盐酸小檗碱对肝纤维化大鼠肠道黏膜屏障的保护作用及机制

Protective effect of berberine hydrochloride on intestinal mucosal barrier in rats with hepatic fibrosis

目的建立肝纤维化大鼠模型,观察盐酸小槃碱对肝纤维化大鼠肠道黏膜屏障的保护作用,并探讨其机制。方法将大鼠随机分为正常对照组(CON组)、模型组(MOD组)、乳果糖口服液组(LOL组)和盐酸小槃碱组(BH组),除CON组外,其他各组建立肝纤维化模型,LOL组按100 m/kg的剂量灌胃乳果糖口服溶液,BH组按150 mg/kg的剂量灌胃盐酸小槃碱药液,CON组和MOD组按10 ml/kg的剂量灌胃生理盐水。各组连续灌胃12周。灌胃结束后,计算肝脏指数和脾脏指数;检测各组大鼠血清中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、透明质酸钠(HA)、层粘连蛋白(LN)、IV型胶原蛋白(IA-C)及血清内毒素的含量.用HE染色观察肝脏和回肠病理变化;用实时荧光定量聚合酶链式反应(RT-PCR)和蛋白质印迹法(Western blot)检测回肠中闭合蛋白(Claudins)、闭锁连接蛋白-1(ZO-1)的mRNA和蛋白表达水平。结果乳果糖口服液和盐酸小槃碱可不同程度减轻大鼠肝脏指数和脾脏指数(P<0.01,P<0.05).乳果糖口服液和盐酸小槃碱可减少血清中ALT、AST、ALP的含量(P<0.01,P<0.05)。乳果糖口服液和盐酸小槃碱可减少大鼠血清中HA、LN'IVC及内毒素的含量(2<0,01,P<0.05).乳果糖口服液和盐酸小槃碱改善了大鼠肝细胞、肝小叶、肝索的结构,并能减少炎性细胞浸润.乳果糖口服液和盐酸小槃碱可减少回肠肠黏膜充血、水肿及炎性细胞浸润.乳果糖口服液和盐酸小槃碱均提高大鼠回肠中Cmudins、ZOO mRNA和蛋白表达水平(P<0.01,P<0.05).结论盐酸小槃碱对肝纤维化大鼠肠道黏膜屏障具有保护作用,其机制可能与其调节回肠中肠道屏障保护蛋白Cmudins、ZOO的表达有关。

Objective To investigate the protective effects of berberine hydrochloride on intestinal mucosal barrier in rats with liver fibrosis and explore its mechanism.Methods Rats were randomly divided into control group(CON group),model group(MOD group),lactulose oral liquid group(L0L group)and berberine hydrochloride group(BH group).In addition to CON group,other groups established liver fibrosis models.In the LOL group,the lactulose oral liquid was administered at a dose of 100 mg/kg,the BH group was administered with berberine liquid at a dose of 150 mg/kg,and the CON group and the MOD group were orally administered with normal saline at a dose of 10 ml/kg.Each group was continuously administered for 12 weeks.After gastric administration,liver index and spleen index were calculated.The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),sodium hyaluronate(HA),laminin(LN),type IV collagen(IV-C)and endotoxin in.serum of rats in each group were detected.Liver and ileal pathological changes were observed by HE staining.Real time-PCR and W esterm blot were used to detect the expression of mRNA and protein of occlusion protein(Occludin)and atresia conmexin-1(Z0-1)in ileum.Results Lactulose oral liquid and berberine could alleviate the liver index and spleen index of rats in different degrees(P<0.01,P<0.05).Lactulose oral liquid and berberine could reduce the levels of ALT,AST and ALP in serum(P<0.01,P<0.05).Lactulose oral liquid and berberine could reduce the content of HA,LN,IV-C and endotoxin in rat serum(P<0.01,P<0.05).Lactulose oral liquid and berberine improve the structure of rat hepatocytes,hepatic lobules and hepatic cords,and could reduce inflammatory cell infil-tration.Lactulose oral liquid and berberine could reduce collateral intestinal mucosal congestion,edema and inflam-matory cell infiltration.Both lactulose oral liquid and berberine increased the expression of Claudins and Z0-1 mR-NA and protein in rat ileum(P<0.01,P<0.05).Conclusion Bbrberine hycrochloride has protective ffeets on intestinal mucosal barrier in rats with liver fibrosis,and its mechanism may be related to the regulation of the expres-sion of intestinal barrier proteins Claudins and Z0-1.