【摘要】 目的:明确凋亡抑制蛋白c-FLIP(L)在肺纤维化过程中的作用,探究其异常表达参与肺纤维化发病的分子机制。方法:构建博来霉素诱导的小鼠肺纤维化模型,对正常鼠与模型鼠的肺组织切片进行HE和Masson染色观察病理变化,并采用免疫组化分析c-FLIP(L)及上皮-间质转化(epithelial-mesenchymal transition,EMT)标志分子E-cadherin的表达。构建表达c-FLIP(L)的稳定细胞株,qRT-PCR检测EMT标志分子E-cadherin、N-cadherin及Vimentin的mRNA表达。采用转化生长因子(transforming growth factor,TGF)-β1诱导细胞发生EMT,通过Smad报告基因检测和Western blot分析c-FLIP对TGF-β1诱导EMT发生的影响。结果:c-FLIP(L)表达水平在肺纤维化组织中明显升高,与E-cadherin表达呈负相关性。C-FLIP(L)能促进肺上皮细胞的EMT表型,并促进TGF-β1诱导的Smad信号通路激活,而敲减c-FLIP(L)表达能阻滞TGF-β1诱导的EMT进程。结论:c-FLIP(L)在肺纤维化过程中高表达能促进EMT发生,是肺纤维化病程发展的促进因素之一。更多还原

【Abstract】 Objective:This study aims to explore the effect of c-FLIP(L)in pulmonary fibrosis and its pathological mechanism.Methods:Based on bleomycin(BLM)induced idiopathic pulmonary fibrosis in mice,the expressions of c-FLIP and E-cadherin were analyzed by IHC staining. Further investigations in A549 cells overexpressing c-FLIP(L),the expression levels of E-cadherin,Ncadherin and Vimentin mRNA were examined by qRT-PCR,and Smad activation induced by transforming growth factor(TGF)-β1 was detected by luciferase reporter assay and Western blot. Results:The increased c-FLIP expression was observed and associated with a decrease of E-cadherin expression. C-FLIP(L)overexpression resulted in the changes on E-cadherin,N-cadherin and Vimentin expressions in A549 cells. Furthemore,overexpression of c-FLIP(L)enhanced TGF-β-induced Smad activation,and knocking down c-FLIP(L)blocked the TGF-β1-induced EMT progress. Conclusion:C-FLIP(L)may be a promoting factor in the development of fibrosis via regulating EMT progress.更多还原