论文部分内容阅读
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 observe the expression of mi R-30e * in cisplatin-induced renal proximal tubular cell injury and to explore its role in cisplatin-induced apoptosis of tubular cells and mitochondrial damage. Methods: Real-time quantitative PCR (q RT-PCR) was used to detect the expression of mi R-30e * in cisplatin-treated tubule cells. The recombinant lentiviral vector was used to infect cells to stably overexpress or downregulate mi R-30e * The effect of mi R-30e * on cisplatin-induced apoptosis was observed by flow cytometry. The copy number of mitochondrial DNA (mt DNA) was detected by q RT-PCR, the mitochondrial membrane potential was detected by JC- Explore the role of mi R-30e * in mitochondrial-induced mitochondrial damage. Results: 1 The expression of mi R-30e * was down-regulated in cisplatin-treated renal tubular epithelial cells in a time-and dose-dependent manner.2 The stable or highly expressed mi R -30e * expression in the tubular cell line mi R-30e *, over-expression group increased 7-fold, knockdown expression group decreased by nearly 50%; 3 after adding cisplatin compared with the control group increased tubular apoptosis , Overexpression of mi R-30e * protected against apoptosis, knockdown of mi R-30e * increased apoptosis.4 Detection of cisplatin-induced mitochondrial damage was significant at 48 h by detection of mt DNA copy number Downregulation of mi R-30e * expression, detection of mitochondrial membrane potential JC-1 suggests that overexpression of mi R-30e * has a protective effect on mitochondria. CONCLUSION: Cisplatin can reduce the expression of mi R-30e * in tubular cells. In vitro overexpression of mi R-30e * has a protective effect on cisplatin-induced tubular cell apoptosis and mitochondrial damage, knocking down mi R-30e * Will increase cisplatin-induced renal proximal tubule cell apoptosis and mitochondrial damage.