2 pathway plays a significant role in antioxidant activity. When there is a redox imbalance, this pathway becomes less active, which depletes the body's supply of antioxidant enzymes. The release of pro-inflammatory cytokines can also activate the NF-B signaling pathway, which results in decreased Nrf2 pathway activity and oxidative stress conditions. Inflammatory response and oxidative stress are modulated, according to a study done after the extraction of polysaccharides from *Pistacia vera* L. Pistachio polysaccharides decreased inflammation and oxidative stress by boosting antioxidant production through the Nrf2 pathway and attenuating the NF-kB pathway [89, 90]. The positive effects of bioactive substances are dose-dependent, it should be noted. Additionally, research has been done on the therapeutic effects of pistachios in experimental models of neurodegenerative diseases for cognitive problems [91]. In particular, lutein and zeaxanthin improved cellular communication required for light processing and the growth of neural circuits in the visual system, which helped improve memory and motor performance when pistachios were supplemented [92]. The anti-inflammatory and antioxidant properties of pistachio bioactive components are dose-dependent, it should be noted. By preventing the cellular aging phenomenon brought on by inflammation and oxidative stress, their balanced consumption in the diet might enhance quality of life.

### **4. Cashew**

Cashew (*Anacardium occidentale L*.) is a tree that originates in Brazil, but with the exploration has also spread to Asia and Africa. Cashew is a perennial plant belonging to the family of the Anacardiaceae: these plants have a considerable height, the trunk is irregular and short, the leaves evergreen elliptical oblate, while flowers are small with sepals and petals gathered in a panicle. The cashew fruit consists of an accessory fruit and a true fruit. The accessory fruit is called cashew apple and, when it reaches full maturity, is a kidney-shaped drupe, inside of which is the cashew nut surrounded by a double shell. From the cultivation of this plant, the fruit is used: both cashew apple and cashew nut; but more recently also a by-product, the cashew nut shell liquid [93–95]. The cashew nut has remarkable nutritional properties due to its various components: lipids including polyunsaturated and monounsaturated

fatty acids; amino acids including glutamic acid, aspartic acid and leucine; minerals such as calcium, potassium, and magnesium. Cashew apple is rich in sugars and minerals; the cashew nut shell liquid is rich in phenols and has antimicrobial properties. In addition, cashew by-products have several properties: the cashew skin extract is a good antioxidant; instead, the bark is rich in tannins and has astringent, anti-inflammatory, hypoglycemic, antibacterial, and antimutagenic properties. Also in cashews other molecules with possible therapeutic effects are: saponins, catenins, tannins, carotenoids, and anthocyanins. With regard to the therapeutic applications of cashew, several studies have shown that its molecules grant it different actions reason why different therapeutic effects are being evaluated recently. Foremost among them, the anacardic acid is an antimicrobical agent, particularly it acts against Gram-positive bacteria; another application of anacardic acid is such as antitumoral agent, in fact has been shown to act by blocking the HAT enzyme: this is an enzyme involved in the acetylation of histones [96]. Furthermore, some studies have highlighted how anacardic acid induces autophagy and apoptosis. Certainly among the most studied therapeutic applications today are the antioxidant and antiinflammatory effects of cashew nuts [97]. The anti-inflammatory effects of cashew are due to its interaction with the transcriptional factor NFkB, specifically there is inhibition of this pathway, resulting in a decrease in proinflammatory cytokines. Antioxidant effects are due to action on several factors: the anacardic acid acts on lipid peroxidation and lipoxygenase [98]; in addition, polyphenols and flavonoids have antioxidant action as they modulate oxidative balance and also have been seen to act on the Nrf2 pathway, promoting its translocation into the nucleus resulting in the synthesis of cytoprotective enzymes such as NADPH quinone dehydrogenase NQO-1 and HO-1. Additionally, the modulation of Nrf2 pathway influences other important molecular pathways and mechanism: such as NLRP3 and apoptosis. Several studies have been conducted in recent years to evaluate the effects of cashew nut consumption, and these have yielded positive effects in the treatment of various diseases. In in vitro studies, the activity of cashew nuts on the human microbiota was evaluated: the effects obtained induce a change in metabolic activity with potential prebiotic activities [99]; the cashew apple juice contains gluco- oligosaccharides that promote the growth of the microbiota [99] . In vivo studies included different experimental protocols going to investigate various pathologies. Among neurodegenerative diseases, a study has been done on Parkinson's disease: the pathology has been induced by rotenone in male rats, and then they were treated with anacardic acid; analyses have shown that there is a decrease in oxidative stress, specifically there is modulation of mitochondrial respiration and superoxide dismutase [100]. In in vivo study was evaluated two aspects of cashew nuts: antioxidant and anti-inflammatory action in different model of inflammatory disease: such as colitis, edema, and pancreatitis. In colitis model, the pathology was induced intrarectally through injection of dinitrobenzene sulfonic acid (DNBS); subsequently, the cashews were administrated orally. Findings have shown that cashew nut consumption inhibits the inflammatory pathway NFkB and activates the expression of antioxidants such as superoxide dismutase [97]. The pancreatitis was induced in CD1 mice by cerulein; in this case, the cashew acts on Nrf2 pathway and on NRLP3 pathway: Nrf2 translocates into the nucleus by inducing the synthesis of cytoprotective enzymes such as HO-1 hemeoxygenase and superoxide dismutase; instead the cashew acts on NRLP3 reducing its levels resulting in decreased pro-inflammatory cytokines. These effects are probably due to its components such as flavonoids and polyphenols [101]. The edema was induced by

### *Dietary Regulation of Keap1/Nrf2/ARE Pathway: Focus on Acai Berries and Pistachios and… DOI: http://dx.doi.org/10.5772/intechopen.109239*

carrageenan injection in male rats, and the results showed that the administration of cashew reduced edema formation and induced the endogenous antioxidants activity; and also in this study is shown the analgesic effect [102]. The antioxidant and antiinflammatory action of cashew nuts is evaluated also in multi-organ pathology such as Hyperhomocysteinemia [103, 104]. Among the cardiovascular diseases considered was ischemia/reperfusion injury, in this model demonstrated that the consumption of cashew acts on lipid peroxidation, tissue myeloperoxidase activity, and reactive oxygen species generation: inducing a decrease of levels; also there is a decrease of pro-inflammatory cytokines and an increase of antioxidant activity. In addition, studies have shown that cashew consumption reduces the risk of cardiovascular disease: probably because of its concentration of fatty acids, which also have a hypolipidemic action [105, 106]. Other studies have shown that not only cashew nut consumption has therapeutic effects but also its derivatives such as extracts. The leaf extract of Anacardium has an anti-inflammatory and bronchodilatory action: this is shown by in vivo studies on animal model. In this case, the effects are due to a derivative present in the extract, oleamide [107]. In the dermatological field, there is some evidence to suggest a possible application of cashews as a dermatological treatment as well, but extensive studies have not yet been conducted [108]. Certainly the in vitro and in vivo studies carried out demonstrate action on inflammation and oxidative stress, particularly cashews modulate important pathways, such as Nrf2 and NFkB. The latest studies instead are also focusing on the anticancer effect, and this food might have evaluated antiproliferative action on cancer cells [109].

### **5. Conclusion**

It has been increasingly clear in recent years how nutrition may affect the prevention and/or treatment of several chronic diseases. Based on the food ingredients that can have positive effects on health, a balanced and diverse diet is advised. For instance, antioxidant chemicals can fight free radicals directly or indirectly by boosting cellular endogenous antioxidant defenses, such as by activating Nrf2. Resveratrol, catechin, and allicin are a few chemical substances found in the human diet that have strong biological effects and may be good for cardiovascular health. They also prevent ROS damage by upregulating phase II detoxifying enzymes and raising levels of cellular glutathione. Açai berries, cashew nuts, and pistachios are some of the bioactive ingredients of the diet that are covered in this chapter. Since the majority of studies are in vitro or in animals, and it is unknown how far these doses can be extrapolated to be effective in humans, it is not yet possible to establish safe and effective doses for supplementation, taking into account all the studies that have been discussed in this chapter. The usage of food ingredients does, however, seems to have the benefits of relatively low toxicity, a wealth of resources, and low cost. Therefore, "nutritional therapy" emerges as a crucial method for preventing and/or treating a variety of diseases, enhancing the welfare of people, and trials to determine their efficacy should be carried out.

### **Acknowledgements**

These authors want to thank Doctors Livia Interdonato, Ylenia Marino, Alessia Arangia, and Gianluca Antonio Franco for their incredible contribution.

### **Conflict of interest**

The authors declare no conflict of interest.

### **Author details**

Rosanna Di Paola1 , Salvatore Cuzzocrea1 , Roberta Fusco1 \* and Marika Cordaro2

1 Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Messina, Italy

2 Department of Biomedical, Dental and Morphological and Functional Imaging, University of Messina, Messina, Italy

\*Address all correspondence to: rfusco@unime.it

© 2022 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.

*Dietary Regulation of Keap1/Nrf2/ARE Pathway: Focus on Acai Berries and Pistachios and… DOI: http://dx.doi.org/10.5772/intechopen.109239*

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### **Chapter 3**

## Antioxidants Obtained from the Natural Sources: Importance in Human Health

*Sushil S. Burle, Krishna R. Gupta, Swati N. Lade, Shyam W. Rangari and Milind J. Umekar*

### **Abstract**

Now a day the interest in natural and synthetic antioxidants is increasing very rapidly in functional food ingredients and dietary supplements. The differences between the number of free radicals and antioxidants are the main cause of the oxidative damage of lipids, proteins, and DNA. In this chapter, we are summarising the natural antioxidants which have been obtained from plants, animals, or microbial sources. Flavonoids are the most comprehensive antioxidant compounds which are obtained from natural sources. These flavonoids are reactive toward many radicals which are studied by many researchers under various experimental conditions and their structural activity relationships have been recognised. This chapter includes the various types of antioxidants obtained from natural sources and their impact on human health as pharmaceutical, nutraceutical, and phytoceuticals as well as their use in the treatment of various diseases along with the mechanism of action.

**Keywords:** free radical, natural antioxidants, phytoceuticals

### **1. Introduction**

The antioxidant is defined as the any substance at very low concentration compared with that of an oxidizable substrate significantly which delays or inhibits oxidation of the substrate. According to the Halliwell and Gutteridge the antioxidants are the substance that prevents oxidative damage to a target molecules [1]. The antioxidant act as a oxidation inhibitors at very low concentration when people are using them to prevent the health damages ours due to the polluted plants and various factors which causes the illness in human beings [2]. For the survival of human beings the oxygen plays very important role under some situation it shows the deleterious effects on the human body. The negative effects of oxygen are due to the formation and activity of number of chemical compounds it knows as the reactive oxygen species (ROS). These ROS is collectively including both oxygen radicals and several non-radical oxidising agents that mostly take part in the intitation or propagation of chain reaction [3]. The reactive species are free radicals that represent a class of highly reactive intermediate chemical entities whose reactivity is derived from the presences of unpaired electron in the chemical formula structure. There are the two main major group in the living cells: enzymatic and non enzymatic antioxidants these enzymatic are again further divided into the primary and secondary enzymatic. The primary is composed of three important enzymes which prevents the formation and neutralisation of the free radicals by donating two electrons to reduces the peroxidase by forming selenols and also eliminates peroxidase as potential substrate for the fenton reaction catalase which turns hydrogen peroxide into water and molecular oxygen one of the most important and efficient antioxidants known today which turns hydrogen peroxide into water and molecular oxygen—one of the most important and efficient antioxidants known today, when just one molecule of catalase converts 6 billion molecules of hydrogen peroxide. The superoxide dismutase which changes the superoxide anions into hydrogen peroxide which is act as catalase.

### **1.1 Antioxidant**

The antioxidant is the substances that prevent the oxidative damage in the body all the cell in the body requires the oxygen (O2) for energy production and naturally produced free radical as a byproduct which causes damage. Antioxidants act as "free radical scavengers" and repair the damage done by free radicals. This term was used in late nineteen and early twenty centuries. The antioxidant molecules are capable of preventing the oxidation of other molecules. These are obtained from both naturally as well as synthetic source [4].

### *1.1.1 Types of antioxidant*

Antioxidants are classified in two ways depending on their solubility one is hydrophilic which is soluble in water and other hydrophobic which is water-insoluble but soluble in lipids. The water-soluble antioxidant reacts with oxidant present in cell cytosol and blood plasma. On the other hand, the hydrophobic antioxidant prevents the cell membrane from lipid peroxidation. Traditionally these antioxidants are of two classes' primary or chain-breaking antioxidant and secondary or preventative antioxidant [5, 6].

Mechanisms of primary antioxidant as follows

L• þ AH ! LH þ A• (1)

$$\rm{LO} \bullet + \rm{AH} \rightarrow \rm{LOH} + \rm{A} \bullet \tag{2}$$

$$\text{LCO} \bullet + \text{AH} \rightarrow \text{LCOOH} + \text{A} \bullet \tag{3}$$

$$\mathbf{A}\bullet + \mathbf{A}\bullet \to \mathbf{A}\mathbf{A} \tag{4}$$

Where L• is a lipid radical, AH• inhibited antioxidant (**Figure 1**).

### *1.1.2 Use of Antioxidant*


*Antioxidants Obtained from the Natural Sources: Importance in Human Health DOI: http://dx.doi.org/10.5772/intechopen.109440*

**Figure 1.** *Classification of antioxidants found in natural sources.*


### **2. Free radicals**

These are highly reactive chemical species. These reactive oxygen species generated in phagocytosis, mitochondrial respiratory chain, fertilisation, and arachidonic acid metabolism. These free radicals are a species which contains an unpaired electron in an atomic orbital. They either donate an electron or extract an electron from other molecules. [8] Chain reactions of these radicals are dividing into three parts viz., Initiation, Propagation, and Termination.

In initiation reaction a net increase in the number of free radicals, while in propagation reaction are those reactions involving free radicals in which the total number of free radicals are same. Further in termination reaction, the net reduction in the number of free radicals takes place. Two free radicals combine to form a more stable species.

### **2.1 Types of free radicals**

As a free radical originated from oxygen atom hence it is called as a reactive oxygen species (ROS) this ROS include a superoxide (O2 ), Hydroxyl (OH), hydrogen peroxide (H2O2), peroxyl (ROO), nitric oxide (NO) and alkoxy (RO) until completely reduced to water. The large no superoxides are produced in mitochondrial and microsomal electron chain. On the other hand, the cytochrome oxidase is retained by the moderately reduced oxygen intermediated bound to its active site. All other elements of mitochondrial respiratory chain transfer the electron directly to oxygen and not preserve the reduced oxygen intermediate in the active site [9].

### *2.1.1 Super oxide ion*

This is an oxygen molecule which contains extra electron. This free radical causes damage to DNA, Mitochondria. The body can neutralise the superoxide radical by producing Superoxide dismutase.

### *2.1.2 Hydroxyl radical*

It produced by the reduction of an oxygen molecule in electron transport chain. These hydroxyls radical are highly reactive in nature and cannot eliminate by an enzymatic reaction. As it is more reactive it damages most of the organic molecules like DNA, Lipid, Carbohydrate, and proteins.

### *2.1.3 Nitrogen species*

These are also called reactive nitrogen species. The metals like copper and iron have many numbers of unpaired electrons which can also act as free radicals. These metals have not strong affinity of the electron but can effortlessly accept and donate electrons.

### *2.1.4 Oxygen radical*

The radical are mostly formed by the immune system. These generally cause the oxidation of cholesterol and LDL.

### **2.2 Free radical and biology**

It helps for intracellular killing of bacteria by phagocyte cells such as macrophages and granulocytes.

Superoxide and Hydroxyl radical are most important radical due to their reactivity. These radicals take part in unwanted side reaction this side reaction results in cell damage. As it concentration increase it may lead to cell injury and finally death of the cell. This action is responsible for various disease conditions like myocardial infarction, stroke, cancer, diabetes etc. Few symptoms of ageing like atherosclerosis are also recognised to free radical-induced oxidation of numerous chemicals. Apart from this disease it may also involve in Parkinson's disease, senile deafness caused due to drugs, Alzheimer disease, and schizophrenia [10].

*Antioxidants Obtained from the Natural Sources: Importance in Human Health DOI: http://dx.doi.org/10.5772/intechopen.109440*

### *2.2.1 Sources of free radicals*

The free radicals are the byproduct of cellular processes and these are produced from metal cofactors by spontaneous catalyzation. Nowadays the environmental sources act as a measured source of free radicals as the radiation is increasing largely around the city are also responsible for the radiation sources includes the mobile phone, X-ray, computer and television set etc. apart from this mental stress is also one of the sources of free radical generation [11].

### **3. Oxidation**

The utilisation of oxygen for generation of energy by metabolism of food nutrients is most important part for survival of all living beings. This oxygen is extremely reactive atom which is useful for the destruction of singlet oxygen, hydroperoxyl radical, superoxide radical, organic peroxides, nitric oxide, peroxynitrite and triplet oxygen. The oxygen which is consumed during the breathing it produces free radical production and apart from this environmental factors such as smoke, pollutants, and certain chemicals also contribute to their formation. This leads to the starting of chain reactions in cells and it can cause damage or death to the cell [12]. The Mechanisms with antioxidant properties were purposefully included to fatty meals in order to disrupt chain processes by substituting free radical intermediates and inhibiting other oxidation reactions. Antioxidants, such as ascorbic acid, thols, and polyphenols, help to neutralise free radicals by oxidising themselves and acting as reducing agents. In the packaged food, the antioxidant is added separately to prevent the generation of free radicals and to ensure food safety.

The various properties of radicals like accepting or donating an electron from other molecules which leads to the stabilising the free radical at the beginning but stats to produces another process which leads to the damage of biological molecules like Proteins Carbohydrates lipids and DNA which leads to the homeostatic disruption [13].

### **3.1 Concept of oxidative stress**

The Oxidative stress explain the relation between the disease and free radicals. The normal healthy human body, generates the pro-oxidants in the form of reactive nitrogen species and reactive oxygen species and reactive nitrogen species are useful for the maintains of all antioxidant level. This delicately maintained balance is shifted in favour of pro-oxidants whenever it is exposed to various environmental, physicochemical, and pathological agents such as cigarette smoking, atmospheric pollutants, radiation UV rays, toxic chemicals, overnutrition, and advanced glycation end products (AGEs) in diabetes. It has been linked to the genesis of over 100 human illnesses as well as the ageing process [5].

### **3.2 Molecular damage induced by free radicals**

Various biological molecules present in our body are attacked by the various free radicals which leads to the impairment of cell functions and damaged the molecules produces the diseased states.

### **3.3 Lipids and lipid peroxidation**

The lipids present in membrane are highly susceptible to free radical damage when this lipid reacted with free radicals can undergo highly damaging chain reaction of lipid peroxidation leading to both direct and indirect effects. The lipid peroxidation mediated by the free radical process in this the intitation caused by the species attack which abstract a hydrogen atom from the methylene group which leave an unpaired electron on the carbon atom. Molecular rearrangement stabilises the resulting carbon radical to form a conjugated diene, which can then combine with an oxygen molecule to form a lipid peroxyl radical. These radicals can then extract hydrogen atoms from additional lipid molecules to generate lipid hydroperoxides, further propagating lipid peroxidation. A variety of responses can end the peroxidation process. The most important one involves the reactivity of LOO• or lipid radical (L•) with an antioxidant molecule such as vitamin E or -tocopherol (-TOH), resulting in a more stable tocopherol phenoxyl radical that is not engaged in further chain reactions. Other cellular antioxidants, such as vitamin C or GSH.

Many toxicologically interesting chemicals are produced during the lipid peroxidation process, including malondialdehyde, 4-hydroxynonenal, and other 2-alkenals. Isoprostanes are unique products of arachidonic acid lipid peroxidation, and procedures such as mass spectrometry and ELISA-assay kits have recently become available to identify isoprostanes [14, 15].

### **3.4 Proteins**

Protein oxidation by ROS/RNS can result in the formation of both stable and reactive molecules, such as protein hydroperoxides, which can generate additional radicals when they interact with transition metal ions.

Although the majority of oxidised proteins that are functionally inactive are quickly removed, some can accumulate over time and contribute to the damage associated with ageing as well as a variety of diseases. Lipofuscin, a peroxidized lipid and protein aggregation, forms in the lysosomes of aged cells and Alzheimer's disease brain cells [16].

### **3.5 Carbohydrates**

Many toxicologically interesting chemicals are produced during the lipid peroxidation process, including malondialdehyde, 4-hydroxynonenal, and other 2-alkenals. Isoprostanes are unique products of arachidonic acid lipid peroxidation, and procedures such as mass spectrometry and ELISA-assay kits have recently become available to identify isoprostanes [13].

### **3.6 DNA**

The interaction of DNA with ROS or RNS causes oxidative damage to the DNA. •OH, eaq-, and H• free radicals react with DNA by adding to bases or removing hydrogen atoms from the sugar moiety. •OH attacks the C4dC5 double bond of pyrimidine, resulting in a variety of oxidative pyrimidine damage products such as thymine glycol, uracil glycol, urea residue, 5-hydroxydeoxyuridine, 5-hydroxydeoxycytidine, hydantoin, and others. Similarly, when •OH reacts with purines, it produces 8-hydroxydeoxyguanosine (8-OHdG), 8-hydroxydeoxyadenosine,

formamidopyrimidines, and other unidentified purine oxidative products. Several repair pathways are involved in the repair of DNA damage [9, 17]. 8-OHdG has been linked to cancer and is regarded as a trustworthy marker for oxidative DNA damage.

### **3.7 Significance of antioxidants in relation to disease**

Zinc is a trace element that functions as a cofactor for approximately 200 human enzymes, including the cytoplasmic antioxidant Cu-Zn SOD, an isoenzyme of SOD found mostly in the cytosol. Selenium, a trace element, also serves as a cofactor for glutathione peroxidase. Vitamin E and tocotrienols (produced from palm oil) are powerful lipid-soluble antioxidants that act as a "chain breaker" during lipid peroxidation in cell membranes and other lipid particles like LDL [18, 19].

Vitamin E is considered the "gold standard antioxidant" against which other antioxidant-containing compounds are tested, particularly in terms of biological activity and therapeutic importance. The recommended daily intake varies from 400 to 800 IU. Ascorbic acid (vitamin C) is a free radical scavenger that is water soluble. The daily suggested dose is 60 mg. Aside from these carotenoids, other carotenoids such as beta-carotene, lycopene, lutein, and others function as important antioxidants, quenching 1O2 and ROO•. Flavonoids, which are typically present in plants as colouring pigments, can act as powerful antioxidants at varying concentrations [5, 18, 19].

### **3.8 Antioxidants and human disease prevention**

Several epidemiological studies have discovered an inverse association between established antioxidants/phytonutrient levels in tissue/blood samples and the occurrence of cardiovascular disease, cancer, or mortality from these diseases. A recent meta-analysis, however, reveals that supplementing with largely single antioxidants may not be as beneficial. A point of view that contradicts preclinical and epidemiological data on the use of antioxidant-rich foods. Based on the majority of epidemiological and casecontrol studies, recommendations for daily dietary intake of several well-known antioxidants, such as vitamin E and C, as well as others, were produced. Because of dietary variances, antioxidant requirements in India differ from those in industrialised western nations. There are also a variety of antioxidant-rich dietary supplements that have been studied for effectiveness. Many laboratories in India are researching the antioxidant impact of plant chemicals, primarily sourced from natural sources, that can protect against such damage. Carotenoids, curcumin from turmeric, flavonoids, caffeine (found in coffee, tea, and other beverages), orientin, vicenin, glabridin, glycyrrhizin, emblicanin, punigluconin, pedunculagin, 2-hydroxy-4 methoxy benzoic acid, dehydrozingerone, picroliv, withaferin, yakuchinone, gingerol, chlorogenic acid, van (a water-soluble analogue of chlorophyll) [20].

### **3.9 Newer therapeutic approaches using antioxidants**

Over the last three decades, antioxidant-based drugs/formulations for the prevention and treatment of complicated illnesses such as atherosclerosis, stroke, diabetes, Alzheimer's disease (AD), Parkinson's disease, cancer, and others have emerged. The significance of dietary antioxidants in the prevention of numerous human illnesses, including cancer, atherosclerosis, stroke, rheumatoid arthritis, neurodegeneration, and diabetes, has been substantially influenced by free radical theory. Dietary

antioxidants may offer intriguing therapeutic potential in delaying the onset of Alzheimer's disease and its associated consequences in the elderly population. There are two neuroprotective clinical studies with antioxidants available: the Deprenyl and tocopherol antioxidant treatment of Parkinson's research. India can manufacture world-class products by combining traditional knowledge and contemporary science. As a result, it has launched a fast-track effort to develop novel pharmaceuticals by expanding on established therapies and examining the country's various plant and microbial sources. This initiative is not only the world's largest undertaking of its sort in terms of scale, variety, and access to talent and resources, but it is also unique [21].

### **3.10 Ayurveda, antioxidants and therapeutics**

Ayurvedic medications are often tailored to an individual's constitution using a unique holistic approach. Ayurvedic Indian and traditional Chinese systems are living 'great traditions,' and they play major roles in the bioprospecting of novel medications from medicinal plants, which are also high in antioxiodants. According to current estimates, around 80% of people in underdeveloped nations still rely on traditional medicine, which is mostly focused on diverse kinds of plants and animals, for their main treatment. Ayurveda is one of the most ancient and still extensively practised systems in India.

### **4. Sources of antioxidants, phytonutrients and functional foods**

Natural substances, particularly those originating from food sources, contain a considerable amount of antioxidants. Some drinks, such as tea, are also high in antioxidants. A increasing amount of research shows that moderate tea drinking may protect against several types of cancer, cardiovascular disease, kidney stone development, bacterial infections, and dental cavities. Tea is notably high in catechins, the most abundant of which is epigallocatechin gallate (EGCG) [9, 15].

### **4.1 Indian medicinal plants**

Aside from food sources, Indian medicinal plants contain antioxidants, such as: (with common/ayurvedic names in brackets) *Aegle marmelos* (Bengal quince, Bel), *Allium cepa* (Onion), *Allium sativum* (Garlic, Lahsuna), *Aloe vera* (Indian aloe, Ghritkumari), Amomum subulatum (Greater cardamom, Bari elachi), *Asparagus racemosus* (Shatavari), *Azadirachta indica* (Neem, Nimba) [15].

### **4.2 Synthetic antioxidants**

Due to availability and Importance, synthetic antioxidants are commonly employed as food additives to prevent rancidification. In edible vegetable oil and cosmetics, synthetic antioxidants such as butylated hydroxyanisole, tertiary butyl hydroquinone, 2,4,5-trihydroxybutyrophenone, octyl gallate, propyl gallate, 4-hexylresorcinol and nordihydroguaiaretic acid and are utilised [22, 23]. As synthetic phenolic antioxidants, propyl gallate and butylated hydroxyanisole shown greater chemical activity in reducing chain start of unsaturated fatty acid oxidation. Although antioxidants are effective in protecting product quality during food distribution, excessive amounts of antioxidants added to food may generate toxicities or

### *Antioxidants Obtained from the Natural Sources: Importance in Human Health DOI: http://dx.doi.org/10.5772/intechopen.109440*

mutagenicities, putting people's health at risk. The antioxidant will be chosen based on the kind of fat and oil in the diet. Butylated hydroxyanisole and butylated hydroxytoluene dissolve in most fats and oils, however they work best in animal fats. When consumed in conjunction with other meals, it has a more favourable impact than when used alone. Propyl gallate, on the other hand, which is not easily soluble, is more effective in vegetable oils than butylated hydroxyanisole and butylated hydroxytoluene, tertiary butyl hydroquinone is the most efficient antioxidant for slowing oxidation in unsaturated fats such as vegetable oils. Lower quantities of tertiary butyl hydroquinone can achieve oxidative stability than other synthetic antioxidants [24].

### **4.3 Natural and synthetic antioxidants**

Natural and synthetic antioxidants are utilised as food additives in the food business to help extend the shelf life and appearance of various foods. Synthetic phenolic antioxidants (butylated hydroxyanisole, propyl gallate and butylated hydroxytoluene [BHT]) substantially suppress oxidation; for example, chelating compounds like Metals can be bound by ethylene diamine tetraacetic acid (EDTA), reducing their contribution to the process. Antioxidants are also naturally contained in many foods and are essential for human health. They contain vitamins C and E, which may be found in fruits and vegetables and seeds and nuts, respectively. Antioxidants can be found in vitamins (ascorbic acid and -tocopherol), herbs and spices (rosemary, thyme, oregano, sage, basil, pepper, clove, cinnamon, and nutmeg), and plant extracts (tea and grapeseed). While synthetic antioxidants (such as butylated hydroxytoluene and butylated hydroxyanisole) are commonly used to protect the quality of ready-toeat food items, public concern about their safety has prompted the food industry to explore natural antioxidants. Some people's health issues have been induced by synthetic antioxidants. Butylated hydroxyanisole, butylated hydroxytoluene, and tertiary butyl hydroquinone appear to be the most troublesome antioxidants, with gallates coming in second position and having been utilised in food items with certain limits since the late 1950s. TBHQ is a relatively recent addition to the list of antioxidants permitted in food; it was approved for use as an antioxidant in food in Europe in 2004. Butylated hydroxyanisole, butylated hydroxytoluene, and tertiary butyl hydroquinone are typically found in meals containing oil and fat. Their activity is comparable to that of Vitamin E, which is utilised as an alternative antioxidant in some of the same products. These antioxidants may exist alone in a diet, but they are frequently combined with other molecules that have antioxidant action, such as phosphoric acid, propyl gallate, citric acid, and ascorbic acid [17, 25].

### **4.4 Health concerns of synthetic antioxidants**

While the bulk of studies have been conducted on animals, there is still a substantial body of research that has discovered issues with synthetic antioxidants in humans [26–28]. The **Table 1** below covers some of the human health concerns associated with butylated hydroxyanisole, butylated hydroxytoluene, and tertiary butyl hydroquinone. In one study, seven people reported symptoms such as vasomotor rhinitis, headache, flushing, asthma, conjunctival suffusion, dull retrosternal (behind the breastbone) pain radiating to the back, diaphoresis (excessive sweating), or somnolence after being exposed to butylated hydroxyanisole and butylated hydroxytoluene (sleepiness). In a subsequent trial looking for cross-reactivity with aspirin, they


### **Table 1.**

*Effect of butylated hydroxy anisole, butylated hydroxytoluene, and tertiary butyl hydroquinone on human health.*

discovered twenty-one patients who were intolerant to butylated hydroxyanisole and butylated hydroxytoluene. A handful of persons have developed dermatitis after being exposed to these synthetic antioxidants [29]. In one investigation, tertiary butyl hydroquinone in a hair colour produced contact dermatitis, and cross sensitization with butylated hydroxyanisole and butylated hydroxytoluene was seen. According to the US Department of Health and Human Services' Carcinogens report, butylated hydroxyanisole is "reasonably expected to be a human carcinogen based on substantial evidence of carcinogenicity in experimental animals." There is also worry that "butylated hydroxytoluene. May change to other carcinogenic chemicals in the human body." One conversion product of butylated hydroxytoluene (the hydroperoxide form, for example) has been demonstrated to disrupt chemical signals conveyed from cell to cell.

### **5. Health issues related to the antioxidant**

### **5.1 Neurodegenerative disorders**

Because of the high amount of lipids, particularly polyunsaturated fatty acids, nervous tissue, including the brain, is very sensitive to free radical damage. Biochemical and histological investigations in Alzheimer's disease have revealed elevated levels of oxidative stress and membrane damage. Peroxidation of Lipids Changes in antioxidant enzyme levels in neurons of Alzheimer's disease patients, such as catalase and CuZn- and Mn-SOD, are associated with increasing stress. Protein oxidation, protein nitration, and lipid peroxidation have all increased. They are seen in neurofibrillary tangles and neuritic plaques. Increased levels of peroxidation products such as 4 hydroxynonenal (4-HNE) in the cerebral fluid of Alzheimer's disease patients suggest widespread lipid peroxidation. Iron (Fe2+) is thought to play a role in enhanced lipid peroxidation in Alzheimer's disease. Multiple pathways, including impairment of the activity of membrane ion-motive ATPases (Na+/K + -ATPase and Ca2 + -ATPase), glucose transporters, and glutamate transporters, may contribute to neuronal mortality in Alzheimer's disease. Lipid peroxidation produces the aldehyde 4-HNE, which appears to play a key role in the neurotoxic effects of amyloid [14].

### **5.2 Free radicals, diabetes and ages**

Experimental data suggests that free radicals have a role in the establishment of diabetes and, more crucially, the development of diabetic complications [30]. The

### *Antioxidants Obtained from the Natural Sources: Importance in Human Health DOI: http://dx.doi.org/10.5772/intechopen.109440*

Free radical scavengers are useful in avoiding experimental diabetes in animal models and in type 1 (IDDM) and type 2 (NIDDM) patients, as well as in lowering the severity of diabetic sequelae. Persistent hyperglycemia in diabetic individuals causes oxidative stress due to


The spontaneous reduction of molecular oxygen to superoxide and hydroxyl radicals, which are extremely reactive and interact with all biomolecules, occurs during glucose auto-oxidation. In addition, they hasten the production of advanced glycation end products (AGEs). Pyrroles and imidazoles, for example, tend to accumulate in tissue. Crosslinking AGE-protein with other macromolecules in tissues causes cell and tissue function problems. The third route by which free radicals are created in tissues is the polyol pathway [31]. This route deletes 30 lots of NADPH, which reduces the production of antioxidants like glutathione. The ability of antioxidant enzymes is also diminished as a result of protein glycation. In endothelial cells, free radicals react with nitric oxide, resulting in a reduction of vasodilation function. Long-lived structural proteins, collagen and elastin, undergo non-enzymatic crosslinking throughout life and in diabetics [32]. This abnormal protein crosslinking is mediated by AGEs generated by nonenzymatic glycosylation of proteins by glucose.

### **5.3 Free radical damage to DNA and cancer**

DNA is a common site of free radical damage. Strand breakage (single or double strand breaks), different forms of base damage giving products such as 8 hydroxyguanosine, thymine glycol, or abasic sites, damage to deoxyribose sugar, and DNA protein cross linkages are among the numerous types of damages generated. These damages can cause heritable changes in the DNA, which can lead to cancer in somatic cells or foetal abnormalities in germ cells. The interaction of free radicals with tumour suppressor genes and proto-oncogenes suggests that they have a role in the genesis of several human malignancies [9]. Cancer occurs as a result of a series of genetic alterations. Tobacco smoking and chewing, UV rays from sunshine, radiation, viruses, chemical contaminants, and other factors can all be initiating agents. Hormones are examples of promoting agents (androgens for prostate cancer, estrogens for breast cancer and ovarian cancer). Inflammation causes the production of iNOS (inducible nitric oxide synthase), as well as COX and LOX. These are capable of initiating carcinogenesis. Experimental and epidemiological evidence show that a number of dietary components can serve as antioxidants, inhibiting cancer formation and lowering cancer risk. Vitamins A, C, E, beta-carotene, and micronutrients such as antioxidants and anticarcinogens are among them [33, 34]. The recent studied the processes behind dietary phytochemical anticancer effects. Chemopreventive phytochemicals have the ability to prevent or reverse the promotion stage of multistep carcinogenesis. They can also prevent or slow the growth of precancerous cells into malignant cells. Many of the molecular changes linked with carcinogenesis occur in cell-signalling pathways that control cell proliferation and differentiation. The family of mitogen activated protein kinases is a key component of the intracellular signalling network that maintains homeostasis (MAPKs). With the activation of the transcription factors NF-B and AP1, several intracellular signal-transduction pathways converge. These variables are prominent targets of several types of chemopreventive phytochemicals because they mediate the pleiotropic effects of both external and internal stimuli in the cellular-signalling cascades [34]. Curcumin, the active ingredient of *Curcuma longa* (Turmeric, Haldi), inhibits the production of COX2, LOX, iNOS, MMP-9, TNF, chemokines, and other cell-surface adhesion molecules, as well as cyclin D1. Human clinical studies have demonstrated that curcumin at dosages up to 10 g/day is safe and can inhibit tumour start, promotion, and metastasis. Many Long-term prospective clinical investigations are required to validate the hypothesis.

### **5.4 Mitochondria, oxidative protein damage and proteomics**

Proteomic technologies' fast advancement and application to large-scale investigations of protein-protein interactions and protein expression patterns imply that these approaches are ideally suited to give the molecular insights required to completely comprehend oxidative harm caused by free radicals [35]. The very significant progress has been made in identifying specific proteins that are confined to the mitochondria throughout the previous two decades. Specifically, the 100 or more subunits that make up the five complexes of the electron transport chain (ETC). Several groups have recently begun to tackle the bigger task of establishing the content of complete mitochondrial proteomes from a variety of key model systems as well as human tissues, utilising contemporary mass spectrometry (MS)-based proteomic methods. Gibson and colleagues identified 684 distinct proteins from the combined peptide data acquired from over 100,000 mass spectra generated by MALDI-MS and high performance liquid chromatography (HPLC) MS/MS analysis using mitochondria isolated from human heart. These findings have been included into 'MitoProteome,' a publicly available database for the human heart mitochondrial proteome.

### **5.5 Free radicals and ageing**

Ageing is caused by mitochondrial ROS generation and oxidative damage to mitochondrial DNA. Increased lipid peroxidation in cellular membranes as a result of oxidative stress results in fatty acid unsaturation. According to the most current study on 'free radicals and ageing, Caloric restriction (CR) is the only known experimental alteration that reduces the rate of mammalian ageing, and it has multiple beneficial impacts on rodent and likely human brains. Calorie-restricted mitochondria, like those seen in long-lived animal species, effectively inhibit ROS generation with pyruvate and malate at complex I. The oxygen consumption of the mitochondria stays same, while the free radical leak from the electron transport chain is reduced in CR. Many researchers discovered that increasing the number of oxidative stress defence systems may increase an organism's health span. Arking's group's work on artificial selection in flies resulted in organisms with much higher levels of oxidative stress tolerance and more efficient mitochondria. Indeed, lower ROS formation and increased ROS removal resulted in less oxidative damage and a later start of senescence in those flies. However, using genetic engineering techniques to introduce additional copies of these oxidative stress-resistance genes into mice did not result in a longer lifetime.

*Antioxidants Obtained from the Natural Sources: Importance in Human Health DOI: http://dx.doi.org/10.5772/intechopen.109440*

### **6. Conclusion**

Natural ingredients are becoming increasingly popular these days. Food antioxidants and preservatives may cause lipid peroxidation and deterioration of taste and quality. Free radicals have been linked to the genesis of a wide range of important illnesses. They can cause several important biological molecules to lose shape and function. Such unfavourable alterations in the body might result in illness. Antioxidants can guard against the damage caused by free radicals at various levels of action. Plants' dietary and other components are rich in antioxidants. The link between free radicals, antioxidants, and the function of numerous organs and organ systems is extremely complicated, and the discovery of 'redox signalling' marks a watershed moment in this critical interaction. Recent research has focused on several ways for protecting vital tissues and organs from oxidative damage caused by free radicals. Many unique techniques have been developed, and substantial discoveries have been achieved in recent years. Natural antioxidants are abundant in the traditional Indian food, spices, and medicinal herbs. Increased consumption of foods with functional properties, such as high levels of antioxidants in functional foods, is one method that is gaining traction in advanced nations and is making an appearance in our country.

This chapter focuses on an overview of the potentials of numerous sources with appropriate antioxidant potential, as well as their influence on human health. Because 70–80% of the world's population cannot afford contemporary supplements and treatments, this chapter illustrates that individuals may priorities their food habits depending on the antioxidant capacity and cost-effectiveness of the accessible supply.

### **Conflict of interest**

None.

### **Author details**

Sushil S. Burle, Krishna R. Gupta\*, Swati N. Lade, Shyam W. Rangari and Milind J. Umekar Smt. Kishoritai Bhoyar College of Pharmacy, New Kamptee, Nagpur, India

\*Address all correspondence to: krg1903@gmail.com

© 2023 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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Section 2
