学术论文

      脐带间质干细胞来源的外泌体抑制TGF-β1诱导的NRK-52E细胞上皮-间质转化

      Exosome derived from human umbilical mesenchymal stem cellsinhibited TGF-β1-induced epithelial-mesenchymal transition ofrenal proximal tubular epithelial cells

      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, hucMSC-ex)对转化生长因子β1(TGF-β1)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-β1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-β1处理24 h后加入hucMSC-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-β1等蛋白表达,qRT-PCR检测miR-199a表达.结果: TGF-β1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hucMSC-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-β1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hucMSC-ex干预组表达增加.结论: hucMSC-ex通过上调NRK-52E细胞miR199a表达逆转由TGF-β1诱导的EMT作用.
      Abstract:
      Objective: To investigate the effect of exosome derived from human umbilical mesenchymal stem cells(hucMSC-ex) on the epithelial-mesenchymal transition (EMT) of NRK-52E cells induced by TGF-β1.Methods: Rat renal proximal tubular epithelial NRK-52E cells were cultured in vitro.NRK-52E cells without any treatment were selected as control group.NRK-52E cells were induced by TGF-β1 for 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-β1 and Col1A1.qRT-PCR was used to detect the expression level of miR-199a in groups.Results: With the stimulation of TGF-β1, the morphology of NRK-52E cells changed and the process of EMT happened in NRK-52E 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-β1 were 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-199a level decreased in the induction group and restored in the intervention group.Conclusion: HucMSC-ex could suppress the process of EMT induced by TGF-β1 in NRK-52E cells via upregulating miR-199a expression.
      Author: YU Jing ZHANG Rong-xue JIA Hao-yuan WANG Juan-juan JI Cheng XU Wen-rong QIAN Hui
      作者单位: 江苏大学医学院,江苏 镇江,212013
      年,卷(期): 2017, 27(4)
      分类号: R393
      在线出版日期: 2017年7月31日
      基金项目: 国家自然科学基金资助项目,江苏省高校自然科学研究重大项目,江苏省"333工程"科研项目,江苏高校优秀科技创新团队项目