论文部分内容阅读
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 clarify the effect of Ganle Granule on TGF-β1 / Smad signaling pathway and to explore its molecular mechanism of prevention and treatment of hepatic fibrosis. Methods: SD rats were randomly divided into four groups: normal group, model group, Ganle Granule (1.5,3.0,6.0 g · kg -1) and colchicine (1.0 × 10 -4 g · kg ~ (-1) -1)). The model of hepatic fibrosis was induced by subcutaneous injection of 50% carbon tetrachloride twice a week for 12 weeks. From the 7th week after model establishment, the rats in the treatment group were given gavage respectively with gavage and colchicine, once a day for 6 weeks. Eight hours after the last administration, the rats were sacrificed and the histopathological damage, immunohistochemistry and Western blot were observed by staining the liver, HE and Masson, The expression of collagen Ⅰ, Smad2, TGF-β 1 or TGF-β 1 receptor (TβRⅠ) in liver tissue were detected by real-time PCR. The expressions of collagenⅠ, Smad2 and TGF- β 1 mRNA expression. Results: Compared with the normal group, the expression of CollagenⅠ, TGF-β_1, Smad2, TβRⅠprotein and mRNA in the model group was significantly increased (P <0.01), compared with the model group, The medium and high dose groups not only alleviated the degree of pathological damage of liver tissue and decreased collagen deposition, but also significantly decreased the expressions of CollagenⅠ, TGF-β_1, Smad2 and TβRI (P <0.05, P <0.01). CONCLUSION: Ganle Granule has a protective effect on rats with liver fibrosis, and its mechanism may be related to the regulation of TGF-β1 / Smad signaling pathway, thereby inhibiting HSC activation and reducing extracellular matrix over-deposition.