靶向沉默HIF-1α信号通路抑制百草枯中毒诱导的肺泡上皮细胞上皮间质转化  

Paraquat Promotes Epithelial-mesenchymal Transition of Alveolar Epithelial Cells through HIF-1α Signaling Pathway

作　　者：王晓芬[1] 李福祥[1] 黎俊雅 朱忠立[1] 祝国芸 WANG Xiao-fen;LI Fu-xiang;LI Jun-ya;ZHU Zhong-li;ZHU Guo-yun(Departent of Critical Care,Chengdu General Hospital,Chengdu,Sichuan,610083,China)

机构地区：[1]成都军区总医院重症医学科,四川成都610083

出　　处：《现代生物医学进展》2018年第13期2419-2423,2430,共6页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81170281)

摘　　要：目的:探讨HIF-1α信号通路在百草枯(paraquat,PQ)诱导大鼠Ⅱ型肺泡上皮细胞上皮间质转化(Epithelial-mesenchymal transition,EMT)中的作用机制。方法:使用20μmol/L浓度的百草枯溶剂对大鼠Ⅱ型肺泡上皮RLE-6TN细胞干预24 h,随后在倒置光学显微镜观察各组细胞形态学变化;用real-time PCR与Western blot法检测RLE-6TN细胞中HIF-1α、上皮表型标记蛋白E-cadherin及间质表型标记蛋白Vimentin的表达,Transwell侵袭实验检测各处理组细胞侵袭能力的改变;使用HIF-1α靶向si RNA抑制其表达后,进一步采用RT-PCR和Western blot检测HIF-1α、E-cadherin和Vimentin的表达水平,Transwell法检测细胞侵袭能力变化。结果:体外百草枯溶液可显著诱导大鼠Ⅱ型肺泡上皮细胞RLE-6TN细胞HIF-1α表达升高和上皮间质转化的发生,同时细胞的体外侵袭能力也增强。靶向沉默HIF-1α基因后,百草枯诱导的上皮间质转化过程被逆转,同时细胞侵袭能力显著减弱。结论:百草枯通过调控HIF-1α信号通路来诱导RLE-6TN细胞上皮间质转化的发生,进而促进肺纤维化的形成。Objective： To investigate the effect of HIF-1α signaling pathway on paraquat（PQ）-induced epithelial-mesenchymal transition（EMT） in rat alveolar type II cells（RLE-6 TN） and explore the underlying molecular mechanisms. Methods： RLE-6 TN cells were treated by 20 μmol/L PQ, then the morphology was observed by invert light microscope; RT-PCR and Western blot were performed to detect the expression level of EMT related markers, E-cadherin and vimentin as well as HIF-1α signaling. Then the Transwell invasion assays was performed to detect the ability of cell invasion. Results： PQ was able to induce the transition of RLE-6 TN cells from epithelial morphology to fibroblast-like morphology, associated with the acquisition of migratory properties. Phenotypically, PQ induced-EMT was characterized by loss of epithelial cell markers including E-cadherin, while upregulation of mesenchymal cell markers including vimentin, concurrent with the activation of HIF-1α signaling pathway. Furthermore, knockdown of HIF-1α by using specific si RNA could reverse PQ triggered EMT process and attenuated cell migration ability. Conclusion： PQ promoted EMT in rat alveolar type II cells（RLE-6 TN） by upregulating the expression of HIF-1α.

关 键 词：百草枯 肺纤维化 缺氧诱导因子-1A 上皮-间质转化 

分 类 号：R-33[医药卫生] R595.4