加味茵陈四逆汤对肝纤维化小鼠Smad7及Ⅰ、Ⅲ型胶原蛋白表达的影响

Effect of Jiawei Yinchen Sini Decoction on the Production of Smad7 and CollagenⅠ,Ⅲ in Carbon Tetrachloride Induced Hepatic Fibrosis Model of Mice

目的:探讨预防性使用加味茵陈四逆汤对肝纤维化小鼠Smad7和Ⅰ型胶原蛋白(CollagenⅠ)、Ⅲ型胶原蛋白(CollagenⅢ)基因表达的影响。方法:雄性ICR小鼠48只,随机分为正常对照组、模型组、秋水仙碱组及加味茵陈四逆汤高、中、低剂量组,每组8只。采用腹腔注射30%CCl4(1.5 mg/kg,溶解于橄榄油)建立肝纤维化模型,同时给予药物灌胃处理。用药14 d后,计算肝脏指数;试剂盒检测血清透明质酸(HA)、层粘连蛋白(LN)水平;各组行HE染色观察肝组织病理改变并进行肝纤维化分级;RT-q PCR法检测肝组织Smad7、CollagenⅠ、CollagenⅢmRNA表达。结果:(1)加味茵陈四逆汤中、低剂量组肝脏指数和HA、LN水平较模型组显著降低(P<0.05);(2)加味茵陈四逆汤各剂量组肝纤维化较模型组减轻(P<0.05);(3)与模型组比较,各给药组CollagenⅠmRNA表达显著降低(P<0.05),除加味茵陈四逆汤高剂量组外,各给药组Smad7 mRNA表达显著升高(P<0.05),除加味茵陈四逆汤低剂量组外,各给药组CollagenⅢmRNA表达显著降低(P<0.05)。结论:加味茵陈四逆汤可上调肝纤维化小鼠Smad7表达,抑制CollagenⅠ、CollagenⅢ表达,发挥对肝纤维化的抑制作用。

Objective： To explore the preventive effects of Jiawei Yinchen Sini Decoction on the gene expression of Smad7 and CollagenⅠ,Ⅲ in mice with hepatic fibrosis. Methods： 48 male ICR mice were randomly divided into normal group,model group,colchicine group,JWYSN high dose group,medium dose group and low dose group,with 8 mice in each group. The hepatic fibrosis model was established by intraperitoneal injection of 30% CCl4（ 1. 5 mg/kg,dissolved in olive oil）. At the same time,the mice were treated with intragastric adminstration of drugs. After 14 days,the liver index was calculated; the levels of serum hyaluronic acid（ HA） and laminin protein（ LN） were detected by Enyme-linked immunosorbent assay; the pathological fibrosis change was observed by performing HE staining and was made a grading of hepatic fibrosis. mRNA expression of Smad7 and Collagen Ⅰ,Ⅲ were detected by RT-q PCR analysis. Results：（ 1） Compared with model group,the liver index and levels of HA and LN in JWYSN medium dose group and low dose group were obviously decreased（ P〈0. 05）.（ 2） The pathological results showed that,compared with model group,the hepatic fibrosis were significantly relief in different dose of JWYSN group（ P〈0. 05）.（ 3） The expressions of Collagen Ⅰ mRNA in each treatment group were significantly lower than model group（ P〈0. 05）; In addition to JWYSN high dose group,the expressions of Smad7 mRNA in each treatment group increased significantly（ P〈0. 05）; In addition to JWYSN low dose group,the expression of CollagenⅢmRNA in each treatment group decreased significantly（ P〈0. 05）. Conclusion： Jiawei Yinchen Sini decoction can up-regulate the expressions of Smad7 mRNA and inhibit expressions of mRNA and CollagenⅢmRNA in mice with hepatic fibrosis,thus attenuate the development of hepatic fibrosis.