桃红芪术软肝煎对肝纤维化大鼠病理及TGF－β/Smad信号通路的影响

摘要

目的:观察桃红芪术软肝煎抗大鼠肝纤维化的作用及对TGF－β/Smad信号通路的影响.方法:选取105只Wistar大鼠,随机分为空白组、模型组、对照药物组(秋水仙碱组、扶正化瘀胶囊组)、桃红芪术软肝煎组(桃红芪术低剂量组、中剂量组、高剂量组).除空白组外,分3、6、9周3个时间点采用50%CCl41 mL／kg橄榄油溶液腹腔注射进行肝纤维化模型制作,同时进行HE染色和Masson染色观察肝纤维化的病理表现;并采用免疫组织化学法检测肝脏组织的E－Cadherin、Vim－entin、转化生长因子(TGF)－β1、Smad2 蛋白表达;RT－PCR方法检测肝脏组织的E－Cadherin、Vimentin、TGF－β1、Smad2 的mRNA表达.结果:肝纤维化病理结果显示,随着造模、给药时间的延长,各给药组均能明显阻断模型大鼠肝组织纤维化进展;与模型组比较,第9周时桃红芪术高剂量组作用最明显.免疫组织化学结果显示,与模型组比较,用药各组从第3周至第9周,E－cadherin表达均较模型组表达明显增加(P＜0.05),用药各组间比较,差异无统计学意义( P＞0.05);与模型组比较,用药各组TGF－β1表达明显减弱,差异有统计学意义(P＜0.05),用药各组组间比较,差异无统计学意义( P＞0.05). RT－PCR结果显示,与空白组比较,模型组Smad2 mRNA表达量明显升高,差异有统计学意义(P＜0.05),桃红芪术低剂量组Vimentin mRNA、Smad2 mRNA表达量明显下降,差异有统计学意义(P＜0.05),桃红芪术软肝煎中剂量组Smad2 mRNA表达下降,差异有统计学意义(P＜0.05);扶正化瘀胶囊组TGF－β1mRNA、Vimentin mRNA表达量均较模型组下降,差异有统计学意义(P＜0.05).结论:桃红芪术软肝煎可持续通过TGF－β／Smad信号通路调节上皮间质转化,治疗肝纤维化.%Objective:To observe effects of Taohong Qizhu Ruangan Decoction on treating liver fibrosis of rats , and effects on TGF－β/Smad signal pathway.Methods:A total of 105 Wistar rats were randomly divided into blank group , model group, control group (colchicine and Fuzheng Huayu Capsule group ), Taohong Qizhu Ruangan Decoction group (low dose group, middle dose group, high dose group).Except blank group, the liver fibrosis model was established by the intraperitoneal injection of 50%CCl4 1 mL/kg of olive oil solution at 3, 6, 9 week.At the same time, HE staining and Masson staining were used to observe the patho－logical expressions of liver fibrosis.The expressions of E－Cadherin, Vimentin, TGF－β1and Smad2 in liver tissues were detected by immunohistochemistry.The mRNA expressions of E－Cadherin, Vimentin, TGF－β1and Smad2 in liver tissues were detected by RT－PCR.Results:The pathological results of liver fibrosis showed that with the prolongation of modeling and administration time , each drug－administered group could significantly block the progression of hepatic fibrosis in model rats .And compared with model group , effect of high dose Taohong Qizhu Ruangan Decoction group was most obvious .Immunohistochemical:From 3 week to 9 week, com－ pared with model group, E－cadherin expression was significantly increased than other groups (P＜0.05), and there were no differ－ences between groups (P＞0.05); compared with model group, TGF－βexpression decreased significantly in other drug groups (P＜0.05), and there were no differences between groups (P＞0.05).RT－PCR:Compared with the blank group, Smad2 mRNA ex－pression in model group significantly increased ( P＜0.05).Vimentin mRNA and Smad2 mRNA expression of low dose group of Taohong Qizhu Ruangan Decoction significantly decreased (P＜0.05).Smad2 mRNA expression of middle dose group of Taohong Qizhu Ruangan Decoction decreased (P＜0.05); TGF－βmRNA, Vimentin mRNA expression of Fuzheng Huayu Capsule group decreased compared with the model group ( P＜0.05).Conclusion:Taohong Qizhu Ruangan Decoction can regulate epithelial－mesenchymal transition through TGF－β/Smad signaling pathway to treat liver fibrosis.