玉屏风多糖对TGF-β1诱导的肝星状细胞增殖的作用及与Smads通路的关系

The effect of proliferation role of Yupingfeng pachy-polysaccharide on TGF-β1-induced rat hepatic stellate cell and the relationship with Smads signaling pathway

目的研究外源性转化生长因子-β1(TGF-β1)对肝星状细胞(HSC)的激活作用,观察玉屏风多糖(YPF-P)对TGF-β1干预的HSC中Smads通路的影响。方法肝星状细胞株HSC-T6用YPF-P(25、50、100 mg/L)处理48 h,同时设空白对照和TGF-β1干预的对照,MTT法检测HSC细胞增殖情况,半定量RT-PCR法检测HSC中Smad2、Smad3、Smad7和结缔组织生长因子(CTGF)mRNA的表达。在此基础上,HSC细胞分别用YPF-P(50 mg/L)、Smads通路阻断剂SIS3(10μmol/ml)、SIS3(10μmol/ml)+YPF-P(50 mg/L)处理48 h,同时设空白对照和TGF-β1干预的对照,Western blot法检测HSC中CTGF蛋白的表达。结果各剂量组YPF-P均能明显抑制TGF-β1干预的HSC增殖,下调Smad2、Smad3、CTGF mRNA的表达,上调Smad7 mRNA的表达。SIS3+YPF-P组CTGF蛋白表达明显下降。结论 TGF-β1明显促进HSC增殖,YPF-P可抑制HSC的活化增殖,其抗纤维化机制可能与阻断HSC中的Smads信号转导通路进而下调CT-GF的表达有关。

Objective To investigate the effect of Yupingfeng pachy-polysaccharide（YPF-P） on Smads signaling pathway in transforming growth factor-β1（TGF-β1） induced rat hepatic stellate cells（HSC） and to explore the activation of TGF-β1 on HSC.Methods HSC were dealed with YPF-P（25,50 and 100 mg/L） for 48 h,and treated with nothing or TGF-β1.MTT assay was used to evaluate the proliferation of HSC.The mRNA expressions of Smad2,Smad3,Smad7 and connective tissue growth factor（CTGF） in HSC were detected by RT-PCR.Based on the above,HSC were dealed with YPF-P（50 mg/L）,blocking agent SIS3（10 μmol/ml）,YPF-P（50 mg/L） and SIS3（10 μmol/ml） for 48 h,and meanwhile compared with control and TGF-β1-induced group.The expression of CTGF protein was observed by Western blot.Results The different doses of YPF-P inhibited the proliferation of HSC and down-regulated the expressions of Smad2,Smad3,and CTGF mRNA,but up-regulated Smad7 mRNA in HSC.Conclusion TGF-β1 could promote the proliferation of HSC,but YPF-P could inhibit the activation and proliferation of HSC.The mechanism might be associated with inhibiting the Smads signaling pathway and down-regulating the expression of CTGF protein in HSC.