肝乐颗粒对肝纤维化大鼠TGF-β_1/Smad信号通路的调控作用  

Regulation of Ganle Granules on TGF-β_1/Smad Signaling Pathways in Rats with Hepatic Fibrosis

作　　者：张家富[1] 姜辉[1] 高家荣[1] 赵林[2] 刘晓闯[1] 

机构地区：[1]安徽中医药大学第一附属医院,合肥230031 [2]安徽中医药大学药学院,合肥230038

出　　处：《中国实验方剂学杂志》2017年第6期169-174,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：安徽省卫生计生委中医药科研项目(2014zy09)

摘　　要：目的:明确肝乐颗粒对转化生长因子(TGF)-β_1/Smad信号通路的干预作用,探讨其防治肝纤维化的分子学机制。方法:SD大鼠随机分为正常组,模型组,肝乐颗粒(1.5,3.0,6.0 g·kg^(-1))和秋水仙碱(1.0×10^(-4)g·kg^(-1))。采用50%四氯化碳每周2次,连续12周皮下注射的方法,诱导肝纤维化大鼠模型。造模第7周起,给药组分别灌胃给予相应剂量的肝乐颗粒和秋水仙碱,每天1次,连续6周。末次给药8 h后,处置大鼠,取固定部位肝脏组织,苏木素-伊红(HE)和马松(Masson)染色观察肝脏病理组织学损伤程度,免疫组化和蛋白质免疫印迹(Western blot)检测肝组织中Ⅰ型胶原(CollagenⅠ),Smad2,TGF-β_1或TGF-βI型受体(TβRⅠ)蛋白的表达,实时定量荧光PCR(Real-time PCR)检测肝组织中CollagenⅠ,Smad2,TGF-β_1mRNA的表达。结果:与正常组比较,模型组大鼠肝纤维化损伤程度,肝组织中CollagenⅠ,TGF-β_1,Smad2,TβRⅠ蛋白及mRNA的表达明显增加(P<0.01);与模型组比较,肝乐颗粒中、高剂量组不仅可减轻肝组织病理损伤程度,减少胶原沉积,还可显著降低CollagenⅠ,TGF-β_1,Smad2,TβRⅠ蛋白及mRNA的表达(P<0.05,P<0.01)。结论:肝乐颗粒对肝纤维化大鼠具有一定的保护作用,其机制可能与调控TGF-β_1/Smad信号通路,从而抑制HSC活化,减少细胞外基质过度沉积有关。Objective： To study the regulation effect of Ganle granules（ GLG） on transforming growth factor（ TGF-β1） /Smad signaling pathways in rats with hepatic fibrosis,and to explore its possible mechanisms for prevention and treatment of hepatic fibrosis. Method： The SD rats were randomly divided into normal group,model group,GLG（ 1. 5,3. 0,6. 0 g·kg^-1） groups and colchicine（ Col,0. 1 mg·kg^-1） group. The hepatic fibrosis rat models were induced by subcutaneous injection of CCl4（ 50%） twice each week for 12 weeks continuously. Then different doses of GLG（ 1. 5,3. 0,6. 0 g·kg^-1） and colchicines（ 0. 1 g·kg^-1） were administrated via gavage once a day for 6 weeks from week 7 after modeling. 8 h after the last administration,rats were sacrificed,and their liver samples were taken to observe the pathological and histological degree of liver fibrosis by HE and Masson.Moreover,the protein expression levels of Collagen Ⅰ,TGF-β1and Smad2 or TβR Ⅰ in the liver tissues were determined by immunohistochemistry and Western blot. The mRNA expression levels of Collagen Ⅰ,Smad2,and TβRⅠin liver tissues were detected by the real-time fluorescent quantitative PCR. Result： As compared with the normal group,the pathological and histological degree of liver fibrosis,as well as protein and mRNA expression levels of Collagen I,TGF-β1,Smad2,and TβR I in model group were obviously increased in model group（ P〈0. 01）. As compared with model group,GLG middle and high doses not only obviously improved histopathological changes,but also effectively reduced the protein and / or mRNA expression levels of Collagen I,TGF-β1,Smad2,and TβR I（ P〈0. 05,P〈0. 01）. Conclusion： GLG has protective effect on liver fibrosis rats and its mechanism may be related to regulating the TGF-β1/ Smad signaling pathways,inhibiting HSC activation,and reducing excessive extracellular matrix deposition.

关 键 词：肝乐颗粒 肝纤维化 活性生长因子-β1/Smad信号通路 

分 类 号：R285.5[医药卫生—中药学]