TGF-β1诱导的肺泡上皮细胞间质转化及其机制探讨  

Exploration of the process and mechanism for the epithelial mesenchymal cell transformation of pulmonary alveoli induced by TGF-β1

作　　者：黄振杰[1] 曾彤华[1] 蔡文华[1] 欧盛敬[1] 陈文海[1] 李斐[1] 

机构地区：[1]广西北海市人民医院呼吸内科,536000

出　　处：《中华肺部疾病杂志（电子版）》2016年第3期292-296,共5页Chinese Journal of Lung Diseases(Electronic Edition)

基　　金：广西卫生厅科研自筹基金资助项目(Z2011063);北海市科研自筹基金项目资助(北科合201109002)

摘　　要：目的探讨转化生长因子β1(TGF-β1)诱导的肺泡上皮细胞-间质细胞转化(EMT)过程及信号传导通路。方法体外培养肺腺癌细胞系A549细胞,以TGF-β1进行干预;采用Western blot免疫印迹法检测干预前后细胞标志物蛋白及信号传导蛋白P-Smad2/3、Snail1、Snail2的表达;采用RT-PCR检测干预前后细胞标志物mRNA表达;采用倒置相差显微镜观察细胞形态学变化。结果 TGF-β1干预后,随着时间的延长,A549细胞上皮细胞标志物E-cad和CK19蛋白及mRNA表达下调(P<0.05);间质细胞标志物α-SMA、Vimentin蛋白及mRNA表达上调(P<0.05);信号传导蛋白P-Smad2/3、Snail1表达上调(P<0.05);倒置相差显微镜观察TGF-β1干预后A549细胞由干预前的鹅卵石状变为梭形,如同肌纤维母细胞。结论 TGF-β1可诱导肺泡上皮细胞向间质细胞转化,其机制可能与P-Smad2/3、Snail1信号传导通路有关;TGF-β1可能是通过诱导肺泡上皮细胞间质转化的细胞效应,最终导致肺泡上皮细胞凋亡和纤维组织形成。Objective To explore the process of Epithelial mesenchymal cell transformation（ EMT） of pulmonary alveoli induced by transforming growth factorβ1（ TGF-β1） and its signal transduction pathways.Method Cultivated the adenocarcinoma cells of lung A549 in vitro, intervented by TGF-β1, using Westernblot method to detect the expression of the cell markers protein and signal transduction protein PSmad2 / 3,Snail1,Snail2 before and after the intervention,using RT-PCR to detect the mRNA expression of the cell markers before and after the intervention. Observed the cell morphology by inverted phase contrast microscope. Results After the TGF-β1 intervention,with the extension of time,the protein and mRNA expression of the A549 cells epithelial markers E-cad and CK19 were low-regulation（ P〈0. 05）; The protein and mRNA expression level of the interstitial cell markers α-SMA,Vimentin were up-regulation（ P〈0. 05）;The expression of the signal transduction protein P-Smad2 / 3,Snail1 were up-regulation（ P〈0. 05）; Inverted phase contrast microscope observation： The A549 cells turn into spindle shape of after TGF-β1 intervention from pebble shape pre-intervention,like a muscle fiber mother cell. Conclusion TGF-β1 can induce alveolar epithelial turn into mesenchymal cells and its mechanism may be related to P-Smad2 / 3, Snail1 signal transduction pathways. TGF-β1 may result in the apoptosis of alveolar epithelial cells and fibrogenesis just by inducing epithelial-mesenchymal transition-like cellular response.

关 键 词：肺纤维化 转化生长因子Β1 A549细胞 上皮细胞间质转化 信号传导通路 

分 类 号：R563[医药卫生—呼吸系统]