Indian Journal of Clinical Biochemistry

, Volume 26, Issue 3, pp 290–295

Chemoprotective Role of Triphala Against 1,2-Dimethylhydrazine Dihydrochloride Induced Carcinogenic Damage to Mouse Liver

Original Article


The present study was carried out to investigate the protective role of Triphala (a combination in equal proportions by weight of fruit powder of Terminalia belerica, Terminalia chebula and Emblica officinalis) against 1,2-dimethylhydrazinedihydrochloride (DMH) induced Endoplasmic reticulum stress (ER stress) in mouse liver. An oral dose of 3 mg/kg body wt in drinking water for 5 weeks significantly (P < 0.001) increased the levels of serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), serum Alkaline phosphatase (ALP) and total bilirubin thus suggesting damage to mouse liver and biliary dysfunction. The DMH administration invariably led to increase in the liver microsomal proteins of molecular weight of about 29 (ERp29) and 53 kDa (ERp53) and decrease in the protein of molecular weight of 36 kDa (ERp36) thereby suggesting the interference of DMH and its metabolites with normal protein biosynthesis and folding, in the reticular membranes of the liver cells thus developing ER stress. Histological studies show necrosis, large sized hepatocytes with increased N:C ratio, aberrant mitotic figures and prominent nucleoli in the liver of DMH treated mice. In animals fed 5% Triphala in diet (w/w) during DMH administration, there was significant decrease in the above changes in the liver suggesting the suppression of DMH induced ER stress in liver. Triphala significantly (P < 0.05) decreased lipid peroxidation and also the activity of lactate dehydrogenase (LDH) in mouse liver. It simultaneously increased the level of reduced glutathione (GSH) and the activity of glutathione-S-transferase (GST) thereby suggesting that it prevents peroxidative damage and also diverts the active metabolites (electrophiles) of DMH from their interactions with critical cellular bio-molecules which could be responsible for its protective action against DMH.


1,2-Dimethylhydrazinedihydrochloride Neoplastic lesions Triphala ER stress ERp29 ERp53 Antioxidant status Chemoprotective effect 


  1. 1.
    Toth B. Morphological studies of angiosarcinomas induced by 1,2-dimethylhydrazine in syrian golden hamsters. Cancer Res. 1972;32(12):2818–27.PubMedGoogle Scholar
  2. 2.
    Nomura K, Schlake W, Grundman E. New aspects of intestinal carcinogenesis by DMH and the influence of antilymphocyte globulin on it’s progress. Cancer Res. 1978;92(1):17–73.Google Scholar
  3. 3.
    Druckery H, Preusmann R, Schmahl D, Blum G. Topics in chemical carcinogenesis. In: Nakahara M, editor, Tokyo: University of Tokyo Press; 1972. p. 73–103.Google Scholar
  4. 4.
    Nagasawa HT, Shirota FN, Matsumoto H. Decomposition of methylazoxy methanol, the aglycone of cycansin in D2O. Nature. 1972;236:234–5.PubMedCrossRefGoogle Scholar
  5. 5.
    Comstok CC, Lawson LH, Greene EA, Oberst FW. AMA Arch Ind Health. 1954;10:476–9.Google Scholar
  6. 6.
    Hawks A, Magee PN. The alkylation of nucleic acids of rat and mouse in vivo by the carcinogen 1,2-dimethylhydrazine. Br J Cancer. 1974;30:440–7.PubMedCrossRefGoogle Scholar
  7. 7.
    Swenberg JA, Cooper HLN, Bucheler J, Kleihues P. 1,2-Dimethylhydrazine induced methylation of DNA bases in various rat organs and the effect of pre treatment with disulfiram. Cancer Res. 1979;32:146–52.Google Scholar
  8. 8.
    Sharma KK, Pathak RM, ShramaV, Dani HM. Effects of orally administered 1,2-dimethylhydrazine on lipid and protein composition of mouse small intestinal brush border membranes. Res Bull PU Chandigarh, India 1995;45(1–4): 1–9Google Scholar
  9. 9.
    Sharma KK, Sharma V, Dani HM. Effects of orally administered 1,2-Dimethylhydrazine on the absorptive activities of small intestine of mouse. Res Bull PU, Chandigarh, India 1995;45(I–IV): 11–17Google Scholar
  10. 10.
    Toth B. Synthetic and naturally occurring hydrazines as possible cancer causative agents. Cancer Res. 1975;35(12):3693–7.PubMedGoogle Scholar
  11. 11.
    Sharma KK. Biochemical studies on the structural and functional aspects of cell membranes during chemical carcinogenesis PhD thesis. Chandigarh: Panjab University; 1991. p. 5–7.Google Scholar
  12. 12.
    Liu YY. Chemical studies on tobacco smoke. quantitative analysis of hydrazine in tobacco and cigarette smoke. Anal Chem. 1974;46:885–9.PubMedCrossRefGoogle Scholar
  13. 13.
    Kostela JG, Lawrence BH. Hydrocarbon constituents from white strains of the mushroom Agaricus bisporus. J Agr Food Chem. 1981;20(1):185–6.CrossRefGoogle Scholar
  14. 14.
    Wilbert S, Steinbrecher K, Gunderson E. Prevalence of hydrazine derivatives in food and food products. J Agr Food Chem. 1990;52:214.Google Scholar
  15. 15.
    Parthasarthy R, Raghupati Sarma G, Janardhanam B, Ramachandran P, Santha T, Sibasubramaniam S. Hepatic toxicity in South Indian patients during treatment of tuberculosis with short-course regimens containing isoniazide, aflatoxin and pyrazinamide. Tubercle. 1986;67:99–106.CrossRefGoogle Scholar
  16. 16.
    Shakun NP, Tabachuk OP. The comparative action of isoniazide, aflatoxin and ethambutol on liver function. Eksp Klin Farmakol. 1992;55:45.Google Scholar
  17. 17.
    Suvarajan VV. Ayurvedic drugs and their plant sources. Lebanon, NH: International Science Publisher; 1994.Google Scholar
  18. 18.
    Suresh K, Vasudevan DM. Augmentation of murine natural killer cell and antibody dependent cellular cytotoxicity activities by Phyllanthus emblica, a new immuno modulator. J Ethnopharmacol. 1994;44:55–60.PubMedCrossRefGoogle Scholar
  19. 19.
    Sabu MC, Kuttan R. Anti-diabetic activity of medicinal plants and it’s relationship with their antioxidant property. J Ethno pharmacol. 2002;81(2):155–62.Google Scholar
  20. 20.
    Tokura K, Kagawa S. Anti cancer agents containing chebulanin from Terminalia chebula. Jpn Kokai Tokyo Koho JP. 1995;7:138–65.Google Scholar
  21. 21.
    Kaur S, Arora S, Kaur K, Kumar S. The in vitro antimutagenic activity of Triphala—an Indian herbal drug. Food Chem Toxicol. 2002;40(4):527–34.PubMedCrossRefGoogle Scholar
  22. 22.
    Jagetia GC, Baliga MS, Malagi KJ, Kamath MS. The evaluation of the protective effect of Triphala (an ayurvedic rejuvenating drug) in mice exposed to y radiation. Phytomedicine. 2002;9:99–108.PubMedCrossRefGoogle Scholar
  23. 23.
    Reddy VRC. Cardiotonic activity of the fruits of Terminalia chebula. Fitoterapia. 1990;61:517–25.Google Scholar
  24. 24.
    Ghosal S, Tripathi VK, Chauhan S. Active constituents of Emblica officinalis, Part I. The chemistry and antioxidative effects of two new hydrolysable tannins, emblicanin a and b. Ind J Chem Section B-Organic Chemistry including Medicinal Chemistry. 1996;35:941–8.Google Scholar
  25. 25.
    Bhattacharya A. Antioxidant activity of active tannoid principles of Emblica officinalis (amla). Ind J Expt Biol. 1999;37:676–80.Google Scholar
  26. 26.
    Deep G, Dhiman M, Rao AR, Kale RK. Chemopreventive potential of Triphala (s composite Indian drug) on Benzo (a) Pyrene induced Forestomach Tumorigenesis in murine tumor model. J Exp Clin Cancer Res. 2005;24(4):555–63.PubMedGoogle Scholar
  27. 27.
    Bedell MA, Lewis JG, Billing KC, Swenberg JA. Cell specificity in hepatocarcinogenesis: preferential accumulation of O6 methylguanine in target cell DNA during continuous exposure to rats to 1,2-dimethylhydrazine. Cancer Res. 1982;42(8):3079–83.PubMedGoogle Scholar
  28. 28.
    Reitman S, Frankel S. A colorimetric method for the determination of serum glutamic oxaloacetic and glutamic pyruvic transaminases. Am J Clin Pathol. 1957;28:53–6.Google Scholar
  29. 29.
    Kind PRN, King EJJ. Estimation of plasma phosphatase by determination of hydrolyzed phenol with anti-pyrine. J Clin Pathol. 1957;7:322–30.CrossRefGoogle Scholar
  30. 30.
    Malloy HT, Evelyn KA. The determination of bilirubin with the photometric colorimeter. J Biol Chem. 1937;119:481–90.Google Scholar
  31. 31.
    Moron MA, Depierre JW, Mannervick B. Levels of glutathione, glutathione reductase and glutathione-S-transferase activities in rat lung and liver. Biochim Biophys Acta. 1979;582:67–78.PubMedGoogle Scholar
  32. 32.
    Mihira M, Uchiyama M. Determination of malonaldehyde precursors in tissues by thiobarbituric acid test. Anal Biochem. 1978;86:271–8.CrossRefGoogle Scholar
  33. 33.
    Habig WH, Pabst MJ, Jokoby WB. The first step in mercapturic acid formation. J Biol Chem. 1994;249:7130–9.Google Scholar
  34. 34.
    Bergmeyer HU, Bernt E. Methods in enzymatic analysis, vol. II. New York and London: Verlag Academic Pres; 1974. p. 5574–9.Google Scholar
  35. 35.
    Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680–5.PubMedCrossRefGoogle Scholar
  36. 36.
    Lowry OH, Rosenbrough NJ, Farr A, Randalll RJ. Protein measurement with the folin phenol reagent. J Biol Chem. 1951;193:265–75.PubMedGoogle Scholar
  37. 37.
    Harper HA. The functions and tests of liver, In: Review of physiological chemistry. Los Altos, California: Lange Medical Publishers; 1961. p. 271–283.Google Scholar
  38. 38.
    Sathiyanaryanan L, Arulmozhi S, Chdiambarnathan N. Anticholesterlemic, hepatoprotective and antioxidant activity of Gilnus lotoides Linn. against ethanol induced liver damage in rats. Phcog Mag. 2006;2:160–2.Google Scholar
  39. 39.
    Dortman RB, Lawhorn GT. Serum enzymes as indicators of chemical induced liver damage. Drug Chem Toxicol. 1978;1:163–71.CrossRefGoogle Scholar
  40. 40.
    Abul K, Najmi KK, Pillai SN, Aqil PM. Free radical scavenging and hepatoprotective activity of jigrine against galactosamine induced liver damage. Drug Chem Toxicol. 1978;1:163–71.CrossRefGoogle Scholar
  41. 41.
    Roa RR. Mechanism of drug induced hepatotoxicity. Ind J Pharamcol. 1973;5:313–8.Google Scholar
  42. 42.
    Kim MK, Hyun SH, Choung SY. Effect of herbal extract mixtures on serum and liver lipid metabolism in chronic ethanol administered rats. J Health Sci. 2006;52:344–51.CrossRefGoogle Scholar
  43. 43.
    Suresh Kumar SV, Sujatha C, Syamala J, Nagasudha B, Mishra SH. Protective effect of root extract of operculina terpethum Linn. Against paracetamol induced hepatotoxicity in rats. Ind J Pharma Sci. 2006;68:32–5.CrossRefGoogle Scholar
  44. 44.
    Singh B, Saxena AK, Chandan BK, Anand KK, Suri OP, Suri KA, Satti NK. Hepatoprotective activity of verbenalin on experimental liver damage in rodents. Fitoterapia. 1989;69:135–40.Google Scholar
  45. 45.
    Meshkibaf MH, Ebeahimi A, Ghodsi R, Ahmadi A. Chronic effects of lamotrigine on liver function in adult male rats. Ind J Clin Biochem. 2006;21:161–264.CrossRefGoogle Scholar
  46. 46.
    Hainaut P, Hollstein M. p53 and human cancer: the first ten thousand mutations. Adv Cancer Res. 2000;77:81–137.PubMedCrossRefGoogle Scholar
  47. 47.
    Zhang D, Richardson DR. Endoplasmic reticulum protein29 (ERp29): an emerging role in cancer. Int J Biochem Cell Biol. 2011;43(1):33–6.PubMedCrossRefGoogle Scholar
  48. 48.
    Ames BN, Shigenoga MK, Hagen TM. Oxidants, antioxidants and degenerative diseases of ageing. Proc Nat Acad Sci USA. 1993;90:7915–22.PubMedCrossRefGoogle Scholar
  49. 49.
    Ketterer B. Protective role of glutathione and glutathione-S-transferases in mutagenesis and carcinogenesis. Mutat Res. 1988;202:343.PubMedGoogle Scholar
  50. 50.
    Aggarwal A, Choudhary D, Uperti M. Rath:II studies in liver as a distant organ of tumour bearing mice. Mol Cell Biochem. 2001;224:9–17.CrossRefGoogle Scholar
  51. 51.
    Dixon DP, Cummins I, Cle DJ, Edwards R. Glutathione mediated detoxification system in plants. Curr Opin Plant Biol. 1998;1:256.Google Scholar
  52. 52.
    Talalay P, Delong MJ, Prochaska HJ. Molecular mechanism in protection against carcinogensis. In: Cory JG, Szentivani Plenum A, editors. Cancer biology and therapeutics. New York: Plenum Press; 1987. p. 187.Google Scholar

Copyright information

© Association of Clinical Biochemists of India 2011

Authors and Affiliations

  1. 1.Department of Biochemistry and MicrobiologyDr Rajendra Prasad Govt Medical College Kangra at TandaHimachal PradeshIndia

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