肝复健方对肝纤维化大鼠TGF-β/Smad信号通路的影响

目的 观察以"浊毒"立论的肝复健方对猪血清诱导的大鼠肝纤维化(HF)肝脏转化生长因子-β1(TGF-β1)及其Ⅱ型受体(TβRⅡ) mRNA水平和Smad2/3、Smad4、Smad7蛋白表达的影响.方法 SD大鼠腹腔注射猪血清0.5 mL,每周2次,连续8周制备HF模型,每天灌胃分别给予不同剂量的肝复健方(15.0、22.5、30、0 g/kg)和秋水仙碱(0.154 g/kg)治疗至12周末.实验结束后,RT-PCR法检测肝组织TGF-β1及其受体TβRⅡmRNA的表达,Weston-blot法检测肝组织Smad2/3、Smad4、Smad7蛋白的表达.结果 与正常对照组相比,模型组TGF-β1、TβRⅡ及Smad2/3、Smad4表达明显增强,Smad7表达明显下降;经肝复健方治疗后大鼠肝脏TGF-β1及其受体TβRⅡ表达均比模型组减弱,Smad2/3、Smad4的表达减弱而Smad7蛋白的表达增强.结论 肝复健方能抑制TGF-β1及其受体TβRⅡ mRNA的表达,下调Smad2/3、Smad4蛋白表达,上调Smad7蛋白的表达.肝复健方对大鼠HF肝脏TGF-β/Smad信号通路有明显的影响,这可能是其治疗HF的机制之一.

Influences of Ganfujian Formula on expressions of TGF-β and Smad signal pathway in rats with hepatic fibrosis

Objective To observe the influences of Ganfujian Formula on the expressions of transforming growth factor-β1 (TGF-β1), TβRII-mRNA, Smad2/3, Smad4 and Smad7 in rats with hepatic fibrosis induced by porcine serum.Methods The hepatic fibrosis model was established in SD rats by the intraperitoneal injection of porcine serum (0.5 mL, twice a week for 8 weeks). The rats then were given Ganfujian Formula orally in different doses (1.50 g/kg, 2.25 g/kg and 3.0 g/kg) and colchicines (0.154 g/kg) respectively until the end of 12th week. The expressions of TGF-β1 and TβRII-mRNA were detected by using RT-PCR and the expressions of Smad2/3, Smad4 and Smad7 were detected by applying Western blotting method.Results In the model group, compared with the normal control group, the expressions of TGF-β1 and TβRII-mRNA, Smad2/3 and Smad4 increased significantly and Smad7 expression decreased significantly. After the treatment with Ganfujian Formula the expressions of TGF-β1 and TβRII-mRNA were reduced in the treatment group compared with the model group. Ganfujian Formula regulated downward the expressions of Smad2/3 and Smad4, and regulated upward Smad7 expression at the same time.Conclusion Ganfujian Formula can inhibit the expressions of TGF-β1 and TβRII-mRNA, regulates downward the expressions of Smad2/3 and Smad4, and regulates upward Smad7 expression. It may be its one of the mechanisms for curing hepatic fibrosis that it has an obvious influence on hepatic TGF-β and Smad signal pathway.