(1. School of Pharmacy, Jiangsu University, Zhenjiang Jiangsu 212013； 2. School of Medicine, Jiangsu University, Zhenjiang Jiangsu 212013； 3. School of Pharmacy, Soochow University, Suzhou Jiangsu 215123； 4. School of Life Sciences, Nanjing University, Nanjing Jiangsu 210093, China)
Abstract：Objective: To investigate the effect and mechanism of asiatic acid(AA) on experimental cholestasisinduced liver injury. Methods: In vivo, a total of 32 C57 mice were divided into sham group, bile duct ligation(BDL) model group, AA 15 mg/kg group and AA 30 mg/kg group. Mice were anesthetized by an intraperitoneal (i.p) injection of pentobarbital (60 mg/kg). After anaesthetized, the common bile duct was doubleligated by suture. The sham group and model group were intragastric administration by 0.5% CMCNa solution, the other two groups were intragastric administration by AA (dissolved in 0.5% CMCNa). All groups were intragastric administration with CMCNa or AA per day for 5 days. The morphology of liver were observed，and also the content of glutamicoxalacetic transaminase(AST) and glutamicpyruvic transaminase(ALT) biochemically determined. Liver tissue sections were used for H＆E staining, Masson staining and Tunel staining.PCR assay tested the expression levels of inflammatory factors including prostaglandinendoperoxide synthase(PTGS2), IL6,TNFα, Chemokine ligand 3 (CCL3). The expression levels of apoptosisrelated proteins (Caspase3, cleavedcaspase3, Bax, Bcl2) were determined using Western blotting. In vitro, the cell viability ratio and apoptosis of HL7702 treated with sodium glycochenodeoxycholate(GCDC) detected by MTT and flow cytometry assay, respectively. Results: In vivo, after treated with AA, the liver index and spleen index of mice were significantly reduced, compared with model group. AA decreased the content of AST and ALT, and reduced the hepatocyte apoptosis showed by Tunel staining. And also, AA decreased the level of proinflammation genes and regulated the expression of apoptosisrelated proteins.AA can inhibit GCDCinduced HL7702 cells apoptosis in a concentrationdependent manner. Conclusion: Asiatic acid can alleviate liver injury by inhibiting hepatocyte apoptosis.
[1\]Schuppan D,Kim YO.Evolving therapies for liver fibrosis\[J\].J Clin Invest,2013, 123(5): 1887-1901.\[2\]Mederacke I, Hsu CC, Troeger JS, et al. Fate tracing reveals hepatic stellate cells as dominant contributors to liver fibrosis independent of its aetiology\[J\]. Nat Commun, 2013, 4(7):657-678.\[3\]Shio S, Yazumi S, Ogawa K, et al. Biliary complications in donors for living donor liver transplantation\[J\]. Am J Gastroenterol, 2008, 103(6):1393-1398.\[4\]Jüngst C, Lammert F. Cholestatic liver disease\[J\]. Dig Dis, 2013,31(1): 152-154.\[5\]Granato A,Gores G,Vilei MT, et al. Bilirubin inhibits bile acid induced apoptosis in rat hepatocytes\[J\]. Gut, 2003, 52(12):1774-1778.\[6\]Ramachandran V,Saravanan R. Efficacy of asiatic acid，a pentacyclic triterpene on attenuating the key enzymes activities of carbohydrate metabolism in streptozotocininduced diabetic rats\[J\]. Phytomedicine, 2013, 20(3/4):230-236.\[7\]Ma K,Zhang Y,Zhu D,et al.Protective effects of asiatic acid against Dgalactosamine/lipopolysaccharideinduced hepatotoxicity in hepatocytes and kupffer cells cocultured system via redoxregulated leukotriene C4 synthase expression pathway\[J\]. Eur J Pharmacol,2009,603(1): 98-107.\[8\]Huang SS，Chiu CS，Chen HJ，et al. Antinociceptive activities and the mechanisms of antiinflammation of asiatic acid in mice\[J\]. Evid Based Complement Alternat Med, 2011, 2011: 895857.\[9\]Singh RP, Agarwal R. Inducible nitric oxide synthasevascular endothelial growth factor axis: a potential target to inhibit tumor angiogenesis by dietary agents\[J\]. Curr Cancer Drug Tar, 2007, 7( 5) : 475-483.\[10\]Liuv TZ, Lee KT, Chern CL, et al. Free radicaltriggered hepatic injury of experimental obstructive jaundice of rats involves overproduction of proinflammatory cytokines and enhanced activation of nuclear factor κB\[J\]. Ann Clin Lab Sci,2001, 31(4):383-390.\[11\]Ramm GA, Nair VG, Bridle KR, et al. Contribution of hepatic parenchymal and nonparenchymal cells to hepatic fibrogenesis in biliary atresia\[J\]. Am J Pathol, 1998, 153(2):527-535.\[12\]Jiang JX, Trk NJ. Liver injury and the activation of the hepatic myofibroblasts\[J\]. Curr Pathobiol Rep, 2013, 1(3):215-223.\[13\]Miyoshi H, Rust C, Roberts PJ, et al. Hepatocyte apoptosis after bile duct ligation in the mouse involves Fas\[J\]. Gastroenterology, 1999, 117(3):669-677.
2017, Vol. 27 
