Basic nutritional investigationEffect of garlic on lipid peroxidation and antioxidation enzymes in DMBA-induced skin carcinoma
Introduction
Over the centuries, garlic has acquired a special position in the folklore of many cultures as a formidable prophylactic and therapeutic medicinal agent. Recent research on dietary factors has demonstrated that they play a key role in the development of various human cancers, including skin carcinogenesis. Several epidemiologic studies have indicated that certain diets are associated with a low risk of human diseases and that these diets are rich in fruits, herbs, and spices; the common spice among them is garlic [1]. Considerable anecdotal evidence has supported the invaluable role that garlic has in the therapy of many diseases [2], [3], [4], [5], [6], [7]. Garlic has attracted particular attention by modern medicine because of its widespread use around the world and the cherished belief that it helps in maintaining good health by warding off illnesses and providing more vigor. Papers presented at the 2005 Garlic Symposium in Washington, D.C., held April 9 to 11, reported the current scientific research into the effects of garlic on heart health and cancer prevention. These biological responses from garlic have been largely attributed to 1) reduction of risk factors for cardiovascular diseases and cancer, 2) stimulation of immune function, 3) enhanced detoxification of foreign compounds, 4) hepatoprotection, 5) antimicrobial effects, and 6) antioxidant effects. We report on the beneficial effect of garlic on skin carcinogenesis. Skin cancers are the fastest growing type of cancer in the United States, surpassing lung, breast, colorectal, and prostate cancers. More than 1 million Americans were diagnosed with skin cancer in 2007 [8]. The scavenger functions of the cells through phase II enzymes and other antioxidants are required for proper homeostasis of the skin; otherwise it will develop into the deadliest form of skin cancer—the malignant melanoma. This form of skin cancer causes the major part of all skin cancer fatalities.
An aqueous solution of fresh garlic contains many substances that boost several antioxidant and phase II enzymes in the body and act in concert to prevent disease and age-related conditions. Garlic contains water-soluble allyl amino acid derivatives, which account for most of its organosulfur content, lipid-soluble allyl sulfides (allicin), flavonoids, saponins, and essential macro- and micronutrients that demonstrate antioxidant activity [9]. Allicin is believed to be the main component responsible for the biological activity of garlic [10]. Other lipid-soluble compounds in garlic include diallyl sulfide [11], triallyl sulfide, diallyl disulfide, diallyl polysulfides, and others [12], [13]. Other antioxidants in garlic include phenolic compounds, notably alexin, whose phenolic hydroxyl group confers antioxidant activity, N-fructosyl glutamate, N-fructosyl arginine, and selenium [14].
Oxidative stress (free radical overload and insufficient antioxidants) and inflammation play a role in many cancers. Accumulation of oxidative stress–related damaged proteins and their improper removal is a hallmark of many cancers including skin cancer [15], [16], [17]. Reactive oxygen species (ROS) are produced during several intracellular pathways. ROS are increased during infection, inflammation, and exposure to pollutants, radiation, and sunlight. These oxygen radicals eventually cause damage to cellular DNA and protein and subsequently foster tumor growth in many types of cancer [17], [18], [19], [20]. Several phytochemicals have been demonstrated to decrease lipid peroxides (LPOs) and maintain the redox status of intracellular enzymes that are needed for homeostasis of the cells [21], [22]. Preclinical studies have shown that garlic prevents many types of cancer by disabling free radicals, blocking carcinogen–DNA binding, and increasing carcinogen disposal [14], [23], [24].
Garlic extract also boosts some immune functions and selectively kills cancer cells by apoptosis [25], [26]. Garlic extract inhibits early and late stages of carcinogenesis, resulting in inhibition of tumor growth in many tissues, including the colon, mammary glands, skin, stomach, and esophagus [9], [12], [24], [27], [28], [29]. In this study, we found that an aqueous infusion of garlic can prevent the incidence of 7,12 dimethylbenz[a]anthracene (DMBA)-induced skin carcinogenesis in female swiss albino mice. The garlic-fed mice showed a reduction in tumor size due to reduction of LPO formation and simultaneous stimulation of phase II detoxifying enzymes and other antioxidants. This study will strengthen the importance of garlic ingestion in a regular diet to maintain a healthy body and during chemoprevention of skin carcinoma.
Section snippets
Chemicals
The DMBA, croton oil (1-chloro-2-4-dinitrobenzene), glutathione (GSH), glutathione reductase (GR), nicotinamide adenine dinucleotide phosphate reduced form (NADPH), pyrogallol, thiobarbituric acid (TBA), diethylene triamine penta acetic acid, and pyridine were purchased from Sigma Chemicals Co. (St. Louis, MO, USA). Sodium dodecylsulfate was purchased from Gibco BRL (USA), and hydrogen peroxide solution (H2O2; 30%), acetic acid (CH3COOH), n-butanol, and potassium chloride (KCl) from Merck,
Effects of aqueous infusion of garlic on body weight of DMBA-induced mice
Thirty female “Swiss albino mice” were separated into five different groups (Table 1), so that each group had six mice in total. All groups of mice were treated separately as presented in Table 1 with the NC group of mice serving as a control to the entire experimental set. All 30 mice had a starting body weight of 22 ± 1 g (Fig. 1). Tumors in the swiss albino mice were generated as it was described next to the CC group in Table 1. The NC group received only acetone treatment (Fig. 2A). A
Discussion
The transformation of normal cells to the malignant state proceeds through several discernible stages, including initiation by DNA damage and later events that have been defined as tumor promotion in animals and in vitro [23], [44]. Cancer chemoprevention by phytochemicals appears to be one of the most feasible approaches for cancer control. Vegetables, fruits, spices, teas, herbs, and medicinal plants, such as carotenoids, phenolic compounds, and terpenoids, have been proven to suppress
Conclusion
This report demonstrates the superior chemopreventive effect of garlic against skin carcinogenesis. Garlic induced elevated levels of antioxidants, which was accompanied by decreased lipid peroxidation and incidences of papillomas. Garlic treatments also downregulated COX-2 expression, a cancer progression marker, and decreased p53 and caspase-3 expression.
Acknowledgments
Dr. Das thanks the director, Dr. Jaydeep Biswas, and all the other staff members of the Department of Cancer Chemoprevention, Chittaranjan National Cancer Institute, for their support and encouragement. Dr. Saha thanks the Department of Oncology, Lombardi Comprehensive Cancer Center, for its available infrastructure and support.
References (71)
Role of chemopreventers in human diet
Clin Biochem
(1994)- et al.
The medical uses of garlic—fact and fiction
Am Pharm
(1982) - et al.
Inhibiting progression of coronary calcification using aged garlic extract in patients receiving statin therapy: a preliminary study
Prev Med
(2004) Ajoene (natural garlic compound): a new anti-leukaemia agent for AML therapy
Leuk Res
(2004)- et al.
Dietary components modify the ability of garlic to suppress 7,12-dimethylbenz(a)anthracene-induced mammary DNA adducts
J Nutr
(1996) - et al.
Allicin, a major component of garlic, inhibits apoptosis of macrophage in a depleted nutritional state
Nutrition
(2006) Oxidative stress and apoptosis: impact on cancer therapy
J Pharm Sci
(2007)- et al.
Free radicals, metals and antioxidants in oxidative stress-induced cancer
Chem Biol Interact
(2006) - et al.
Modulation of xenobiotic-metabolizing enzymes and redox status during chemoprevention of hamster buccal carcinogenesis by bovine lactoferrin
Nutrition
(2006) - et al.
Effects of dietary fish oil on lipid peroxidation and serum triacylglycerol levels in psychologically stressed mice
Nutrition
(2008)
Inhibition of cell growth and induction of apoptosis via inactivation of NF-kappaB by a sulfur compound isolated from garlic in human colon cancer cells
J Pharmacol Sci
Cancer chemoprevention with garlic and its constituents
Cancer Lett
Protein measurement with the Folin phenol reagent
J Biol Chem
Increased levels of lipid peroxides in aged rat brain as revealed by direct assay of peroxide values
Neurosci Lett
Glutathione S-transferasesThe first enzymatic step in mercapturic acid formation
J Biol Chem
Superoxide dismutaseAn enzymic function for erythrocuprein (hemocuprein)
J Biol Chem
A spectrophotometric method for determination of catalase activity in small tissue samples
Anal Biochem
Automated assays for superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase activity
Anal Biochem
Modulation of chemical carcinogenesis by xenobiotics
Fundam Appl Toxicol
Phytochemicals: guardians of our health
J Am Diet Assoc
GST genotypes and lung cancer susceptibility in Asian populations with indoor air pollution exposures: a meta-analysis
Mutat Res
Mammalian class theta GST and differential susceptibility to carcinogens: a review
Mutat Res
Occupational exposure to antineoplastic agents induces a high level of chromosome damageLack of an effect of GST polymorphisms
Toxicol Appl Pharmacol
Effects of peroxisome proliferators on glutathione and glutathione-related enzymes in rats and hamsters
Toxicol Appl Pharmacol
Catalase transgenic mice: characterization and sensitivity to oxidative stress
Arch Biochem Biophys
Aged garlic extract may be safe for patients on warfarin therapy
J Nutr
Garlic extract therapy in children with hypercholesterolemia
Arch Pediatr Adolesc Med
Aged garlic extract is a potential therapy for sickle-cell anemia
J Nutr
Common cancer types
Indian food ingredients and cancer prevention—an experimental evaluation of anticarcinogenic effects of garlic in rat colon
Asian Pac J Cancer Prev
Impact of various sources of garlic and their constituents on 7,12-dimethylbenz[a]anthracene binding to mammary cell DNA
Carcinogenesis
Identified diallyl polysulfides from an aged garlic extract which protects the membranes from lipid peroxidation
Planta Med
Garlic compounds protect vascular endothelial cells from oxidized low density lipoprotein-induced injury
J Pharm Pharmacol
Role of oxidative stress from mitochondria on aging and cancer
Cornea
Oxidative stress and wasting in cancer
Curr Opin Clin Nutr Metab Care
Cited by (44)
Usnic acid induced changes in biomolecules and their association with apoptosis in squamous carcinoma (A-431) cells: A flow cytometry, FTIR and DLS spectroscopic study
2022, Spectrochimica Acta - Part A: Molecular and Biomolecular SpectroscopyCitation Excerpt :Significant changes in cell surface lipid and protein molecules were observed in ursolic acid treated cholangiocarcinoma cells leading to apoptosis [11]. Earlier, phytochemicals obtained from garlic played an inhibitory role in skin carcinoma of mice by inhibiting the oxidative modification of lipid molecules, thereby protecting the healthy cells from oxidative damage [12]. Usnic acid (UA) [(R)-1,1′-(1,7,9-trihydroxy-8,9b-dimethyl-3-oxo-3,9b-dihydrodibenzo[b,d]furan-2,6-diyl)diethanone; C18H16O7, (Fig. 1) is a naturally occurring secondary metabolite, found in many lichens as a yellow pigment.
Garlic constituents for cancer prevention and therapy: From phytochemistry to novel formulations
2022, Pharmacological ResearchCitation Excerpt :Garlic restores the p53 by activating the p53 pathway in A375 and BCC cancer cells. DAS exerted its antitumorigenic effects through several mechanisms of action, such as activating carcinogen metabolism, suppressing carcinogen-induced damage to DNA, promoting the cell defense mechanism, and inducing apoptosis in carcinogen-induced skin tumors [200–202,206–210]. Alginate-based black garlic nanoparticles showed cytotoxicity against skin cancer SK-Mel 2 (HTB – 68™) cell line, where it exhibited 54.3 ± 8.4% inhibition, but the concentration was not reported [153].
Anticancer potential of garlic and its bioactive constituents: A systematic and comprehensive review
2021, Seminars in Cancer BiologyAlliin interacts with DMPC model membranes to modify the membrane dynamics: FTIR and DSC Studies
2017, Vibrational SpectroscopyCitation Excerpt :This process affects membrane properties such as membrane fluidity and cellular functions. Hence, alliin-induced fluidizing effect may be important to shift the effect of lipid peroxidation, which accounts for a reduction in lipid dynamics [44–47]. In summary, the results of the effect of alliin on the phase transition profile and the structural and dynamic properties of DMPC liposomes explain the molecular interactions of alliin with membrane systems.
Protective effect of allyl methyl disulfide on acetaminophen-induced hepatotoxicity in mice
2016, Chemico-Biological InteractionsCitation Excerpt :Quantification of lipid peroxidation is essential to assess oxidative stress in pathophysiological processes. HNE is the oxidation product of polyunsaturated fatty acid and is a major cytotoxic end product of lipid peroxidation that mediates oxidative stress-induced death in many cell types [43,44]. Results of immunohistochemical analysis reveal that the expression of HNE was highly increased in the APAP-exposed mice, and this elevation was significantly attenuated by AMDS (Fig. 5B).