ANTIOXIDANT ACTIVITY OF TRADITIONAL SIDDHA FORMULATION ON CCL4 INDUCED LIVER FIBROSIS IN RATS
Objective: The main objective of this study was to evaluate the antioxidant activity of Traditional Siddha Formulation (TSF) on CCl4 induced liver fibrosis in rats.
Methods: In this study, plant materials were collected, shade dried, mixed in equal proportion and extraction process was done to prepare TSF. Liver injury was induced by intraperitoneal injection of 1 ml/kg body weight of both CCl4 and olive oil (2:3 v/v) mixture weekly twice for 8 w. The levels of thiobarbituric acid reactive substances (TBARS), lipid peroxides (LPO), protein carbonyl (PC), superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx), glutathione reductase (GR), Vitamin C (VIT C), Vitamin E (VIT E), GSH and Total thiols (TTS) were measured in the liver of experimental rats. Histopathological changes in the liver of experimental rats were assessed for each group using hematoxylin and eosin
Results: At the end of the 8-week experimental period, histopathological examination was demonstrated which indicates TSF could attenuate the inflammation and reduced the score of liver fibrosis. The administration of TSF significantly decreased the levels of TBARS (4.11Â±0.09, pË‚0.01), LPO (53.15Â±0.79, pË‚0.01), PC (7.31Â±0.26, pË‚0.01) and significantly increased the levels of antioxidant enzymes such as SOD (7.10Â±0.16, pË‚0.01), CAT (62.14Â±1.61, pË‚0.01), GPx (90.79Â±1.05, pË‚0.01), GR (144.06Â±1.61, pË‚0.01), GSH (35.24Â±0.82, pË‚0.01), VIT C (3.29Â±0.10, pË‚0.01), VIT E (2.42Â±0.14, pË‚0.01) and TTS (15.15Â±0.21, pË‚0.01).
Conclusion: TSF inhibits CCl4 intoxicated hepatic fibrosis in Wistar Albino rats which may be due to the action of synergistically active phytochemicals present in the formulation. Enzymatic and non-enzymatic antioxidant enzyme levels were found to be increased in the treatment group which may be related to the therapeutic effect of TSF.
2. Abdelaziz DHA, Ali SA. The protective effect of Phoenix dactylifera L. seeds against CCl4 induced hepatotoxicity in rats. J Ethnopharmacol 2014;155:736â€“43.
3. Fujii T, Fuchs BC, Yamada S, Lauwers GY, Kulu Y, Goodwin JM, et al. Mouse model of carbon tetrachloride induced liver fibrosis: histopathological changes and expression of CD133 and epidermal growth factor. BMC Gastroenterol 2010;10:79.
4. Weiler-Normann C, Herkel J, Lohse AW. Mouse models of liver fibrosis. Z Gastroenterol 2007;45:43-50.
5. Iwaisako K, Brenner DA, Kisseleva T. Whatâ€™s new in liver fibrosis? The origin of myofibroblasts in liver fibrosis. J Gastroenterol Hepatol 2012;27:65â€“8.
6. Sanchez-Valle V, Chavez-Tapia NC, Uribe M, Mendez-Sanchez N. Role of oxidative stress and molecular changes in liver ï¬brosis: a review. Curr Med Chem 2012;19:4850â€“60.
7. Sri kamatchi Priya Ramamoorthy, Diwakar Manickam, Samu Subramaniam, Shyama Subramaniam. Standardization and phytochemical screening of traditional formulation. Int J Curr Pharm Res 2017;9:70-4.
8. Sri Kamatchi Priya Ramamoorthy, Diwakar Manickam, Santhosh Kumar, Samu Subramaniam, Shyama Subramaniam. In vitro antioxidant and cytotoxicity analysis of traditional formulation. J Chem Pharm Res 2017;9:315-25.
9. Diwakar Manickam, Sri Kamatchi Priya Ramamoorthy, B Santhosh Kumar, Samu Subramaniam, Shyama Subramaniam. Biochemical and histopathological changes of traditional siddha formulation on CCl4 induced liver fibrosis in rats. Int J Pharm Pharm Sci 2017;9:120-3.
10. F Malekzadeh, H Ehsanifar, M Shahamat, M Levin, RR Colwell. Antibacterial activity of black myrobalan (Terminalia chebula retz) against Helicobacter pylori. Int J Antimicrob Agents 2001;18:85â€“8.
11. HY Cheng, TC Lin, KH Yu, CM Yang, CC Lin. Antioxidant and free radical scavenging activities of Terminalia chebula. Biol Pharm Bull 2003;26:1331â€“5.
12. R Karuna, S Sreenivasa Reddy, R Baskar, D Saralakumari. Antioxidant potential of aqueous extract of Phyllanthus amarus in rats. Indian J Pharmacol 2009;41:64â€“7.
13. Vaidya Ratnam KS, Murugesa Mudaliar. Gunapadam. (Siddha Materia Medica) 1 edition; 1936. p. 229.
14. Vijayastelter B Liju, Kottarapat Jeena, Ramadasan Kuttan. An evaluation of antioxidant, anti-inflammatory, and antinociceptive activities of essential oil from Curcuma longa. L. Indian J Pharmacol 2011;43:526â€“31.
15. Mnif S, Aifa S. Cumin (Cuminum cyminum L.) from traditional uses to potential biomedical applications. Chem Biodivers 2015;12:733-42.
16. Khosla S, Sharma S. A short description on pharmacogenetic properties of Emblica officinalis. Spatula DD 2012;2:187â€“93.
17. Sabu MC, Kuttan R. Antidiabetic and antioxidant activity of Terminalia belerica. Roxb. Indian J Exp Biol 2009;47:270-5.
18. Finkel T, Holbrook NJ. Oxidants, oxidative stress and the biology of ageing. Nature 2000;408:239â€“47.
19. Apel K, Hirt H. Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Ann Rev Plant Biol 2004;55:373â€“99.
20. Medina J, Moreno-Otero R. Pathophysiological basis for antioxidant therapy in chronic liver disease. Drugs 2005;65:2445â€“61.
21. Muriel P. Role of free radicals in liver diseases. Hepatol Int 2009;3:526â€“36.
22. HE Seifried, SS McDonald, DE Anderson, P Greenwaldand JA Milner. The antioxidant conundrum in cancer. Cancer Res 2003;63:4295â€“8.
23. H Ohkawa, N Ohishi, K Yagi. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95:351â€“8.
24. TPA. Devesagayam and U. Tarachand. Decreased lipid peroxidation in the rat kidney during gestation. Biochem Biophys Res Commun 1987;145:134â€“8.
25. Z Reznick, L Packer. Oxidative damage to proteins: spectrophotometric method for carbonyl assay. Methods Enzymol 1994;233:357â€“63.
26. K Sinha. Colorimetric assay of catalase. Anal Biochem 1972;47:389â€“94.
27. Misra HP, Fridovich I. The role of superoxide anion in the antioxidation of epi-epinephrine and a simple assay for superoxide dismutase. J Biol Chem 1972;247:3170â€“5.
28. JT Rotruck, AL Pope, HE Ganther, AB Swanson, DG Hafeman, WG Hoekstra. Selenium: biochemical role as a component of glutathione peroxidase. Science 1973;179:588â€“90.
29. MS Moron, JW Depierre, B Mannervik. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 1979;582:67â€“78.
30. ML Quaife, Dju MY. Chemical estimation of vitamin E in tissue and the tocopherol content of some normal human tissues. J Biol Chem 1949;80:263â€“72.
31. ST Omaye, JD Turnbull, HE Sauberlich. Selected methods for the determination of ascorbic acid in animal cells, tissues, and ï¬‚uids. Methods Enzymol 1979;62:3â€“11.
32. Lindenmaier H, Becker M, Haefeli WE, Weiss J. Interaction of progestins with the human multidrug resistance-associated protein 2 (MRP2). Drug Metab Dispos 2005;33:1576â€“9.
33. Williams T, Burk RF. Carbon tetrachloride hepatotoxicity: an example of free radical-mediated injury. Semin Liver Dis 1990;10:279â€“84.
34. Vuda M, Dâ€™Souza R, Upadhya S, Kumar V, Rao N, Kumar V, et al. Hepatoprotective and antioxidant activity of aqueous extract of Hybanthus enneaspermus against CCl4-induced liver injury in rats. Exp Toxicol Pathol 2012;64:855â€“5.
35. Tao Cong, Xue-Yuan Jin, Lin Zhao, Long Ma, Rui-Sheng Li, Ping Zhao, et al. Anti-fibrotic effects of the masson pine pollen aqueous extract on hepatic fibrosis rat model. Int J Clin Exp Pathol 2015;8:4651-61.
36. Bedossa P. Stimulation of collagen a1 (I) gene expression is associated with lipid peroxidation in hepatocellular injury: a link to tissue ï¬brosis. Hepatology 1994;19:1262â€“71.
37. Abbas AT, El-Shitany NA, Shaala LA, Ali SS, Azhar EI, Abdel-Dayem UA, et al. Red sea Suberea mollis sponge extract protects against CCl4-induced acute liver injury in rats via an antioxidant mechanism. Evid-Based Complementary Altern Med 2014;745606. http://dx.doi.org/10.1155/2014/745606
38. Sundari PN, Wilfred G, Ramakrishna B. Does oxidative protein damage play a role in the pathogenesis of carbon tetrachloride-induced liver injury in the rat. Biochim Biophys Acta1 997;1362:169-76.
39. V Reutrakul, N Ningnuek, M Pohmakotr. Anti HIV-1ï¬‚avonoid glycosides from Ochnaintegerrima. Planta Med 2007;73:683â€“8.
40. Jalali Ghassam B, Ghaffari H, Prakash HS, Kini KR. Antioxidant and hepatoprotective effects of Solanum xanthocarpum leaf extracts against CCl4-induced liver injury in rats. Pharm Biol 2014;52:1060â€“8.
41. Reiter RJ, Tan DX, Osuna C, Gitto E. Actions of melatonin in the reduction of oxidative stress. A review. J Biomed Sci 2000;7:444â€“58.
42. Hsu YW, Tsai CF, Chang WH, Ho YC, Chen WK, Lu FJ. Protective effects of Dunaliella salinaâ€”a carotenoids-rich alga, against carbon tetrachloride-induced hepatotoxicity in mice. Food Chem Toxicol 2008;46:3311â€“7.
43. Li S, Li SK, Gan RY, Song FL, Kuang L, Li HB. Antioxidant capacities and total phenolic contents of infusions from 223 medicinal plants. Ind Crops Prod 2013;51;289â€“98.
44. H Glatt, T Friedberg, PL Grover. Inactivation of adiol-epoxide and a K-region epoxide with high eï¬ƒciency by glutathione transferase X. Cancer Res 1983;43:5713â€“7.
45. T Byers, G Perry. Dietary carotenes, vitamin C, and vitamin E as protective antioxidants in human cancers. Annu Rev Nutr 1992;12:139â€“59.
46. Bhattacharya A Chatterjee, S Ghosal, SK Bhattacharya. Antioxidant activity of active tannoid principles of Emblica oï¬ƒcinalis (amla). Indian J Exp Biol 1999;37:676â€“80.