脐带间质干细胞来源的外泌体抑制TGF-β_1诱导的NRK-52E细胞上皮-间质转化  

Exosome derived from human umbilical mesenchymal stem cells inhibited TGF-β_1-induced epithelial-mesenchymal transition of renal proximal tubular epithelial cells

作　　者：俞静[1] 张荣雪[1] 贾浩源[1] 王娟娟[1] 纪成[1] 许文荣[1] 钱晖[1] 

机构地区：[1]江苏大学医学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2017年第4期277-281,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81272481);江苏省高校自然科学研究重大项目(15KJA320001);江苏省"333工程"科研项目(BRA2015399);江苏高校优秀科技创新团队项目[苏教科(2013)10号]

摘　　要：目的:探讨脐带间质干细胞来源的外泌体(exosome from human umbilical mesenchymal stem cells,huc MSCex)对转化生长因子β_1(TGF-β_1)诱导的NRK-52E细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)的影响及其机制。方法:体外培养大鼠肾小管上皮NRK-52E细胞,TGF-β_1诱导使其发生EMT作为诱导组,未经任何处理的NRK-52E细胞为对照组,TGF-β_1处理24 h后加入huc MSC-ex共培养48 h为干预组。倒置显微镜下观察各组细胞形态;蛋白质印迹法检测细胞中E-钙黏蛋白、N-钙黏蛋白、α-平滑肌肌动蛋白(α-SMA)、I型胶原α1链(Col1A1)和TGF-β_1等蛋白表达,qRT-PCR检测miR-199a表达。结果:TGF-β_1诱导24 h后,细胞由卵圆形变为长梭形,且细胞密度降低。给予huc MSC-ex干预后,细胞形态恢复。与对照组相比,诱导组E-钙黏蛋白表达降低,N-钙黏蛋白,α-SMA、Col1A1和TGF-β_1表达升高;干预组相关蛋白表达较诱导组发生逆转;诱导组中miR-199a表达下调,而huc MSC-ex干预组表达增加。结论:huc MSC-ex通过上调NRK-52E细胞miR199a表达逆转由TGF-β_1诱导的EMT作用。Objective： To investigate the effect of exosome derived from human umbilical mesenchymal stem cells（ huc MSC-ex） on the epithelial-mesenchymal transition（ EMT） of NRK-52 E cells induced by TGF-β1. Methods： Rat renal proximal tubular epithelial NRK-52 E cells were cultured in vitro. NRK-52 E cells without any treatment were selected as control group. NRK-52 E cells were induced by TGF-β1for 24 h and were named induction group. The induced cells were co-cultured with exosomes for 24 h and selected as intervention group. The cellular morphology in three groups were observed by microscope.Western blotting were used to determine the expression of EMT indicator,such as E-cadherin,N-cadherin,α-SMA,TGF-β1and Col1A1. qRT-PCR was used to detect the expression level of miR-199 a in groups. Results： With the stimulation of TGF-β1,the morphology of NRK-52 E cells changed and the process of EMT happened in NRK-52 E cells. However,after co-cultured with exosome,the morphology of cells returned to normal and the EMT phenotype had ameliorated. Compared with the control group,the expression of E-cadherin were decreased,while the expression of N-cadherin,α-SMA,Col1A1,TGF-β1were increased in induction group; compared with the induction group,the expression of all the proteins were restored in the intervention group. Compared with the control group,miR-199 a level decreased in the induction group and restored in the intervention group. Conclusion： Huc MSC-ex could suppress the process of EMT induced by TGF-β1in NRK-52 E cells via upregulating miR-199 a expression.

关 键 词：脐带间质干细胞 外泌体 上皮-间质转化 MIRNA 

分 类 号：R393[医药卫生—基础医学]