PPAR-γ/FAK信号通路在C57BL/6小鼠肺成纤维细胞中的抗纤维化作用  

Antifibrotic effect of PPAR-γ/FAK signaling pathway in C57BL/6 mouse lung fibroblasts

作　　者：龚玲 刘代顺 张龙举 GONG Ling;LIU Dai-shun;ZHANG Long-ju(Department of Respiratory Medicine,the First People’s Hospital of Zunyi(the Third Affiliated Hospital of Zunyi Medical University)/the Respiratory Diseases Institute of Zunyi,Zunyi,Guizhou 563002,China)

机构地区：[1]遵义市第一人民医院呼吸内科(遵义医科大学第三附属医院)/遵义市呼吸疾病研究所,贵州遵义563002

出　　处：《临床肺科杂志》2020年第6期820-824,共5页Journal of Clinical Pulmonary Medicine

基　　金：贵州省中医药管理局(No QZYY-2018-122);贵州省卫生健康委科学技术基金项目(gzwjk2019-1-088)。

摘　　要：目的探讨过氧化物酶体增殖物活化受体-γ(PPAR-γ)配体和粘着斑激酶(FAK)在C57BL/6小鼠肺成纤维细胞中的抗纤维化作用。方法培养C57BL/6小鼠肺成纤维细胞,用TGF-β2 10ng/mL刺激转换为肌纤维母细胞,加入不同浓度PPAR-γ配体罗格列酮(5μmol/L、10μmol/L、20μmol/L及40μmol/L),采用细胞生长计数实验检测罗格列酮对小鼠肺成纤维细胞C57BL/6生长的影响;采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐实验(MTT)检测罗格列酮对小鼠肺成纤维细胞C57BL/6增殖的影响;聚合酶链式反应实验(PCR)检测PPAR-γmRNA、FAK mRNA的转录水平。结果生长计数实验发现罗格列酮抑制肌纤维母细胞生长,在72 h罗格列酮浓度40μmol/L时抑制最明显;MTT实验发现罗格列酮抑制肌纤维母细胞增殖,在72 h罗格列酮浓度40μmol/L时抑制最明显;PPAR-γmRNA转录水平随罗格列酮浓度增加而增加,FAK mRNA的转录水平随罗格列酮浓度增加而减少。结论 PPAR-γ配体罗格列酮可能通过抑制FAK表达进而抑制TGF-β2诱导的肺成纤维细胞纤维化形成。Objective To explore the antifibrotic effect of ligand of peroxisome proliferator activated receptor-gamma(PPAR-γ) and focal adhesion kinase(FAK) on C57 BL/6 mouse lung fibroblasts. Methods C57 BL/6 mouse lung fibroblasts was cultured in vivo, and it was stimulated with TGFβ2(10 ng/mL) into muscle fiber blast cell. Different concentrations of PPAR-γ was combined with different rosiglitazone(5 μmol/L, 10 μmol/L, 20 μmol/L, 40 μmol/L).The effect of rosiglitazone on the growth of mouse lung fibroblasts C57 BL/6 was detected by cell growth count. The effect of rosiglitazone on the proliferation of mouse lung fibroblasts C57 BL/6 was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT). PCR was employed to detect the transcription level of PPAR-γ mRNA and FAK mRNA. Results Growth counting experiment showed that rosiglitazone inhibited the growth of myofibroblasts, and the inhibition was the most obvious at the concentration of 40 μmol/L for 72 h. MTT experiment showed that rosiglitazone inhibited the proliferation of myofibroblasts, and the inhibition was the most obvious at the concentration of 40 μmol/L for 72 h.The transcription level of PPAR mRNA increased with the increase of rosiglitazone concentration. The transcription level of FAK mRNA decreased with the increase of rosiglitazone concentration. Conclusion PPAR-γ ligand rosiglitazone may inhibit the formation of pulmonary fibroblast fibrosis induced by TGF-beta 2 by inhibiting FAK expression.

关 键 词：肺纤维化 过氧化物酶体增殖物激活受体 粘着斑激酶 罗格列酮 转化生长因子-Β2 

分 类 号：R563.9[医药卫生—呼吸系统]