DEVELOPMENT OF CARBON TETRACHLORIDE-INDUCED CHRONIC HEPATOTOXICITY MODEL IN RATS AND ITS APPLICATION IN EVALUATION OF HEPATOPROTECTIVE ACTIVITY OF SILYMARIN

  • Gopi H Shah Department of Pharmacology and Toxicology, Ramanbhai Patel College of Pharmacy, Charotar University of Science and Technology, CHARUSAT – Campus, Changa, Petlad, Anand - 388 421, Gujarat, India
  • Bharat G Patel Department of Pharmacology, Charotar University of Science and Technology, CHARUSAT – Campus, Changa, Petlad, Anand - 388 421, Gujarat, India.
  • Gaurang B Shah Department of Pharmacology and Toxicology, K.B.Institute of Pharmaceutical Education and Research, Sector-23, GH-6, Gandhinagar, Gujarat, India

Abstract

Objective: The objective of the present work is to develop carbon tetrachloride (CCl4)-induced chronic hepatotoxicity model in rats and its applicationin evaluation of hepatoprotective activity of silymarin.

Methods: Animals were divided into four groups. Three groups were the disease induction group and 4th was the treatment group. In disease inductiongroups, chronic liver injury was induced by administration of CCl4 through intraperitoneal route (1 ml/kg) for 7-8 weeks. For treatment Group, 1 mlsilymarin suspension (orally) and CCl4 was given for 7-8 weeks. During disease induction and treatment period (7-8 weeks), blood samples werecollected and serum was separated which in turn used to analyze liver function tests such as serum glutamate oxaloacetate transaminase (SGOT),serum glutamate pyruvate transaminase (SGPT), alkaline phosphate (ALP), direct bilirubin, total protein (TP), and albumin (Alb) levels. Along withliver functional tests, tests to check cholesterol, glucose, and malondialdehyde (MDA) were also performed. Liver fibrosis and cirrhosis was quantifiedby histopathological studies of small portion of the excised liver. Model was validated by repetition of the experiment. Intermediate dissection wascarried out to measure an extent of liver damage.

Result: Serum SGOT, SGPT, ALP, and direct bilirubin were found to be significantly higher in CCl4 intoxicated rats. TP and Alb were decreased, andMDA was found to be significantly higher in CCl4 intoxicated rats, which is the main end product of lipid peroxidation. Whereas in the treatment groupsilymarin improved the liver functions in CCl4 toxicated drug.

Conclusion: We conclude that protein oxidation may play a role in the pathogenesis of CCl4 induced liver injury. The accumulation of oxidized proteinsmay be an early indication of CCl4 induced liver damage and silymarin found to be effective in liver injury by inhibiting protein oxidation.

Keywords: Liver fibrosis, Free radicals, Lipid peroxidation, Oxidative stress, Carbon tetrachloride, Liver biomarkers.

Author Biography

Gopi H Shah, Department of Pharmacology and Toxicology, Ramanbhai Patel College of Pharmacy, Charotar University of Science and Technology, CHARUSAT – Campus, Changa, Petlad, Anand - 388 421, Gujarat, India
Department of Pharmacology and toxicology

References

1. Banrida W, Juliane IB, Heather BC, Bellis JH, Cameron FK, Craig JM, et al. Toxicant-associated steatohepatitis. Toxicol Pathol 2013;41:343-60.
2. Cave M, Falkner KC, McClain CJ. Occupational and environmental liver disease. In: Zakim and Boyer’s Hepatology: A Textbook of Liver Disease. Philadelphia, PA: Elsevier Saunders; 2011. p. 476-92.
3. Friedman SL. Liver fibrosis. J Hepatol 2003;38(1):S38-53.
4. Bigoniya P, Singh CS, Shukla A. A comprehensive review of different liver toxicants used in experimental pharmacology. Int J Pharm Sci Drug Res 2009;1(3):124-35.
5. Renner H. The limited relevance of models used for testing human hepatic diseases and their prevention. In: Keppler E, Popper H, Bianchi L, Reutter W, editors. Mechanisms of Hepatocyte Injury and Death. Lancaster: MTP Press Ltd.; 1985. p. 311-20.
6. Cotran RS, Kumar V, Robbins SL. Genetic disorders. In: Cotran RS, Kumar V, Robbins SL, editors. Pathologic Basis of Disease. Vol. 5. Philadelphia, PA: WB Saunders Co.; 1994. p. 143.
7. Kaplowitz N, Aw TY, Simon FR, Stolz A. Drug-induced hepatotoxicity. Ann Intern Med 1986;104(6):826-39.
8. Kanchana N, Sadiq AM. Hepatoprotective effect of Plumbago zeylanica on paracetamol induced liver toxicity in rats. Int J Pharm Pharm Sci 2011;3:151-4.
9. Muriel P, Garciapiña T, Perez-Alvarez V, Mourelle M. Silymarin protects against paracetamol-induced lipid peroxidation and liver damage. J Appl Toxicol 1992;12(6):439-42.
10. Wagner H. Plant constituents with antihepatotoxic activity. In: Beal JL, Reinhard E, editors. Natural Products as Medicinal Agents. Stuttgart: Hippokrates-Verlag; 1981.
11. Bosisio E, Benelli C, Pirola O. Effect of the flavanolignans of Silybum marianum L. on lipid peroxidation in rat liver microsomes and freshly isolated hepatocytes. Pharmacol Res 1992;25(2):147-54.
12. Baer-Dubowska W, Szaefer H, Krajka-Kuzniak V. Inhibition of murine hepatic cytochrome P450 activities by natural and synthetic phenolic compounds. Xenobiotica 1998;28(8):735-43.
13. Campos R, Garrido A, Guerra R, Valenzuela A. Silybin dihemisuccinate protects against glutathione depletion and lipid peroxidation induced by acetaminophen on rat liver. Planta Med 1989;55(5):417-9.
14. Sandhu NS, Chopra D, Kaur S. Amelioration of paracetamol induced hepatotoxicity by a protein isolated from the leaves of the herb Cajanus Acutifolius Linn. Int J Pharm Pharm Sci 2010;2(3):75-80.
15. Sonnenbichler J, Zetl I. Biochemical effects of the flavanolignane silibinin on RNA, protein and DNA synthesis in rat livers. In: Cody V, Middleton E, Harbourne JB, editors. Plant Flavonoids in Biology and Medicine: Biochemical, Pharmacological, and Structure-Activity Relationships. New York, NY: Alan R. Liss; 1986. p. 319-31.
16. Fuchs EC, Weyhenmeyer R, Weiner OH. Effects of silibinim and of a synthetic analogue on isolated rat hepatic stellate cells and myofibroblasts. Arzneimittelforschung 1997;26:643-9.
17. Khan AA, Alzohairy M. Hepatoprotective effects of camel milk against CCl4-induced hepatotoxicity in Rats. Asian J Biochem 2011;6(2):171-80.
18. Larrey D. Drug-induced liver diseases. J Hepatol 2000;32 1 Suppl:77-88.
19. Liedtke C, Luedde T, Sauerbruch T, Scholten D, Streetz K, Tacke F, et al. Experimental liver fibrosis research: Update on animal models, legal issues and translational aspects. Fibrogenesis Tissue Repair 2013;6:19.
20. Nakano A, Kanda T, Abe H. Bone changes and mineral metabolism disorders in rats with experimental liver cirrhosis. J Gastroenterol Hepatol 1996;11:1143-54.
21. Karakus E, Karadeniz A, Simsek N, Can I, Kara A, Yildirim S, et al. Protective effect of Panax ginseng against serum biochemical changes and apoptosis in liver of rats treated with carbon tetrachloride (CCl4). J Hazard Mater 2011;195:208-13.
22. Stoyanovsky DA, Cederbaum AI. Metabolism of carbon tetrachloride to trichloromethyl radical: An ESR and HPLC-EC study. Chem Res Toxicol 1999;12(8):730-6.
23. Ramadori G, Saile B. Portal tract fibrogenesis in the liver. Lab Invest 2004;84:153-9.
24. Saile B, Ramadori G. Inflammation, damage repair and liver fibrosis - Role of cytokines and different cell types. Z Gastroenterol 2007;45(1):77-86.
25. Yu C, Wang F, Jin C, Wu X, Chan WK, McKeehan WL. Increased carbon tetrachloride-induced liver injury and fibrosis in FGFR4- deficient mice. Am J Pathol 2002;161(6):2003-10.
26. Kaplan MM. Laboratory tests. In: Schiff L, Schiff ER, editors. Diseases of the Liver. Vol. 7. Philadelphia, PA: JB Lippincott; 1993. p. 108-44.
27. Brent JA, Rumack BH. Role of free radicals in toxic hepatic injury. II. Are free radicals the cause of toxin-induced liver injury? J Toxicol Clin Toxicol 1993;31(1):173-96.
28. Zimmerman HJ, Kodera Y, West M. Effects of carbon tetrachloride poisoning on the plasma levels of cytoplasmic and mitochondrial enzymes in animals with nutritional fatty metamorphosis. J Lab Clin Med 1965;66:324-33.
29. Mehmetçik G, Ozdemirler G, Koçak-Toker N, Cevikbas U, Uysal M. Role of carnosine in preventing thioacetamide-induced liver injury in the rat. Peptides 2008;29(3):425-9.
30. Arıcı OF, Cetin N. Protective role of ghrelin against carbon tetrachloride (CCl4 induced coagulation disturbances in rats. Regul Pept 2011;166:139-42.
31. Binita S, Mehul C, Vandit T. Anti-fibrotic effect of heparin, silymarin and its combination on liver fibrosis model in rats. J Pharm Res Opin 2011;6:180-6.
32. Terblanche J, Hickman R. Animal models of fulminant hepatic failure. Dig Dis Sci 1991;36:770-4.
33. Mayer KE, Myers RP, Lee SS. Silymarin treatment of viral hepatitis: A systematic review. J Viral Hepat 2005;12:559-67.
34. Goudar MA, Jaydevappa H, Mahadevan KM, Shastry RA, Habbu PV, Sayeswar HA, et al. Isolation and characterization of secondary metabolite from habenaeria intermedia D. DON for screening hepatoprotective potential against carbon tetrachloride induced toxicity in albino rat liver. Int J Curr Pharm Res 2015;7(1):57-61.
35. Pradhan SC, Girish C. Hepatoprotective herbal drug, silymarin from experimental pharmacology to clinical medicine. Indian J Med Res 2006;124(5):491-504.
Statistics
296 Views | 640 Downloads
Citatons
How to Cite
Shah, G. H., B. G. Patel, and G. B. Shah. “DEVELOPMENT OF CARBON TETRACHLORIDE-INDUCED CHRONIC HEPATOTOXICITY MODEL IN RATS AND ITS APPLICATION IN EVALUATION OF HEPATOPROTECTIVE ACTIVITY OF SILYMARIN”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 10, no. 8, Aug. 2017, pp. 274-8, doi:10.22159/ajpcr.2017.v10i8.18701.
Section
Original Article(s)