The preventive effect of atorvastatin on liver fibrosis in the bile duct ligation rats via antioxidant activity and down-regulation of Rac1 and NOX1

Document Type : Original Article


1 Department of Biochemistry, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran

2 Department of Pharmacology, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran


Objective(s): Atorvastatin is a cholesterol-lowering agent capable of inhibiting 3-hydroxy-3-methylglutaryl coenzyme A reductase. Recent studies have demonstrated new facets of atorvastatin, such as antioxidant and anti-fibrotic properties. We investigated the effect of atorvastatin on hepatic injury via the measurement of the antioxidant capacity and protein expression of NOX1, Rac1-GTP, and Rac1 in a rat biliary duct ligation (BDL) model.
Materials and Methods: This study is regarded as experimental interventional research in which a total of 32 adult male Wistar rats (200-250 g) were assigned to 4 groups (eight rats per group) as follows: Control group; Control + At group (15 mg\kg\day atorvastatin); BDL group, and BDL+ At group (15 mg\kg\day atorvastatin). Expression levels of Rac1, NOX1, and Rac1-GTP were determined by western blot analysis. Besides, specific biomarkers of oxidative stress in hepatic tissues of all animals were also analyzed.
Results: Atorvastatin reduced liver injury via a decrease in the expression of NOX1, Rac1-GTP, and Rac1 in the BDL group (P<0.05), while the increased contents of protein thiol groups were observed, and the protein carbonylation was decreased in atorvastatin-treated BDL rats compared to the BDL group (P<0.05). Also, administration of atorvastatin in the BDL group significantly lowered oxidative stress through increasing the activity of catalase and superoxide dismutase in comparison with the BDL group (P<0.05).
Conclusion: It seems that atorvastatin has potential advantages in mitigation of liver fibrosis by a decrease in the expression of NOX1, Rac1-GTP, and Rac1, along with, a reduction in oxidative stress of liver tissues in rats induced by BDL. 


1. Zhen YZ, Li NR, He HW, Zhao SS, Zhang GL, Hao XF, et al. Protective effect of bicyclol against bile duct ligation-induced hepatic fibrosis in rats. World J Gastroenterol 2015; 21:7155-7164.
2. Klein S, Klosel J, Schierwagen R, Korner C, Granzow M, Huss S, et al. Atorvastatin inhibits proliferation and apoptosis, but induces senescence in hepatic myofibroblasts and thereby attenuates hepatic fibrosis in rats. Lab Invest 2012; 92:1440-1450.
3. Zhang F, Jin H, Wu L, Shao J, Zhu X, Chen A, et al. Diallyl Trisulfide Suppresses Oxidative Stress-Induced Activation of Hepatic Stellate Cells through Production of Hydrogen Sulfide. Oxid Med Cell Longev 2017; 2017:1406726.
4. Cui W, Matsuno K, Iwata K, Ibi M, Katsuyama M, Kakehi T, et al. NADPH oxidase isoforms and anti-hypertensive effects of atorvastatin demonstrated in two animal models. J Pharmacol Sci 2009; 111:260-268.
5. Liang S, Kisseleva T, Brenner DA. The Role of NADPH Oxidases (NOXs) in Liver Fibrosis and the Activation of Myofibroblasts. Front Physiol 2016; 7:17.
6. Aoyama T, Paik YH, Watanabe S, Laleu B, Gaggini F, Fioraso-Cartier L, et al. Nicotinamide adenine dinucleotide phosphate oxidase in experimental liver fibrosis: GKT137831 as a novel potential therapeutic agent. Hepatology 2012; 56:2316-2327.
7. Choi SS, Witek RP, Yang L, Omenetti A, Syn WK, Moylan CA, et al. Activation of Rac1 promotes the hedgehog-mediated acquisition of the myofibroblastic phenotype in rat and human hepatic stellate cells. Hepatology 2010; 52:278-290.
8. Boureux A, Vignal E, Faure S, Fort P. Evolution of the Rho family of Ras-like GTPases in eukaryotes. Mol Biol Evol 2007; 24:203-216.
9. Simon TG, King LY, Zheng H, Chung RT. Statin use is associated with a reduced risk of fibrosis progression in chronic hepatitis C. J Hepatol 2015; 62:18-23.
10. Nikolic T, Zivkovic V, Srejovic I, Stojic I, Jeremic N, Jeremic J, et al. Effects of atorvastatin and simvastatin on oxidative stress in diet-induced hyperhomocysteinemia in Wistar albino rats: a comparative study. Mol Cell Biochem 2017.
11. Kabirifar R, Rezaifar A, Binesh F, Bamdad K, Moradi A. Curcumin, quercetin and atorvastatin protected against the hepatic fibrosis by activating AMP-activated protein kinase. J Funct Foods 2018; 40:341-348.
12. Kilit C, Kocak FE, Pasali Kilit T. Comparison of the effects of high-dose atorvastatin and high-dose rosuvastatin on oxidative stress in patients with acute myocardial infarction: A pilot study. Turk Kardiyol Dern Ars 2017; 45:235-243.
13. Haddadian Z, Eftekhari G, Mazloom R, Jazaeri F, Dehpour AR, Mani AR. Effect of endotoxin on heart rate dynamics in rats with cirrhosis. Auton Neurosci 2013; 177:104-113.
14. Ma Z, Zhang Y, Huet P-M, Lee SS. Differential effects of jaundice and cirrhosis on β-adrenoceptor signaling in three rat models of cirrhotic cardiomyopathy. J Hepatol 1999; 30:485-491.
15. Hu M-L. [41] Measurement of protein thiol groups and glutathione in plasma. Methods in Enzymology 1994; 233:380-385.
16. Beers RF, Sizer IW. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 1952; 195:133-140.
17. Nelson DP, Kiesow LA. Enthalpy of decomposition of hydrogen peroxide by catalase at 25 C (with molar extinction coefficients of H2O2 solutions in the UV). Anal Biochem 1972; 49:474-478.
18. Rukmini M, D’souza B, D’souza V. Superoxide dismutase and catalase activities and their correlation with malondialdehyde in schizophrenic patients. Indian Journal of Clinical Biochemistry 2004; 19:114-118.
19. Barta A, Janega P, Babál P, Murár E, Cebová M, Pechánová O. The effect of curcumin on liver fibrosis in the rat model of microsurgical cholestasis. Food & function 2015; 6:2187-2193.
20. Antoniades C, Bakogiannis C, Tousoulis D, Reilly S, Zhang MH, Paschalis A, et al. Preoperative atorvastatin treatment in CABG patients rapidly improves vein graft redox state by inhibition of Rac1 and NADPH-oxidase activity. Circulation 2010; 122: S66-73.
21. Chang YY, Wu YW, Lee JK, Lin YM, Lin YT, Kao HL, et al. Effects of 12 weeks of atorvastatin therapy on myocardial fibrosis and circulating fibrosis biomarkers in statin-naive patients with hypertension with atherosclerosis. J Investig Med 2016; 64:1194-1199.
22. Sheen JM, Chen YC, Tain YL, Huang LT. Increased circulatory asymmetric dimethylarginine and multiple organ failure: bile duct ligation in the rat as a model. Int J Mol Sci 2014; 15:3989-4006.
23. Dalle-Donne I, Giustarini D, Colombo R, Rossi R, Milzani A. Protein carbonylation in human diseases. Trends Mol Med 2003; 9:169-176.
24. Eskandari-Nasab E, Kharazi-Nejad E, Nakhaee A, Afzali M, Tabatabaei SP, Tirgar-Fakheri K, et al. 50-bp Ins/Del polymorphism of SOD1 is associated with increased risk of cardiovascular disease. Acta Med Iran 2014; 52:591-595.
25. Sallie R, Tredger JM, Williams R. Drugs, and the liver. Part 1: Testing liver function. Biopharm Drug Dispos 1991; 12:251-259.
26. Kluwe J, Mencin A, Schwabe RF. Toll-like receptors, wound healing, and carcinogenesis. J Mol Med (Berl) 2009; 87:125-138.
27. Li Y, Xiong L, Gong J. Lyn kinase enhanced hepatic fibrosis by modulating the activation of hepatic stellate cells. Am J Transl Res 2017; 9:2865-2877.
28. Yang D, Li L, Qian S, Liu L. Evodiamine ameliorates liver fibrosis in rats via TGF-beta1/Smad signaling pathway. J Nat Med 2017.
29. Yang L, Wang Y, Mao H, Fleig S, Omenetti A, Brown KD, et al. Sonic hedgehog is an autocrine viability factor for myofibroblastic hepatic stellate cells. J Hepatol 2008; 48:98-106.
30. Choi SS, Sicklick JK, Ma Q, Yang L, Huang J, Qi Y, et al. Sustained activation of Rac1 in hepatic stellate cells promotes liver injury and fibrosis in mice. Hepatology 2006; 44:1267-1277.
31. Brandes RP, Weissmann N, Schroder K. Nox family NADPH oxidases: Molecular mechanisms of activation. Free Radic Biol Med 2014; 76:208-226.
32. Paik YH, Brenner DA. NADPH oxidase-mediated oxidative stress in hepatic fibrogenesis. Korean J Hepatol 2011; 17:251-257.
33. Li J, Zhu H, Shen E, Wan L, Arnold JM, Peng T. Deficiency of rac1 blocks NADPH oxidase activation, inhibits endoplasmic reticulum stress and reduces myocardial remodeling in a mouse model of type 1 diabetes. Diabetes 2010; 59:2033-2042.