miR-30e*在顺铂诱导的肾近端小管上皮细胞凋亡和线粒体损伤中的作用  

Role of mi R-30e* in cisplatin induced renal proximal tubular cells apoptosis and mitochondrial injury

作　　者：杨玲云[1] 郭燕[1] 倪佳佳[1] 王荣[1] 林加娟[1] 丁桂霞[1] 黄松明[1] 张爱华[1] 

机构地区：[1]南京医科大学附属南京儿童医院肾脏科,江苏南京210008

出　　处：《南京医科大学学报（自然科学版）》2015年第8期1060-1065,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助(81270797)

摘　　要：目的:观察顺铂诱导肾近端小管细胞损伤过程中mi R-30e*的表达变化,并探讨其在顺铂诱导的小管细胞凋亡和线粒体损伤中的作用。方法:实时定量PCR(q RT-PCR)检测顺铂处理后的小管细胞mi R-30e*的表达水平;使用重组慢病毒载体感染细胞,稳定高表达或低表达mi R-30e*;使用流式细胞仪检测细胞凋亡,观察mi R-30e*在顺铂诱导的小管细胞凋亡中的作用;q RT-PCR检测线粒体DNA(mt DNA)拷贝数,JC-1检测线粒体膜电位变化,探究mi R-30e*在顺铂引起的线粒体损伤中的作用。结果:1顺铂处理肾小管上皮细胞后,mi R-30e*的表达呈时间依赖性和剂量依赖性下调;2使用q RT-PCR检测重组慢病毒载体制备的稳定高表达或低表达mi R-30e*的小管细胞株中mi R-30e*的表达,过表达组增高了7倍,敲低表达组降低了近50%;3加入顺铂后,与对照组相比小管细胞凋亡增加,过表达mi R-30e*对凋亡起保护作用,敲低mi R-30e*则加重凋亡;4通过检测mt DNA的拷贝数发现顺铂诱导的线粒体损伤在48 h时有意义,迟于mi R-30e*表达的下调,线粒体膜电位检测JC-1提示过表达mi R-30e*对线粒体有保护作用。结论:顺铂处理小管细胞可降低其mi R-30e*的表达;体外过表达mi R-30e*对顺铂引起的小管细胞的凋亡和线粒体损伤具有保护作用,敲低mi R-30e*会加重顺铂诱导的肾近端小管细胞凋亡和线粒体损伤。Objective:To investigate the expression of mi R-30e*and the role of mi R-30e*in cisplatin induced mouse proximal tubular cells(m PTCs) apoptosis and mitochondrial injury. Methods:Expression of mi R-30e*in m PTCs treated with cisplatin were determined by quantitative real-time PCR(q RT-PCR). To stably over expression or knock down mi R-30e*, recombinant lentiviral expressing vectors were used for transfection of m PTCs. Flow cytometry was performed to analyze the apoptosis of m PTCs after transfection and treated with or without cisplatin. Mitochondrial DNA(mt DNA) copy numbers were determined by q RT-PCR and mitochondrial membrane potential was examined by JC-1 staining. Results:Cisplatin reduced the expression levels of mi R-30e*in a dose and time-dependent manner. MPTCs transfected with over expression mi R-30e*recombinant lentiviral expressing vector gained an increased expression of mi R-30e*for seven fold. While knock down of mi R-30e*decreased the expression of mi R-30e*for 50%compared with vehicle. The apoptosis of m PTCs increased when treated with cisplatin,over expression mi R-30e*reduced cisplatin induced apoptosis and knock down of mi R-30e*facilitated cisplatin induced apoptosis. mt DNA significantly decreased after cisplatin treatment for 48 h,later than the decrease expression of mi R-30e*. JC-1 staining revealed ectopic mi R-30e*can protect mitochondrial membrane from cisplatin induced injury. Conclusion:Cisplatin reduced the expression levels of mi R-30e*in vitro. Ectopic mi R-30e*can protect m PTCs from apoptosis and mitochondrial membrane potential changes induced by cisplatin.

关 键 词：miR-30e* 顺铂 肾近端小管上皮细胞 线粒体损伤 

分 类 号：R692[医药卫生—泌尿科学]