大鼠骨髓间质干细胞来源的外切体抑制顺铂诱导的肾小管上皮细胞凋亡  

Exosome derived from rat bone marrow mesenchymal stem cells inhibits the apoptosis in cisplatin-induced renal proximal tubular epithelial cells

作　　者：祝源[1] 杨芳[1] 张娇[1] 薛建国[1] 尹磊[1] 俞静[1] 贾浩源[1] 纪成[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）》2015年第3期203-206,211,共5页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(81272481);江苏高校优秀科技创新团队资助项目[苏教科(2013)10号];江苏省医学领军人才与创新团队项目(LJ201117);江苏省高等学校大学生创新创业训练计划项目(201310299058Y)

摘　　要：目的:观察大鼠骨髓间质干细胞(mesenchymal stem cells,MSC)来源的外切体(exosome)对顺铂诱导的肾小管上皮细胞凋亡的影响。方法:原代培养大鼠骨髓MSC并提取外切体鉴定。构建顺铂损伤NRK-52E细胞模型,损伤组为5μmol/L顺铂处理6 h后正常营养液培养48 h,处理组为顺铂损伤后以100μg/m L外切体共培养48 h。未经任何处理的NRK-52E细胞为对照组。TUNEL-FITC,凋亡双染流式细胞术,蛋白质免疫印迹技术检测各组细胞凋亡情况。结果:与对照组比较,损伤组NRK-52E细胞肿大,胞核不规则样改变,凋亡细胞增多,凋亡相关蛋白Bax上调而Bcl-2下调;处理组NRK-52E细胞肿大较损伤组缓解,胞核形态改善,凋亡细胞减少,Bax下调且Bcl-2上调。结论:大鼠骨髓MSC来源的外切体对顺铂诱导的NRK-52E细胞凋亡有一定的抑制作用。Objective: To investigate the effect of exosome derived from rat bone marrow mesenchymal stem cells on the apoptosis of cisplatin-induced renal proximal tubular epithelial cells. Methods: Primitive rat bone marrow mesenchymal stem cells were separated and cultured in vitro from which exosome were extracted and proofed. The injury of NRK-52 E cells was induced by 5 μmol / L cisplatin for 6 h and cultured for another 48 h using normal DMEM. 100 μg / m L exosome contained DMEM was used in restoration group for the 48 h after the cisplatin treatment,and normal NRK-52 E cells with no treatment were used as control. TUNEL-FITC,flow cytometry and immunoblotting were used to detect the apoptosis in each group.Results: Compared with the control group,NRK-52 E cells in cisplatin group grew into swollen ones with nucleus changed irregularly,the number of apoptotic cells was increased; the expression of apoptosis associated protein Bax was upregulated while Bcl-2 downregulated. However,in the exosome group,the phenomenon of swell and irregular nucleus in NRK-52 E cells was alleviated. The number of apoptotic cells was decreased,with decreased expression of Bax and increased expression of Bcl-2. Conclusion: Exosome derived from rat bone marrow mesenchymal stem cells could suppress the apoptosis induced by cisplatin in NRK-52 E cells.

关 键 词：外切体 顺铂 肾小管上皮细胞 损伤 凋亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]