MiR―30c―2―3p对足细胞活力及Nephrin和Podocin蛋白表达影响.docVIP

MiR―30c―2―3p对足细胞活力及Nephrin和Podocin蛋白表达影响 　　[摘要] 目的 探?MiR-30c-2-3p对足细胞活力及Nephrin和Podocin蛋白的表达影响。 方法 采用嘌呤霉素（PAN）诱导的足细胞体外模型，将足细胞分成4组，即空白组、PAN组、阴性质粒+PAN组、阳性质粒+PAN组。通过LipofectamineTM 2000瞬时转染MiR-30c-2-3p mimic。通过Cell Counting Kit-8（CCK-8）法检测细胞活力、实时荧光定量PCR（qRT-PCR）检测细胞中MiR-30c-2-3p表达、Western blot检测Nephrin和Podocin蛋白的表达。 结果 ①CCK-8结果提示PAN组足细胞活力明显低于空白组，差异有高度统计学意义（P 0.05），阳性质粒+PAN组足细胞活力与PAN组和阴性质粒+PAN组相比均有所提高，差异有统计学意义（P 0.05），而阳性质粒+PAN组MiR-30c-2-3p的表达较PAN组和阴性质粒+PAN组均提高，差异有统计学意义（P 0.05），而阳性质粒+PAN组Nephrin和Podocin蛋白的表达较单纯PAN组和阴性质粒+PAN组均增加，差异有统计学意义（P 0.05）。 结论 PAN对足细胞的活力，MiR-30c-2-3p含量及Nephrin和Podocin蛋白的表达有抑制作用，而通过外源性增加MiR-30c-2-3p表达后可提高足细胞的活力，同时可增加Nephrin和Podocin蛋白的表达。 　　[关键词] MiR-30c-2-3p；足细胞；嘌呤霉素；细胞活性；Nephrin；Podocin 　　[中图分类号] R692 [文献标识码] A [文章编号] 1673-7210（2017）10（a）-0017-05 　　[Abstract] Objective To investigate the effect of MiR-30c-2-3p on podocyte viability and Nephrin and Podocin protein expression. Methods The model of mice podocyte injury was induced by puromycin aminonucleoside （PAN） in vitro， the podocytes were divided into four groups： blank group， PAN group， negative plasmids+PAN group， positive plasmids+PAN group. The MiR-30c-2-3p mimics were transiently transfected with LipofectamineTM 2000， the cell viability was detected by Cell Counting Kit-8 （CCK-8）， the expression of MiR-30c-2-3p was detected by real-time quantitative PCR （qRT-PCR）， the expression of Nephrin and Podocin protein was detected by Western blot. Results ①CCK-8 showed that the viability of PAN group was significantly lower than that of the blank group， the differences were statistically significant （P 0.05）. The viability of positive plasmid+PAN group was significantly increased than that of the PAN group and the negative plasmid+PAN group， the differences were statistically significant （P 0.05）. The content of MiR-30c-2-3p in the positive plasmid+PAN group was significantly higher than that of the PAN group and the negative plasmid+PAN group， the differences were statistically significan