加味茵陈四逆汤含药血清对TGF-β_1干预的大鼠肝星状细胞TGF-β_1/Smads通道及胶原蛋白的影响

Effect of Drug Serum of Modified Yinchen Sini Tang on TGF-β_1/ Smads Signaling Pathways and Collagen of TGF-β_1-induced Rat Hepatic Stellate Cells

目的:探讨加味茵陈四逆汤含药血清对转化生长因子-β1(TGF-β1)干预的大鼠肝星状细胞TGF-β1/Smad通道相关蛋白及胶原蛋白3(Collagen3)生成量的影响。方法:以大鼠肝星状细胞HSC-T6为研究对象,将细胞分为空白组,模型组,阳性药物组(甲强龙,10%含药血清),加味茵陈四逆汤高、中、低剂量组(10%含药血清),共6组。除空白组外,其余均给予TGF-β1(10 g·L-1)并采用相应10%的含药血清培养,并采用蛋白质免疫印迹(Western blot)的方法检测各研究组含药血清对大鼠肝星状细胞HSC-T6 Smad3,Smad7,Collagen3蛋白表达的影响。结果:与空白组比较,模型组HSC-T6细胞Smad3,Smad7,Collagen3蛋白表达升高,Collagen3蛋白表达升高较为明显(P<0.05);与模型组比较,加味茵陈四逆汤低剂量明显降低Smad3蛋白表达(P<0.05),加味茵陈四逆汤中剂量明显降低Collagen3蛋白表达(P<0.05)。结论:加味茵陈四逆汤含药血清使Smad3,Collagen3蛋白表达下调,Smad7未见明显变化,提示本药在离体条件下可以延缓肝纤维化的进程。

Objective： To investigate the effect of drug serum of modified Yinchen Sini Tang on transforming growth factor（ TGF）-β1/ Smads signaling pathways and Collagen3 of TGF-β1-induced rat hepatic stellate cells（ HSC）. Method： With rat hepatic stellate cells as the study objects,the cells were divided into blank group,model group,positive drug group（ methylprednisolone,10% drug serum）,modified Yinchen Sini Tang high dose,middle dose and low dose groups（ 10% drug serum）. All the other rats except those in blank group were given with TGF-β1（ 10 g·L^- 1） and corresponding 10% drug serum culture was used,the effects of the drug serum on protein expression levels of Smad3,Smad7,Collagen3 in HSC-T6 cells were detected by using Western blot assay. Result： As compared with the blank group,protein expression levels of Smad3,Smad7,Collagen3 in HSC-T6 cells were increased in model group,and the increase of Collagen3 expression was more obvious（ P 〈0. 05）. As compared with the model group,the protein expression level of Smad3 was significantly decreased in modified Yinchen Sini Tang low dose group（ P 〈0. 05）,the protein expression level of Collagen3 was significantly decreased in modified Yinchen Sini Tang middle dose group（ P 〈0. 05）. Conclusion： Drug serum of modified Yinchen Sini Tang could down-regulate the protein expression levels of Smad3 and Collagen3,while there was no significant change in Smad7,suggesting that the drug can delay the process of liver fibrosis under the in vitro conditions.