【摘要】 [目的]探讨糜酶抑制剂(Chy-I)在体外对肝星状细胞(HSC)增殖及肝纤维化相关基因表达的影响及作用机制.[方法]取HSC-T6细胞,分为对照组(DMEM)、模型组(TGF-β1)和Chy-I大、中、小剂量(TGF-β1+100 g/L Chy-I,TGF-β1+10 g/L Chy-I,TGF-β1+1 g/L Chy-I)组,检测Chy-I对HSC-T6细胞的增殖抑制率,采用Western blot法检测纤维化相关因子α-SMA及TGF-β1蛋白表达水平,应用实时定量PCR法检测α-SMA及I型胶原mRNA的表达情况.[结果]与模型组比较,Chy-I各剂量组HSC-T6细胞增殖明显受到抑制(P<0.05,P<0.01),且呈剂量依赖性;Western blot法检测结果显示,与模型组比较,Chy-I各剂量组HSC-T6细胞中α-SMA及TGF-β1蛋白表达明显受到抑制,亦呈剂量依赖性,Chy-I各组HSC-T6细胞内α-SMA及I型胶原mRNA表达水平明显降低.[结论] Chy-I在体外对HSC发挥抗肝纤维化作用,其机制认为可能与抑制HSC增殖水平及肝纤维化相关因子的表达有关系.更多还原

【Abstract】 OBJECTIVE To investigate the effects of chymase inhibitor(Chy-I) on the proliferation of hepatic stellate cells(HSC) and the expression of hepatic fibrosis-related genes in vitro and its mechanism. METHODS HSC-T6 cells were cultured and divided into control group(DMEM), model group(TGF-β1) and Chy-I high, medium and low dose groups(TGF-β1+100 g/L Chy-I,TGF-β1+10 g/L Chy-I,TGF-β1+1 g/L Chy-I). The proliferation inhibition rate of Chy-I on HSC-T6 cells was detected. Western blot was used to detect the protein expression levels of fibrosis-related factors α-SMA and TGF-β1. Real-time PCR was used to detect the mRNA expressions of α-SMA and collagen I. RESULTS Compared with model group, the proliferations of HSC-T6 cells in all Chy-I groups were significantly inhibited(P<0.05,P<0.01), and in a dose-dependent manner. The results of Western blot showed that the protein levels of α-SMA and TGF-β1 of HSC-T6 cells in each Chy-I group were significantly inhibited as compared with the model group, and it was also in a dose-dependent manner. The mRNA expressions of α-SMA and collagen I of HSC-T6 cells in each Chy-I group were significantly decreased. CONCLUSION Chy-I played the role of anti-hepatic fibrosis on HSC in vitro, and its mechanism might be related to the inhibition of HSC proliferation and the expression of hepatic fibrosis-related factors.更多还原