CHOLESTATIC LIVER FIBROSIS IN A RAT MODEL OF BILE DUCT LIGATION: EVALUATING BIOCHEMICAL VERSUS HISTOPATHOLOGICAL CHANGES
Keywords:
Bile duct ligation, Fibrosis, Sirus red, ALT, AST, HydroxyprolineAbstract
Objective: Bile duct ligation (BDL), chronic liver injury model, was extensively used in studying mechanisms of fibrogenesis and antifibrotic agents. Considering the liver regenerative capacity and the diverse results from BDL, the present study aimed to evaluate the biochemical and histopathological changes over 10 weeks following BDL assessing if BDL-induced changes remain in a deterioration state or improve at a certain stage.
Methods: Sham operation and BDL were conducted in Male Wistar rats. Serum AST, ALT, total bilirubin and albumin and hepatic hydroxyproline (HYP), reduced glutathione (GSH) and malondialdehyde (MDA) were measured in sham-operated (n=3) and BDL-rats (n=6) at 0, 1, 2, 4, 6, 8 and 10 weeks following operation. Liver tissue was also processed for histopathological analysis (H&E and Sirus red staining).
Results: Progressive liver injury (H&E) and collagen deposition (Sirus red and HYP) in BDL-rats were observed starting from the first week post-operation and reached their maximum with early signs of cirrhosis on the 10th week of BDL. Severe and sustained cholestatic injury appeared in 2 weeks (increased ALT, AST, bilirubin along with decreased albumin (P<0.001) compared to sham-operated rats). AST peaked on first week, however, bilirubin, ALT and MDA peaked on the 4th week (P<0.001) then gradually decreased compared to their peaks.
Conclusion: The relative improvement in liver function/cholestasis following their peaks in BDL model despite progression of fibrosis and hepatic injury require investigators using this model to consider not only biochemical, but also histopathological findings to guarantee an accurate interpretation of their results.
Â
Downloads
References
Liu Y, Meyer C, Xu C, Weng H, Hellerbrand C, ten Dijke P, Dooley S. Animal models of chronic liver diseases. Am J Physiol Gastrointest Liver Physiol 2013;304:G449-68.
Bataller R, Brenner DA. Liver fibrosis. J Clin Invest 2005;115:209-18.
Lai HS, Wu YM, Lai SL, Lin WH. Lipocalin-2 gene expression during liver regeneration after partial hepatectomy in rats. Int J Surg 2013;11:314-8.
Best J, Dolle L, Manka P, Coombes J, van Grunsven LA, Syn WK. Role of liver progenitors in acute liver injury. Front Physiol 2013;4:258.
Ramadan A, Soliman G, Mahmoud SS, Nofal SM, Abdel-rahman RF. Hepatoprotective and hepatotheraputic effects of propolis against d-galactosamine/lipopolysaccharide-induced liver damage in rats. IJPPS 2015;7:372-8.
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. Fibrog Tissue Repair 2013;6:19.
Gujral JS, Liu J, Farhood A, Hinson JA, Jaeschke H. Functional importance of icam-1 in the mechanism of neutrophil-induced liver injury in bile duct-ligated mice. Am J Physiol: Gastrointest Liver Physiol 2004;286:G499-507.
Yang M, Ramachandran A, Yan HM, Woolbright BL, Copple BL, Fickert P, et al. Osteopontin is an initial mediator of inflammation and liver injury during obstructive cholestasis after bile duct ligation in mice. Toxicol Lett 2014;224:186-95.
Popov Y, Schuppan D. Targeting liver fibrosis: Strategies for development and validation of antifibrotic therapies. Hepatology 2009;50:1294-306.
Heinrich S, Georgiev P, Weber A, Vergopoulos A, Graf R, Clavien PA. Partial bile duct ligation in mice: A novel model of acute cholestasis. Surgery 2010;149:445-51.
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;50:3505-13.
Yu Y, Zhang F, Lü L, Fan Y, Li G, Wang X. Construction of hepatocyte growth factor gene recombinant adenovirus vector and its expression in rat bone marrow mesenchymal stem cells. J Nanjing Med Univ 2008;22(2):81-6.
Tarcin O, Avsar K, Demirturk L, Gultepe M, Oktar BK, Ozdogan OC, et al. In vivo inefficiency of pentoxifylline and interferon-alpha on hepatic fibrosis in biliary-obstructed rats: Assessment by tissue collagen content and prolidase activity. J Gastroenterol Hepatol 2003;18:437-44.
Panakanti R, Pratap A, Yang N, Jackson JS, Mahato RI. Triplex forming oligonucleotides against type alpha1(i) collagen attenuates liver fibrosis induced by bile duct ligation. Biochem Pharmacol 2010;80:1718-26.
Georgiev P, Jochum W, Heinrich S, Jang JH, Nocito A, Dahm F, et al. Characterization of time-related changes after experimental bile duct ligation. Br J Surg 2008;95:646-56.
El-Swefy S, Hassanen SI. Improvement of hepatic fibrosis by leukotriene inhibition in cholestatic rats. Ann Hepatol 2009;8:41-9.
Fursule RA, Patil SD. Hepatoprotective and antioxidant activity of phaseolus trilobus, ait on bile duct ligation induced liver fibrosis in rats. J Ethnopharmacol 2010;129:416-9.
Kim HG, Han JM, Lee HW, Lee JS, Son SW, Choi MK, et al. Cgx, a multiple herbal drug, improves cholestatic liver fibrosis in a bile duct ligation-induced rat model. J Ethnopharmacol 2013;145:653-62.
Liu Y, Binz J, Numerick MJ, Dennis S, Luo G, Desai B, et al. Hepatoprotection by the farnesoid x receptor agonist gw4064 in rat models of intra-and extrahepatic cholestasis. J Clin Invest 2003;112:1678-87.
Fujita M, Shannon JM, Morikawa O, Gauldie J, Hara N, Mason RJ. Overexpression of tumor necrosis factor-alpha diminishes pulmonary fibrosis induced by bleomycin or transforming growth factor-beta. Am J Respir Cell Mol Biol 2003;29:669-76.
Fiel MI. Histologic scoring systems for chronic liver disease; in Chopra S (ed: Up To Date, Wolters Kluwer Health; 2007, 2015.
Lee S, Kim S, Le HD, Meisel J, Strijbosch RA, Nose V, Puder M. Reduction of hepatocellular injury after common bile duct ligation using omega-3 fatty acids. J Pediatr Surg 2008;43:2010-5.
Beaussier M, Wendum D, Schiffer E, Dumont S, Rey C, Lienhart A, et al. Prominent contribution of portal mesenchymal cells to liver fibrosis in ischemic and obstructive cholestatic injuries. Lab Invest 2007;87:292-303.
Marques TG, Chaib E, da Fonseca JH, Lourenco AC, Silva FD, Ribeiro MA, et al. Review of experimental models for inducing hepatic cirrhosis by bile duct ligation and carbon tetrachloride injection. Acta Cirurgica Brasileira 2012;27:589-94.
Dhanda S, Kaur S, Sandhir R. Preventive effect of n-acetyl-l-cysteine on oxidative stress and cognitive impairment in hepatic encephalopathy following bile duct ligation. Free Radic Biol Med 2013;56:204-15.
Copple BL, Jaeschke H, Klaassen CD. Oxidative stress and the pathogenesis of cholestasis. Semin Liver Dis 2010;30:195-204.
Vinoth Prabhu V, Chidambaranathan N, Nalini G, Venkataraman S, Jayaprakash S, Nagarajan M. Evaluation of anti-fibrotic effect of lagerstroemia speciosa (l) pers. On carbon tetrachloride induced liver fibrosis. Curr Pharm Res 2010;1(1):7-12.
Dai LJ, Li HY, Guan LX, Ritchie G, Zhou JX. The therapeutic potential of bone marrow-derived mesenchymal stem cells on hepatic cirrhosis. Stem Cell Res 2009;2:16-25.
Faropoulos K, Chroni E, Assimakopoulos SF, Mavrakis A, Stamatopoulou V, Toumpeki C, et al. Altered occludin expression in brain capillaries induced by obstructive jaundice in rats. Brain Res 2010;1325:121-7.
Tarcin O, Basaranoglu M, Tahan V, Tahan G, Sucullu I, Yilmaz N, et al. Time course of collagen peak in bile duct-ligated rats. BMC Gastroenterol 2011;11:45.
Trauner M, Boyer JL. Bile salt transporters: Molecular characterization, function, and regulation. Physiol Rev 2003;83:633-71.
Eipel C, Menschikow E, Sigal M, Kuhla A, Abshagen K, Vollmar B. Hepatoprotection in bile duct ligated mice mediated by darbepoetin-alpha is not caused by changes in hepatobiliary transporter expression. Int J Clin Exp Pathol 2013;6:80-90.
Roma MG, Crocenzi FA, Sanchez Pozzi EA. Hepatocellular transport in acquired cholestasis: New insights into functional, regulatory and therapeutic aspects. Clin Sci (Lond) 2008;114:567-88.
Froh M, Zhong Z, Walbrun P, Lehnert M, Netter S, Wiest R, et al. Dietary glycine blunts liver injury after bile duct ligation in rats. World J Gastroenterol 2008;14:5996-6003.
Krahenbuhl S, Talos C, Lauterburg BH, Reichen J. Reduced antioxidative capacity in liver mitochondria from bile duct ligated rats. Hepatology 1995;22:607-12.
Ohta Y, Kongo M, Sasaki E, Nishida K, Ishiguro I. Therapeutic effect of melatonin on carbon tetrachloride-induced acute liver injury in rats. J Pineal Res 2000;28:119-26.
Suzuki A, Angulo P, Lymp J, Li D, Satomura S, Lindor K. Hyaluronic acid, an accurate serum marker for severe hepatic fibrosis in patients with non-alcoholic fatty liver disease. Liver Int 2005;25:779-86.
Henriksen JH, Siemssen O, Krintel JJ, Malchow-Moller A, Bendtsen F, Ring-Larsen H. Dynamics of albumin in plasma and ascitic fluid in patients with cirrhosis. J Hepatol 2001;34:53-60.
Bernardi M, Maggioli C, Zaccherini G. Human albumin in the management of complications of liver cirrhosis. Crit Care 2012;16:211.
Rivera-Huizar S, Rincon-Sanchez AR, Covarrubias-Pinedo A, Islas-Carbajal MC, Gabriel-Ortiz G, Pedraza-Chaverri J, et al. Renal dysfunction as a consequence of acute liver damage by bile duct ligation in cirrhotic rats. Exp Toxicol Pathol 2006;58:185-95.
Sheen JM, Huang LT, Hsieh CS, Chen CC, Wang JY, Tain YL. Bile duct ligation in developing rats: Temporal progression of liver, kidney, and brain damage. J Pediatr Surg 2010;45:1650-8.
Aktas C, Kanter M, Erboga M, Mete R, Oran M. Melatonin attenuates oxidative stress, liver damage and hepatocyte apoptosis after bile-duct ligation in rats. Toxicol Ind Health 2012;30:835-44.
Chen CY, Wang BT, Wu ZC, Yu WT, Lin PJ, Tsai WL, et al. Glycine ameliorates liver injury and vitamin d deficiency induced by bile duct ligation. Clin Chim Acta 2013;420:150-4.
Muchova L, Vanova K, Zelenka J, Lenicek M, Petr T, Vejrazka M, et al. Bile acids decrease intracellular bilirubin levels in the cholestatic liver: Implications for bile acid-mediated oxidative stress. J Cell Mol Med 2011;15:1156-65.
Wang G, Shen H, Rajaraman G, Roberts MS, Gong Y, Jiang P, et al. Expression and antioxidant function of liver fatty acid binding protein in normal and bile-duct ligated rats. Eur J Pharmacol 2007;560:61-8.
Shetty SR, Babu SG, Kumari S, Rao V, Vijay R, Karikal A. Malondialdehyde levels in oral sub mucous fibrosis: A clinicopathological and biochemical study. N Am J Med Sci 2012;4:125-8.
Maher JJ, Tzagarakis C, Gimenez A. Malondialdehyde stimulates collagen production by hepatic lipocytes only upon activation in primary culture. Alcohol Alcohol 1994;29:605-10.