MiR-30c-2-3p对足细胞活力及Nephrin和Podocin蛋白表达的影响

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摘要目的探讨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 < 0.01），阴性质粒+PAN组较PAN组差异无统计学意义（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或P < 0.01）。②qRT-PCR显示PAN组MiR-30c-2-3p的表达较空白组明显下降，差异有高度统计学意义（P < 0.01）；阴性质粒+PAN组较PAN组表达差异无统计学意义（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或P < 0.01）。③Western blot提示PAN组Nephrin和Podocin蛋白的表达较空白组明显下降，差异有高度统计学意义（P < 0.01）；与PAN组相比，阴性质粒+PAN组两者蛋白的表达差异无统计学意义（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。

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	陈琪1 闵晶晶2 王霄一1▲

关键词 ： MiR-30c-2-3p, 足细胞, 嘌呤霉素, 细胞活性, Nephrin, Podocin

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 < 0.01). There was no significant difference between the negative plasmid+PAN group and the PAN group (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 or P < 0.01). ②The expression of MiR-30c-2-3p in the PAN group was significantly lower than that in the blank group, the differences were statistically significant (P < 0.01). There was no significant difference between the negative plasmid group and the PAN group (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 significant (P < 0.05 or P < 0.01). ③The expression of Nephrin and Podocin in the PAN group were significantly lower than those in the blank group, the differences were statistically significant (P < 0.01). There was no significant difference between the negative plasmid+PAN group and the PAN group (P > 0.05). Compared with the PAN group and the negative plasmid+PAN group, the expression of Nephrin and Podocin in the positive plasmid+PAN group increased, the differences were statistically significant (P < 0.05). Conclusion The PAN intervention inhibits the podocyte activity, MiR-30c-2-3p content, and the Nephrin and Podocin expression, but the exogenous expression of MiR-30c-2-3p can improve the viability of podocytes and increase the expression of Nephrin and Podocin protein.

Key words： MiR-30c-2-3p Podocyte Puromycin Viability Nephrin Podocin

基金资助:浙江省医药卫生科技项目（2015DTA017）；
浙江省湖州市科技局公益性应用研究项目（2016GYB25）。

通讯作者: ▲通讯作者

引用本文:

陈琪1 闵晶晶2 王霄一1▲. MiR-30c-2-3p对足细胞活力及Nephrin和Podocin蛋白表达的影响[J]. 中国医药导报, 2017, 14(28): 17-20,30.
CHEN Qi1 MIN Jingjing2 WANG Xiaoyi1▲. Effects of MiR-30c-2-3p on podocyte viability and expression of Nephrin and Podocin. 中国医药导报, 2017, 14(28): 17-20,30.

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