Diallyl sulfide, a sulfur-containing volatile compound present in garlic (Allium sativum), exerts anticarcinogenic activity in various rodent tumor models. In the present study, apoptosis-inhibiting effects of diallyl sulfide against a carcinogenic polycyclic aromatic hydrocarbon, 7,12-dimethyl benz(a)anthracene (DMBA), in Swiss albino mice were observed. The animals were given either 250 μg/mouse or 500 μg/mouse of diallyl sulfide for 1 week after a single intragastric dose of 7,12-dimethyl benz(a)anthracene (50 mg/kg body weight). Results showed that diallyl sulfide supplementation effectively protects against 7,12-dimethyl benz(a)anthracene—induced oxidative stress, characterized by restored antioxidant enzyme levels (up to 64%) and lipid peroxidation (up to 25%). Flow cytometric analysis showed a reduction in apoptotic cell population in hypodiploid region in diallyl sulfide–supplemented animals. Inhibition of apoptosis was preceded by decrease in reactive oxygen species levels and restoration of mitochondrial transmembrane potential followed by decreased DNA fragmentation. In 7,12-dimethyl benz(a)anthracene–exposed animals, downregulation (~30%) of antiapoptotic Bcl-2 and upregulation (~60%) of pro-apoptotic Bax proteins were observed. These alterations were restored significantly by diallyl sulfide supplementation, indicating inhibition of apoptosis. Thus, these results show that diallyl sulfide provides protection against oxidative damage induced by 7,12-dimethyl benz(a)anthracene in mouse liver and may be an effective chemopreventive and therapeutic agent by modulating expression of cell-growth regulatory proteins.
1 Thilly WG . Have environmental mutagens caused oncomutations in people?Nat Genet2003; 34: 255 – 259 .
2.
2 Duthie SJ . Berry phytochemicals, genomic stability and cancer: evidence for chemoprotection at several stages in the carcinogenic process.Mol Nutr Food Res2007; 51: 665 – 674 .
3.
3 Abdulla MGruber P . Role of diet modification in cancer prevention.Biofactors2000; 12: 45 – 51 .
4.
4 Stoner GDChen TKresty LAAziz RMReinemann TNines R . Protection against esophageal cancer in rodents with lyophilized berries: potential mechanisms.Nutr Cancer2006; 54: 33 – 46 .
5.
5 Barth SWFahndrich CBub ADietrich HWatzl BWill F . Cloudy apple juice decreases DNA damage, hyperproliferation and aberrant crypt foci development in the distal colon of DMH-initiated rats.Carcinogenesis2005; 26: 1414 – 1421 .
6.
6 Middleton JKandaswami CTheoharides T . Effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer , Pharmaco Res2000; 52: 673 – 751 .
7.
7 Ferguson L . Role of plant polyphenols in genomic stability.Mutat Res2001; 475: 89 – 111 .
8.
8 Merillon JFauconneau BHuguel F . Antioxidant activity of the stilbene astringing, newly extracted from Vitis vinifera cell culture.Clin. Chem1997; 43: 1092 – 1093 .
9.
9 Bentivegna SWhitney K . Subchronic 3-month oral toxicity study of grape seed and grape skin extracts , Food Chem Toxicol2002; 40: 1731 – 1743 .
10.
10 Wargovich MJImada OStephens LC . Initiation and post initiation chemopreventive effects of diallyl sulfide in esophageal carcinogenesis.Cancer Lett.1992; 64: 39 – 42 .
11.
11 Prasad SKalra NShukla Y . Modulatory effects of diallyl sulfide against testosterone-induced oxidative stress in Swiss albino mice.Asian J Androl2006; 8: 719 – 723 .
12.
12 Song KMilner JA . Heating garlic inhibits its ability to suppress 7,12 dimethylbenz(a)anthracene-induced DNA adduct formation in rat mammary tissue.J Nutr1999; 129: 657 – 661 .
13.
13 Lin JGChen GWSu CCHung CFYang CCLee JH . Effects of garlic components diallyl sulfide and diallyl disulfide on arylamine N-acetyltransferase activity and 2-aminofluorene-DNA adducts in human promyelocytic leukemia cells.Am J Chin Med2002; 30: 315 – 325 .
14.
14 Kalra NArora AShukla Y . Involvement of multiple signaling pathways in diallyl sulfide mediated apoptosis in mouse skin tumors.Asian Pac J Cancer Prev2006; 7: 556 – 562 .
15.
15 Arora ASiddiqui IAShukla Y . Modulation of p53 in 7,12-dimethylbenz[a]anthracene-induced skin tumors by diallyl sulfide in Swiss albino mice.Mol Cancer Ther2004; 3: 1459 – 1466 .
16.
16 Singh AArora AShukla Y . Modulation of altered hepatic foci induction by diallyl sulphide in Wistar rats.Eur J Cancer Prev2004; 13: 263 – 269 .
17.
17 Wu CCChung JGTsai SJYang JHSheen LY . Differential effects of allyl sulfides from garlic essential oil on cell cycle regulation in human liver tumor cells.Food Chem Toxicol2004; 42: 1937 – 1947 .
18.
18 Flora SJ . Role of free radicals and antioxidants in health and disease.Cell Mol Biol (Noisy-le-grand)2007; 53: 1 – 2 .
19.
19 Bickers DRAthar M . Oxidative stress in the pathogenesis of skin disease.J Invest Dermatol2006; 126: 2565 – 2575 .
20.
20 Richter CPark JWAmes BN . Normal oxidative damage to mitochondrial and nuclear DNA is extensive.Proc Natl Acad Sci USA. 1988; 85: 6465 – 6467 .
21.
21 Sauer HWartenberg MHescheler J . Reactive oxygen species as intracellular messengers during cell growth and differentiation.Cell Physiol Biochem2001; 11: 173 – 186 .
22.
22 Stewart JHTran TLLevi NTsai WSSchrump DSNguyen DM . The essential role of the mitochondria and reactive oxygen species in cisplatin-mediated enhancement of fas ligand-induced apoptosis in malignant pleural mesothelioma.J Surg Res2007; 141: 120 – 131 .
23.
23 Reed JC . Bcl-2 and the regulation of programmed cell death.J Cell Biol1994; 124: 1 – 6 .
24.
24 Takagi-Morishita YYamada NSugihara AIwasaki TTsujimura TTerada N . Mouse uterine epithelial apoptosis is associated with expression of mitochondrial voltage-dependent anion channels, release of cytochrome C from mitochondria, and the ratio of Bax to Bcl-2 or Bcl-X.Biol Reprod2003; 68: 1178 – 1184 .
26 Sinha AK . Colorimetric assay of catalase.Anal Biochem1972; 47: 389 – 394 .
27.
27 Carlberg IMennervick B . Glutathione reductase. In: Methods in enzymology.New York : Acadmic Press ; 1985. p. 484–490.
28.
28 Habig WHPabst MJJakoby WB . Glutathione-S-transferases, the first enzymatic step in mercapturic acid formation.J Biol Chem1974; 249: 130 – 139 .
29.
29 Ohkawa HOhisi NYagi K . Assay for lipid peroxides in animal tissues by thiobarbutric acid reaction.Anal Biochem1979; 95: 351 – 358 .
30.
30 Lowry OHRosebrough NJFarr ALRandall RJ . Protein measurement with Folin phenol reagent.J Biol Chem1951; 193: 265 – 275 .
31.
31 Esposti DMMcLennan H . Mitochondria and cells produce reactive oxygen species in virtual anaerobiosis: relevance to ceramide-induced apaotosis.FEBS Lett1998; 430: 338 – 342 .
32.
32 Prasad SKalra NShukla Y . Hepatoprotective effects of lupeol and mango pulp extract in carcinogen induced alteration in Swiss albino mice.Mol Nutr Food Res2007; 51: 352 – 359 .
33.
33 Nicoletti IMigliorati GPagliacci MCGrignani FRiccardi C . A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry.J Immunol Methods1991; 139: 271 – 279 .
34.
34 Bai JRodriguez AMMelendez JACederbaum AI . Overexpression of catalase in cytosolic or mitochondrial compartment protects HepG2 cells against oxidative injury.J Biol Chem1999; 274: 26217 – 26224 .
35.
35 Arora ASiddiqui IAShukla Y . Modulation of p-glycoprotein mediated multidrug resistance in K562 leukemic cells by indole-3-carbinol.Toxicol Appl Pharmacol2005; 202: 237 – 243 .
36.
36 Hastak KAgarwal MKMukhtar HAgarwal ML . Ablation of either p21 or Bax prevents p53-dependent apoptosis induced by green tea polyphenol epigallocatechin-3-gallate.FASEB J2005; 19: 789 – 791 .
37.
37 Devasagyam TPKamat JP . Biological significance of singlet oxygen.Indian J Exp Biol2002; 40: 680 – 692 .
38.
38 Behrend LHenderson GZwacka RM . Reactive oxygen species in oncogenic transformation.Biochem Soc Trans2003; 31: 1441 – 1444 .
39.
39 Melov S . Therapeutics against mitochondrial oxidative stress in animal models of aging.Ann NY Acad Sci2002; 959: 330 – 340 .
40.
40 Maire MARast CPagnout CVasseur P . Changes in expression of bcl-2 and bax in Syrian hamster embryo (SHE) cells exposed to ZnCl2.Arch Toxicol2005; 79: 90 – 101 .
41.
41 Yoshida YIzumi HTorigoe TIshiguchi HItoh HKang D . P53 physically interacts with mitochondrial transcription factor A and differentially regulates binding to damaged DNA.Cancer Res2003; 63: 3729 – 3734 .
42.
42 Vang ORasmussen BFAndersen O . Combined effects of complex mixtures of potentially anti-carcinogenic compounds on antioxidant enzymes and carcinogen metabolizing enzymes in the rat.Cancer Lett1997; 114: 283 – 286 .
43.
43 Weisburger JH . Antimutagenesis and anticarcinogenesis from the past to the future.Mutat Res2001; 480: 23 – 35 .
44.
44 Pigeolet ECorbisier PHoubion ALambert DMichiels CRaes M . Glutathione peroxidase, superoxide dismutase, and catalase inactivation by peroxides and oxygen derived free radicals.Mech Ageing Dev1990; 51: 283 – 297 .
45.
45 Kalra NPrasad SShukla Y . Antioxidant potential of black tea against 7,12-dimethylbenz(a)anthracene-induced oxidative stress in Swiss albino mice.J Environ Pathol Toxicol Oncol2005; 24: 105 – 114 .
46.
46 Locigno RCastronovo V . Reduced glutathione system: role in cancer development, prevention and treatment.Int J Oncol2001; 19: 221 – 236 .
47.
47 Chung FLChen HJNath RG . Lipid peroxidation as a potential endogenous source for the formation of exocyclic DNA adducts.Carcinogenesis1996; 10: 2105 – 2111 .
48.
48 Valko MRhodes CJMoncol JIzakovic MMazur M . Free radicals, metals and antioxidants in oxidative stress-induced cancer.Chem Biol Interact2006; 160: 1 – 40 .
49.
49 Storz GPolla BS . Transcriptional regulators of oxidative stress inducible genes in prokaryotes and eukaryotes.Exp Suppl Basel, 1996; 77: 239 – 254 .
50.
50 Maldonado PDBarrera DRivero IMata RMedina-Campos ONHernandez-Pando R . J. Antioxidant S-allylcysteine prevents gentamicin-induced oxidative stress and renal damage.Free Radic Biol Med2003; 35: 317 – 324 .
