肝X受体激动剂T0901317抑制TGF-β1诱导的人正常肺成纤维细胞α-SMA的表达  

Liver X receptor agonist T0901317 inhibits TGF-β1-induced α-SMA expression in normal human lung fibroblasts

作　　者：任敦强[1] 刘明[1] 郭永忠[1] 马冉[1] 钟南山[1,2] 

机构地区：[1]广州医学院第一附属医院广州呼吸疾病研究所呼吸疾病国家重点实验室,广东广州510120 [2]中国工程院

出　　处：《南方医科大学学报》2011年第5期744-748,共5页Journal of Southern Medical University

基　　金：国家"973"项目(2009CB522100);广东省自然科学基金重点项目(07117506)~~

摘　　要：目的探讨肝X受体激动剂T0901317对TGF-β1诱导的人正常肺成纤维细胞α-平滑肌肌动蛋白(α-SMA)表达的影响。方法组织块贴壁法培养人正常肺成纤维细胞,波形蛋白和角蛋白免疫细胞化学鉴定。T0901317和/或TGF-β1刺激人正常肺成纤维细胞后,分别应用RT-PCR,Western blot和免疫荧光观察α-SMA mRNA和蛋白水平的表达。结果人肺成纤维细胞表达波形蛋白而不表达角蛋白。RT-PCR,Western blot,免疫荧光结果均显示正常肺成纤维细胞基态下组成性表达少量的α-SMA,TGF-β15ng/ml即可以诱导α-SMA mRNA和蛋白表达的明显上调,而T09013175μg/ml时可以明显抑制这种表达的上调。结论肝X受体激动剂T0901317可以在基因和蛋白水平抑制TGF-β1诱导的体外人正常肺成纤维细胞α-SMA表达的上调,可能具有潜在抗纤维化应用价值。Objective To investigate the effect of liver X receptor agonist T0901317 on transforming growth factor-β1（TGF-β1）-induced expression of α-smooth muscle actin（α-SMA） in normal human lung fibroblasts.Methods Primary normal human lung fibroblast isolated from the lung specimens of lung cancer patients by explant culture technique were identified with immunostaining for vimentin and keratin.The cells in passages 4 to 10 were treated with T0901317 and/or TGF-β1,and RT-PCR,Western blotting and immunoflueresence assay were used to detect α-SMA expression in the fibroblasts.Results Lung fibroblast expressed vimentin but not keratin.The results of RT-PCR,Western blotting and immunoflueresence assay all showed that normal human lung fibroblasts constitutively expressed α-SMA under baseline condition,and TGF-β1 at 5 ng/ml induced a significant upregulation of α-SMA both at the mRNA and protein levels.Liver X receptor agonist T0901317（5 μg/ml） significantly inhibited TGF-β1-induced upregulation of α-SMA expression.Conclusion Liver X receptor agonist T0901317 can inhibit the upregulation of α-SMA in normal human lung fibroblasts induced by TGF-β1,suggesting the potential value of liver X receptor agonist in the treatment of lung fibrosis.

关 键 词：转化生长因子-β1 人肺成纤维细胞 肝X受体 T0901317 α-平滑肌肌动蛋白 

分 类 号：R563.9[医药卫生—呼吸系统]