鳖甲煎改良方对大鼠肝纤维化的作用及其机制研究

Effect of modified turtle shell decoction on hepatic fibrosis in rats and its mechanism

目的探讨鳖甲煎改良方对大鼠肝纤维化(hepatic fibrosis,HF)的作用及其机制。方法 SD雄性大鼠90只,随机分6组,每组15只,其中以40%CCl4处理(3 ml/kg)8周复制中毒肝纤维化模型(模型组),复制模型的同时分别用鳖甲煎改良方高剂量组28.4 g/(kg.d)、中剂量组14.2 g/(kg.d)、低剂量组7.1 g/(kg.d)灌胃治疗并且以复方鳖甲软肝片灌胃0.6 g/(kg.d)作阳性对照组治疗6周。以复制模型同样方式与用量于右后腿皮下注射橄榄油及生理盐水灌胃(10 ml/kg.d)作空白对照组。苏木精-伊红(HE)染色观察各组8周后肝纤维化变化程度,RT-PCR检测其转化生长因子β1(transforming growth factor beta one,TGF-β1)、Smad3及Smad7变化,免疫组化(S-P法)观测TGF-β1、smad3、smad7表达情况。结果 8周后,与模型组比较,鳖甲煎改良方高、中、低剂量组和复方鳖甲软肝片组肝小叶结构破坏明显减轻,部分肝细胞脂肪变性,炎性细胞浸润与少量胶原纤维形成;肝组织TGF-β1与Smad3的mRNA及蛋白表达显著减少(P<0.01),smad7 mRNA及蛋白表达明显增加(P<0.01)且鳖甲煎改良方高、中、低剂量组和复方鳖甲软肝片组疗效并无显著差异。结论鳖甲煎改良方能显著改善大鼠肝纤维化,其作用机制可能与鳖甲煎改良方调控TGF-β/smad信号通路有关。

Objective To study the effect of modified turtle shell decoction（MPTSD） on hepatic fibrosis in rats and its mechanism.Methods Ninety male SD rats were randomly divided into 6 groups,15 rats in each group.A toxic hepatic fibrosis of rats was reproduced with 40% carbon tetrachloride（CCl4）（3 ml/kg） for 8 weeks.Fifteen rats were treated for 6 weeks with MPTSD at 28.4 g/（kg·d） as a high dose group,at 14.2 g/（kg·d） as a medium dose group,and at 7.1 g/（kg·d） as a low-dose group.Rats treated with compound MPTSD soft tablets at 0.6 g/（kg·d） served as a positive control group and those treated with olive oil at 10 ml/（kg·d） served as a blank control group.Changes of HF in different groups were observed with hematoxylin-eosin（HE）staining 8 weeks after treatment.Expression of transforming growth factor beta 1（TGF-β1）,Smad3 and Smad7 was detected by reverse transcriptase polymerase chain reaction（RT-PCR） and immunohistochemistry staining.Results Eight weeks after treatment,the destruction of hepatic lobules in high,medium and low dose MPTSD groups and positive control group were remarkably alleviated with,fatty degeneration of some liver cells,infiltration of inflammatory cells and formation of few collagen fibers.The mRNA and protein expression level of TGF-β1 and Smad3 was significantly lower while the expression level of Smad7 was remarkably higher in high,medium and low dose MPTSD groups and positive control group than in blank control group（P0.01）.No significant difference was observed in high,medium and low dose MPTSD groups and positive control group.Conclusion MPTSD can significantly improve CCl4-induced hepatic fibrosis in rats by regulating the TGF-β/smad signal pathway.