Investigational New Drugs

, Volume 28, Issue 3, pp 251–259 | Cite as

Chemopreventive efficacy of gallic acid, an antioxidant and anticarcinogenic polyphenol, against 1,2-dimethyl hydrazine induced rat colon carcinogenesis

  • Jebakkan Senapathy Giftson
  • Sathiavelu Jayanthi
  • Namasivayam Nalini


Colon cancer is a major cause of morbidity and mortality in developed and developing countries and its etiology is known to be a combination of hereditary, environmental, dietary factors and lack of physical activity. Chemoprevention offers a novel approach to control the incidence of colon cancer. Gallic acid (GA) is a polyphenol widely present in tea and other plants which is popularly used in the traditional medicine of China. The present study was to evaluate the efficacy of GA supplementation on tissue lipid peroxidation and antioxidant defense system in 1,2-dimethyhydrazine (DMH) induced colon carcinogenesis in male Wistar rats. The rats were assorted into six groups, viz., group1 control rats received modified pellet diet; group 2 rats received GA (50 mg/kg body weight) orally along with modified pellet diet; group 3 rats received DMH (20 mg/kg body weight) subcutaneously once a week for the first 15 weeks; groups 4, 5 and 6 rats received GA along with DMH during the initiation, post- initiation stages and the entire period of study respectively. All the rats were sacrificed at the end of 30 weeks and the tissues were evaluated biochemically. We observed decreased lipid peroxidation (LPO) products such as thiobarbituric acid reactive substances (TBARS), lipid hydroperoxides (LOOH) and conjugated dienes (CD) and diminished levels of antioxidants such as superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione reductase (GR) and glutathione peroxidase (GPx) in the tissues of DMH treated rats, which were elevated significantly on GA supplementation. Moreover, enhanced activity of ascorbic acid and α-tocopherol levels were also observed in DMH alone treated rats which were significantly reduced on GA supplementation. Our results suggest that GA could exert a significant chemopreventive effect on DMH induced colon carcinogenesis.


Colon cancer 1,2-Dimethyl hydrazine Gallic acid Antioxidants Lipid peroxidation 


  1. 1.
    Shike M, Winamer SJ, Greenwald PH, Bolch A, Hill M, Swaroop SV (1998) Primary prevention of colorectal cancer. Bull World Health Org 68:377–385Google Scholar
  2. 2.
    Greenlee RT, Murray T, Bolden S, Wingo PA (2000) Cancer statistics 2000. CA Cancer J Clin 50:7–33CrossRefPubMedGoogle Scholar
  3. 3.
    Yamamoto S (2000) Cancer statistics digest. All cancer mortality of prefectures in Japan. Jpn J Clin Oncol 30:168PubMedGoogle Scholar
  4. 4.
    Goel A, Arnold CN, Boland CR (2001) Multistep progression of colorectal cancer in the setting of microsatellite instability: new details and novel insights. Gastroenterology 121:1497–1502CrossRefPubMedGoogle Scholar
  5. 5.
    Goel A, Nagasaka T, Arnold CN, Inoue T, Hamilton C, Niedzwiecki D, Compton C, Mayer RJ, Goldberg R, Bertagnolli MM, Boland CR (2007) The CpG island methylator phenotype and chromosomal instability are inversely correlated in sporadic colorectal cancer. Gastroenterology 132:127–138CrossRefPubMedGoogle Scholar
  6. 6.
    Bartsch H, Nair J (2002) Potential role of lipid peroxidation derived DNA damage in human colon carcinogenesis: studies on exocyclic base adducts as stable oxidative stress markers. Cancer Detect Prev 26:308–312CrossRefPubMedGoogle Scholar
  7. 7.
    Boland CR, Luciani MG, Gasche C, Goel A (2005) Infection, inflammation, and gastrointestinal cancer. Gut 54:1321–1331CrossRefPubMedGoogle Scholar
  8. 8.
    Breimer LH (1990) Molecular mechanisms of oxygen radical carcinogenesis and mutagenesis: the role of DNA base damage. Mol Carcinogen 3:188–197CrossRefGoogle Scholar
  9. 9.
    Davies KJ (1995) Oxidative stress: the paradox of aerobic life. Biochem Soc Symp 6:1–31Google Scholar
  10. 10.
    Tada M (2000) Biological activities of antioxidants from herbs in Labiatae. Foods Food Ingred J Jpn 184:33–39Google Scholar
  11. 11.
    Fiala ES (1977) Investigations into the metabolism and mode action of the colon carcinogen 1,2- dimethylhydrazine and azoxymethane. Cancer 40:2436–2445CrossRefPubMedGoogle Scholar
  12. 12.
    Fiala ES, Sohn OS, Hamilton SR (1987) Effects of chronic dietary ethanol on the in vivo and in vitro metabolism of methylazoxymethanol and methylazoxymethanol induced DNA methylation in the rat colon and liver. Cancer Res 47:5939–5943PubMedGoogle Scholar
  13. 13.
    Choudhary G, Hansen H (1998) Human health perspective on environmental exposure to hydrazines: a review. Chemosphere 37:801–843CrossRefPubMedGoogle Scholar
  14. 14.
    Sun Y (1990) Free radicals, antioxidant enzymes and carcinogenesis. Free Radic Biol Med 8:583–599CrossRefPubMedGoogle Scholar
  15. 15.
    Gower J (1988) A role for dietary lipids and antioxidants in the activation of carcinogens. Free Radic Biol Med 5:95–111CrossRefPubMedGoogle Scholar
  16. 16.
    Blakey DH, Duncan AM, Wargovich MJ, Goldberg MT, Bruce WR, Heddle JA (1985) Detection of nuclear anomalies in the colonic epithelium of the mouse. Cancer Res 45:242–249PubMedGoogle Scholar
  17. 17.
    Ma QY, Williamson KE, Rowlands BJ (2002) Variability of cell proliferation in the proximal and distal colon of normal rats and rats with dimethylhydrazine induced carcinogenesis. World J Gastroenterol 8:847–852PubMedGoogle Scholar
  18. 18.
    Halline AG, Dudeja PK, Lashner BA, Brasitas TA (1989) Urinary excretion of NV-acetylspermidine and other acetylated and free polyamines in 1,2-dimethylhydrazine model of experimental rat colon cancer. Cancer Res 49:4721–4723PubMedGoogle Scholar
  19. 19.
    Steinmetz KA, Potter JD (1991) Vegetables, fruits and cancer, I. Epidemiology. Cancer Causes Control 2:325–357CrossRefPubMedGoogle Scholar
  20. 20.
    Kahkonen MP, Hopia AI, Vuorela HJ et al (1999) Antioxidant activity of plant extracts containing phenolic compounds. J Agric Food Chem 47:3954–3962CrossRefPubMedGoogle Scholar
  21. 21.
    Madlener S, Illmer C, Horvath Z et al (2007) Gallic acid inhibits ribonucleotide reductase and cyclooxygenases in human HL-60 promyelocytic leukemia cells. Cancer Lett 245:156–162CrossRefPubMedGoogle Scholar
  22. 22.
    Gali HU, Perchellet EM, Klish DS, Johnson JM, Perchellet JP (1992) Hydrolyzable tannins: potent inhibitors of hydroperoxide production and tumor promotion in mouse skin treated with 12-Otetradecanoylphorbol-13-acetate in vivo. Int J Cancer 51:425–432CrossRefPubMedGoogle Scholar
  23. 23.
    Gali HU, Perchellet EM, Perchellet JP (1991) Inhibition of tumor promoter- induced ornithine decarboxylase activity by tannic acid and other polyphenols in mouse epidermis in vivo. Cancer Res 51:2820–2825PubMedGoogle Scholar
  24. 24.
    Seo SY, Sharma VK, Sharma NJ (2003) Mushroom tyrosinase: Recent prospects. J Agric Food Chem 51:2837–2853CrossRefPubMedGoogle Scholar
  25. 25.
    Hsu CL, Yen GC (2007) Effect of gallic acid on high fat diet-induced dyslipidaemia, hepatosteatosis and oxidative stress in rats. Br J Nutr 98:727–735CrossRefPubMedGoogle Scholar
  26. 26.
    Ohkawa H, Ohisi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358CrossRefPubMedGoogle Scholar
  27. 27.
    Rao KS, Recknagel RO (1968) Early onset of lipid peroxidation in rat liver after carbon tetrachloride administration. Exp Mol Pathol 9:271–278CrossRefPubMedGoogle Scholar
  28. 28.
    Jiang ZY, Hunt JV, Wolff SP (1992) Ferrous ion oxidation in the presence of xylenol orange for detection of lipid hydroperoxides in low density lipoprotein. Anal Biochem 202:384–389CrossRefPubMedGoogle Scholar
  29. 29.
    Kakkar PS, Das B, Viswanathan PN (1984) A modified spectrophotometric assay of superoxide dismutase. Ind J Biochem Biophys 21:130–132Google Scholar
  30. 30.
    Sinha KA (1972) Colorimetric assay of catalase. Anal Biochem 47:389–394CrossRefPubMedGoogle Scholar
  31. 31.
    Ellman GL (1959) Tissue sulphydryl groups. Arch Biochem Biophys 82:72–77CrossRefGoogle Scholar
  32. 32.
    Carlberg I, Mannervik B (1985) Glutathione reductase. Methods in enzymology, vol. 7. Academic, New York, pp 484–490Google Scholar
  33. 33.
    Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG (1973) Selenium: biochemical role as a component of glutathione peroxidase. Science 179:588–590CrossRefPubMedGoogle Scholar
  34. 34.
    Roe JM, Kuether CA (1943) Detection of ascorbic acid in whole blood, and urine through 2,4-DNPH derivative of dehydroascorbic acid. J Biol Chem 147:399–407Google Scholar
  35. 35.
    Baker H, Frank O, DeAngelis B, Feingold S (1980) Plasma tocopherol in man at various times after ingesting free or acetylated tocopherol. Nutr Rep Int 21:531–536Google Scholar
  36. 36.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with Folin’s phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  37. 37.
    Saroja M, Balasenthil S, Nagini S (1999) Tissue lipid peroxidation and glutathione dependant enzyme status in patients with oral squamous cell carcinoma. Cell Biochem Funct 17:213–216CrossRefPubMedGoogle Scholar
  38. 38.
    Van Rossen ME, Sluiter W, Bonthuis F et al (2000) Scavenging of reactive oxygen species leads to diminished peritoneal tumor recurrence. Cancer Res 60:5625–5629PubMedGoogle Scholar
  39. 39.
    Fang YZ, Yang S, Wu G (2002) Free radicals, antioxidants, and nutrition. Nutrition 18:872–879CrossRefPubMedGoogle Scholar
  40. 40.
    Cheesman H, Collins H, Proudfoot K et al (1986) Studies on lipid peroxidation in normal and tumor tissues. J Biol Chem 235:507–514Google Scholar
  41. 41.
    Tanaka T (1997) Effect of diet on human carcinogenesis. Crit Rev Oncol Hematol 25(2):73–95CrossRefPubMedGoogle Scholar
  42. 42.
    Tanaka T, Kawabata K, Kakumoto M, Hara A, Murakami A, Kuki W, Takahashi Y, Yonei H, Maeda M, Ota T, Odashima S, Yamane T, Koshimizu K, Ohigashi H (1998) Citrus auraptene exerts dose-dependent chemopreventive activity in rat large bowel tumorigenesis: the inhibition correlates with suppression of cell proliferation and lipid peroxidation and with induction of phase II drug-metabolizing enzymes. Cancer Res 58:2550–2556PubMedGoogle Scholar
  43. 43.
    Pillai MG, Thampi BS, Menon VP, Leelamma S (1999) Influence of dietary fiber from coconut kernel (Cocos nucifera) on the 1,2-dimethylhydrazine-induced lipid peroxidation in rats. J Nutr Biochem 10:555–560CrossRefPubMedGoogle Scholar
  44. 44.
    Schmelz EM, Sullards MC, Dillehay DL, Merrill AH (2000) Colonic cell proliferation and aberrant crypt foci formation are inhibited by dairy glycospingolipids in 1,2-dimethylhydrazine-treated CFI mice. J Nutr 130:522–527PubMedGoogle Scholar
  45. 45.
    Nakagami K, Uchida T, Ohwada S, Koibuchi Y, Morishita Y (1999) Increased choline kinase activity in 1,2-dimethylhydrazine induced rat colon cancer. Jpn J Cancer Res 90:1212–1217PubMedGoogle Scholar
  46. 46.
    Vennila S, Karthik KV, Nalini N (2009) Effect of morin on tissue lipid peroxidation and antioxidant status in 1,2-dimethylhydrazine induced experimental colon carcinogenesis. Invest New Drugs 27:21–30, doi: 10.1007/s10637-008-9136-1 CrossRefGoogle Scholar
  47. 47.
    Aranganathan S, Panneerselvam J, Nalini N (2008) Hesperetin exerts dose dependent chemopreventive effect against 1,2-dimethyl hydrazine induced rat colon carcinogenesis. Invest New Drugs. doi: 10.1007/s10637-008-9158-8
  48. 48.
    Son S, Lewis BA (2002) Free radical scavenging and antioxidative activity of caffeic acid amide and ester analogues: structure activity relationship. J Agric Food Chem 50:468–472CrossRefPubMedGoogle Scholar
  49. 49.
    Lu Z, Nie G, Belton PS, Tang H, Zhao B (2006) Structure–activity relationship analysis of antioxidant ability and neuroprotective effect of gallic acid derivatives. Neurochem Int 48:263–274CrossRefPubMedGoogle Scholar
  50. 50.
    Burton GW, Cheesman KN, Ingold KV, Seater TF (1983) Lipid antioxidants and products of lipid peroxidation as potential tumor protective agents. Biochem Soc Trans 11:261–262PubMedGoogle Scholar
  51. 51.
    Rajeshkumar NV, Kuttan R (2003) Modulation of carcinogenic response and antioxidant enzymes of rats administered with 1,2- dimethylhydrazine by Picroliv. Cancer Lett 191:137–143CrossRefPubMedGoogle Scholar
  52. 52.
    Slaga TJ (1995) Inhibition of the induction of cancer by antioxidants. Adv Exp Med Biol 369:167–174PubMedGoogle Scholar
  53. 53.
    Hatano T, Edamatsu R, Hiramatsu M, Mori A, Fujita Y, Yasuhara T, Yoshida T, Okuda T (1989) Effects of tannins and related polyphenols on superoxide anion radical, and on, 1,1-diphenyl-2-picrylhydrazyl radical. Chem Pharm Bull 37:2016–2021Google Scholar
  54. 54.
    Michiels C, Raes M, Toussaint O, Remach J (1994) Importance of Se-glutathione, catalase, and Cu/Zn superoxide dismutase for cell survival against oxidative stress. Free Radic Biol Med 17:235–248CrossRefPubMedGoogle Scholar
  55. 55.
    Meister A (1974) Glutathione, metabolism and function via the gamma-glutamyl cycle. Life Sci 15:177–190CrossRefPubMedGoogle Scholar
  56. 56.
    Brigelius-Flohe R (1999) Tissue-specific functions of individual glutathione peroxidases. Free Radic Biol Med 27:951–965CrossRefPubMedGoogle Scholar
  57. 57.
    Slater TF, Bendetto C, Burton GW, Cheeseman KH, Ingold KG, Nodes JT (1984) Lipid peroxidation in animal tumors. A disturbance in the control of cell division. In: Thaler-Dao H, Crastes dePaulet A, Paolettiz R (eds) Eicosanoids and cancer. Raven, New York, p 21Google Scholar
  58. 58.
    Singh RP, Banerjee S, Kumar PVS, Raveesha KA, Rao AR (2006) Tinospora cordifolia induces enzymes of carcinogen/drug metabolism and antioxidant system, and inhibits lipid peroxidation in mice. Phytomedicine 13:74–84CrossRefPubMedGoogle Scholar
  59. 59.
    Kim YJ, No JK, Lee JH, Chung HY (2005) 4,4′-Dihydroxybiphenyl as a new potent tyrosinase inhibitor. Biol Pharm Bull 28:323–327CrossRefPubMedGoogle Scholar
  60. 60.
    Nerya O, Musa R, Khatib S, Tamir S, Vaya J (2004) Chalcones as potent tyrosinase inhibitors: the effect of hydroxyl positions and numbers. Phytochemistry 65:1389–1395CrossRefPubMedGoogle Scholar
  61. 61.
    Tanaka T, Kawabata K, Kakumoto M, Makita H, Ushida J, Honjo S, Hara A, Tsuda H, Mori H (1999) Modifying effects of a flavonoid morin on azoxymethane-induced large bowel tumorigenesis in rats. Carcinogenesis 20:1477–1484CrossRefPubMedGoogle Scholar
  62. 62.
    Bertram JS, Kolonel LN, Meyskens FL (1987) Rationale and strategies for chemoprevention of cancer in humans. Cancer Res 47:3012–3031PubMedGoogle Scholar
  63. 63.
    Mehlhorn RJ, Sumida S, Packer L (1989) Tocopheroxyl radical persistence and tocopherol consumption in liposomes and in vitamin E-enriched rat liver mitochondria and microsomes. J Biol Chem 264:13448–13452PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Jebakkan Senapathy Giftson
    • 1
  • Sathiavelu Jayanthi
    • 1
  • Namasivayam Nalini
    • 1
  1. 1.Department of Biochemistry & Biotechnology, Faculty of ScienceAnnamalai UniversityAnnamalainagarIndia

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