Hepatoprotective and antifibrotic effects of trans-chalcone against bile duct ligation-induced liver fibrosis in rats

Document Type : Original Article


1 Department of Pharmacology and Toxicology, Faculty of Pharmacy and Pharmaceutical Sciences, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

2 Department of Physiology, Faculty of Medicine, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran

3 Department of Pathology, Faculty of Specialized Veterinary, Science and Research Branch, Islamic Azad University, Tehran, Iran


Objective(s): Several lines of research have shown that hepatic fibrosis is one of the leading causes of death worldwide. Trans-chalcone is a flavonoid precursor with anti-oxidant and anti-inflammatory effects. The present study was conducted to examine the antifibrotic properties of trans-chalcone on bile duct ligation (BDL)-induced liver cholestasis in rats. 
Materials and Methods: Following the BDL operation, trans-chalcone at doses of 12, 24, and 50 mg/kg was administered orally once a day for 45 consecutive days. Serum levels of liver indices, including alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total and direct bilirubin, and lipid profile in addition to blood urea nitrogen (BUN) and creatinine, were measured. Additionally, catalase (CAT) and superoxide dismutase (SOD) activities were assessed in liver homogenates. Histopathological evaluations were performed using Masson trichrome (MT) and hematoxylin and eosin (H&E) staining. 
Results: The elevated levels of liver enzymes, total and direct bilirubin, BUN, creatinine, cholesterol, triglyceride, and low-density lipoprotein (LDL) induced by BDL were significantly reduced following trans-chalcone administration; while serum level of high-density lipoprotein (HDL) increased. Besides, treatment with trans-chalcone elevated the activities of CAT and SOD in the liver tissues of the animals with BDL surgery. According to MT and H&E staining, BDL-induced histopathological changes, including infiltration of inflammatory cells, hepatocyte necrosis, ductal hyperplasia, and collagen deposition were ameliorated using trans-chalcone administration. 
Conclusion: It can be concluded from the present study that trans-chalcone, possibly by its anti-oxidant and anti-inflammatory properties, may exert hepatoprotective and antifibrotic effects in BDL-induced liver fibrosis. v


Main Subjects

 1. Ingawale DK, Mandlik SK, Naik SR. Models of hepatotoxicity and the underlying cellular, biochemical and immunological mechanism (s): a critical discussion. Environ Toxicol Pharmacol 2014; 37:118-133.
2. Yin C, Evason KJ, Asahina K, Stainier DYR. Hepatic stellate cells in liver development, regeneration, and cancer. J Clin Invest 2013; 123:1902-1910.
3. Chen WY, Chen CJ, Liao JW, Mao FC, et al. Chromium attenuates hepatic damage in a rat model of chronic cholestasis. Life Sci 2009; 84:606-614.
4. Cruz A, Francisco J, Padillo FJ, Granados J, Tunez I, Carmen Munoz M, et al. Effect of melatonin on cholestatic oxidative stress under constant light exposure. Cell Biochem Funct 2003; 21:377-380.
5. Yang JH, Kim SC, Kim KM, Jang CH, Cho SS, Kim SJ, et al. Isorhamnetin attenuates liver fibrosis by inhibiting TGF-β/smad signaling and relieving oxidative stress. Eur J Pharmacol 2016; 783:92-102.
6. Han JM, Kim HG, Choi MK, Lee JS, Park HJ, Wang JH, et al. Aqueous extract of artemisia iwayomogi kitamura attenuates cholestatic liver fibrosis in a rat model of bile duct ligation. Food Chem Toxicol 2012; 50:3505-3513.
7. Karkhaneh L, Yaghmaei P, Parivar K, Sadeghizadeh M, Ebrahim-Habibi A. Effect of trans-chalcone on atheroma plaque formation, liver fibrosis and adiponectin gene expression in cholesterol-fed NMRI mice. Pharmacol Rep 2016; 68:720-727.
8. Singh H, Siduh S, Chopra K, Khan MU. Hepatoprotective effect of trans-chalcone on experimentally induced hepatic injury in rats: inhibition of hepatic inflammation and fibrosis. Can J Physiol Pharmacol 2016; 94:879-887.
9. Jalalvand F, Amoli MM, Yaghmaei P, Kimiagar M, Ebrahim-Habibi A. Acarbose versus trans-chalcone: comparing the effect of two glycosidase inhibitors on obese mice. Arch Endocrinol Metab 2015; 59:202-209.
10. Karimi-Sales E, Ebrahimi-Kalan A, Alipour MR. Preventive effect of trans-chalcone on non-alcoholic steatohepatitis: Improvement of hepatic lipid metabolism. Biomed Pharmacother 2019; 109:1306-1312.
11. Karimi-Sales E, Jadidi S, Ebrahimi-Kalan A, Alipour MR. Trans-chalcone prevents insulin resistance and hepatic inflammation and also promotes hepatic cholesterol efflux in high-fat diet-fed rats: modulation of miR-34a-, miR-451-, and miR-33a-related pathways. Food  Funct 2018; 9:4292-4298.
12. Singh H, Sidhu S, Khan M. Free radical scavenging property of β-aescin and trans-chalcone: in vitro study. Eur J Pharm Med Res 2016; 3:309-312.
13. Staurengo-Ferrari L, Ruiz-Miyazawa KW, Pinho-Ribeiro FA, Fattori V, Zaninelli Th, Badaro-Garcia S, et al. Trans-chalcone attenuates pain and inflammation in experimental acute gout arthritis in mice. Front Pharmacol 2018; 9:1123-1138.
14. Sikander M, Malik S, Yadav D, Biswas S, Katare DP, Jain SK. Cytoprotective activity of a trans-chalcone against hydrogen peroxide induced toxicity in hepatocellular carcinoma (HepG2) cells. Asian Pac J Cancer Prev 2011; 12:2513-2516.
15. Uchinami H, Seki E, Brenner DA,  D’Armiento J. Loss of MMP 13 attenuates murine hepatic injury and fibrosis during cholestasis. Hepatology 2006; 44:420-429.
16. Gross Jr J B, Reichen J, Zeltner TB, Zimmermann A. The evolution of changes in quantitative liver function tests in a rat model of biliary cirrhosis: correlation with morphometric measurement of hepatocyte mass. Hepatology 198; 7:457-463.
17. Rifai N, Tietz Textbook of Clinical Chemistry and Molecular Diagnostics-e-Book. 2017: Elsevier Health Sciences.
18. Moss DW. Clinical enzymology. Nature 1971; 233:505-505.
19. Aebi, H. [13] Catalase in vitro. Meth Enzymol 1984; 105:121-126.
20. Shan BS, Mogi M, Iwanami J, Bai HY, No HK, Higaki A, et al. Attenuation of stroke damage by angiotensin II type 2 receptor stimulation via peroxisome proliferator-activated receptor-gamma activation. Hypertens Res 2018; 41:839-848.
21. Sant’Anna LB, Cargnoni A, Ressel L, Vanosi G, Parolini O. Amniotic membrane application reduces liver fibrosis in a bile duct ligation rat model. Cell Transplant 2011; 20:441-453.
22. French SW, Miyamoto K, Ohta Y, Geoffrion Y. Pathogenesis of experimental alcoholic liver disease in the rat. Meth Achiev Exp Pathol 1988; 13:181-207.
23. Perez MJ, Briz O. Bile-acid-induced cell injury and protection. World J Gastroenterol 2009; 15:1677-1689.
24. Sokol RJ, Straka MS, Dahl R, Devereaux MW, Yerushalmi B, Gumpricht E, et al. Role of oxidant stress in the permeability transition induced in rat hepatic mitochondria by hydrophobic bile acids. Pediatr Res 2001; 49:519-531.
25. Ale-Ebrahim M, Eidi A, Mortazavi P, Tavangar SM, Tehrani DM. Hepatoprotective and antifibrotic effects of sodium molybdate in a rat model of bile duct ligation. J Trace Elem Med Biol 2015; 29:242-248.
26. Ramaiah SK. A toxicologist guide to the diagnostic interpretation of hepatic biochemical parameters. Food Chem Toxicol 2007; 45:1551-1557.
27. Karimi-Sales E, Jadidi S, Ghaffari-Nasab A, Salimi M, Alipour MR. Effect of trans-chalcone on hepatic IL-8 through the regulation of miR-451 in male rats. Endocr Regul 2018; 52:1-5.
28. Ale-Ebrahim M, Rahmani R, Faryabi K, Mohammadifar N, Mortazavi P, Karkhaneh L. Atheroprotective and hepatoprotective effects of trans-chalcone through modification of eNOS/AMPK/KLF-2 pathway and regulation of COX-2, Ang-II, and PDGF mRNA expression in NMRI mice fed HCD. Mol Biol Rep 2022; 49:3433-3443.
29. Karimi-Sales E, Sajadi J, Ebrahimi-Kalan A, Alipour MR. Protective role of trans-chalcone against the progression from simple steatosis to non-alcoholic steatohepatitis: regulation of miR-122, 21, 34a, and 451. Adv Pharm Bull 2022; 12:200-205.
30. Trauner M, Meier PJ, Boyer LJ. Molecular pathogenesis of cholestasis. N Engl J Med 1998; 339:1217-1227.
31. Orellana M, Rodrigo R, Thielemann L, Guajardo V. Bile duct ligation and oxidative stress in the rat: effects in liver and kidney. Comp Biochem Physiol Toxicol Pharmacol 2000; 126:105-111.
32. Sokol RJ, Devereaux M, Khandwala R, O’Brien K. Evidence for involvement of oxygen free radicals in bile acid toxicity to isolated rat hepatocytes. Hepatology 1993; 17:869-881.
33. Unsal V, Deveci K, Ozmen ZC, Tumer MK. Research on the effects of L-carnitine and trans-chalcone on endoplasmic reticulum stress and oxidative stress in high-fructose corn syrup-fed rats. Nutr Food Sci 2020; 51:1-17.
34. Martinez RM, Pinho-Ribeiro FA, Vale DL, Steffen VS, Vicentini FTMC, Vignoli JA, et al. Trans-chalcone added in topical formulation inhibits skin inflammation and oxidative stress in a model of ultraviolet B radiation skin damage in hairless mice. J Photochem Photobiol B 2017;  171:139-146.
35. Ji H, Jiang JY, Xu Z, Kroeger EA, Lee SS, Liu H, et al. Change in lipid profile and impairment of endothelium-dependent relaxation of blood vessels in rats after bile duct ligation. Life Sci 2003; 73:1253-1263.
36. Longo M, Crosignani A, Podda M. Hyperlipidemia in chronic cholestatic liver disease. Curr Treat Options Gastroenterol 2001; 4:111-114.
37. Delgado-Villa MJ, Ojeda ML, Rubio JM, Murillo ML, Sanchez OC. Beneficial role of dietary folic acid on cholesterol and bile acid metabolism in ethanol-fed rats. J Stud Alcohol Drugs 2009; 70:615-622.
38. Nuño‐Lámbarri N, Barbero-Becerra VJ, Uribe M, Chávez-Tapia NC. Elevated cholesterol levels have a poor prognosis in a cholestasis scenario. J Biochem Mol Toxicol 2017; 31:1-6.
39. Claudel T, Sturm E, Duez H, Torra IP, Sirvent A, Kosykh V, et al. Bile acid-activated nuclear receptor FXR suppresses apolipoprotein AI transcription via a negative FXR response element. J Clin Invest 2002; 109:961-971.
40. Najafian M, Ebrahim-Habibi A, Yaghmaei P, Parivar K, Larijani B. Core structure of flavonoids precursor as an antihyperglycemic and antihyperlipidemic agent: an in vivo study in rats. Acta Biochim Pol 2010; 57:553-560.
41. Laliotis GP, Bizelis I, Rogdakis E. Comparative approach of the de novo fatty acid synthesis (lipogenesis) between ruminant and non ruminant mammalian species: from biochemical level to the main regulatory lipogenic genes. Curr Genomics 2010; 11:168-183.
42. Kemper JK, Choi SE, Kim DH. Sirtuin 1 deacetylase: a key regulator of hepatic lipid metabolism. Vitam Horm 2013; 91:385-404.
43. Bitencourt TA, Komoto TT, Massaroto BG, Saraiva Miranda CE, Beleboni RO, Marins M, et al. Trans-chalcone and quercetin down-regulate fatty acid synthase gene expression and reduce ergosterol content in the human pathogenic dermatophyte Trichophyton rubrum. BMC Complement Altern Med 2013; 13:1-6.
44. Casini A, Pinzani M, Milani S, Grappone C, Galli G, Jezequel AM, et al. Regulation of extracellular matrix synthesis by transforming growth factor β1 in human fat-storing cells. Gastroenterology 1993; 105:245-253.
45. Ramadori G, Knittel T, Odenthal M, Schwögler S, Neubauer K, Meyer zum Büschenfelde KH. Synthesis of cellular fibronectin by rat liver fat-storing (Ito) cells: regulation by cytokines. Gastroenterology 1992; 103:1313-1321.
46. Wei Y, Clark SE, Thyfault JP, Uptergrove GME, Li W, Whaley-Connell AT, et al. Oxidative stress-mediated mitochondrial dysfunction contributes to angiotensin II-induced nonalcoholic fatty liver disease in transgenic Ren2 rats. Am J Pathol 2009; 174:1329-1337.
47. Ying HZ, Chen Q, Zhang WY, Zhang HH, Ma Y, Zhang SZ, et al. PDGF signaling pathway in hepatic fibrosis pathogenesis and therapeutics. Mol Med Rep 2017; 16:7879-7889.
48. Karimi-Sales E, Jeddi S, Alipour MR. Trans-chalcone inhibits transforming growth factor-β1 and connective tissue growth factor-dependent collagen expression in the heart of high-fat diet-fed rats. Arch Physiol Biochem 2020; 128:1221-1224.
49. Karimi-Sales E, Jeddi S, Alipour MR. Protective effect of trans-chalcone against high-fat diet-induced pulmonary inflammation is associated with changes in miR-146a and pro-inflammatory cytokines expression in male rats. Inflammation 2019; 42:2048-2055.
50. Rivera-Huizar S, Rincón-Sánchez AR, Covarrubias-Pinedo A, Islas-Carbajal MC, Gabriel-Ortíz G, Pedraza-Chaverrí J, et al. Renal dysfunction as a consequence of acute liver damage by bile duct ligation in cirrhotic rats. Exp Toxicol Pathol 2006; 58:185-195.
51. Yan CG, Zhu DF, Wang F. Study on the expressions and roles of renal heat shock protein 72 and toll-like receptor 4 in hepatorenal syndrome in rat. Zhongguo wei Zhong Bing ji jiu yi xue 2007; 19:731-734.
52. Alipour MR, Jeddi S, Karimi‐Sales E. Trans‐chalcone inhibits high‐fat diet‐induced disturbances in FXR/SREBP‐1c/FAS and FXR/Smad‐3 pathways in the kidney of rats. J Food Biochem 2020; 44:13476-13483.