miR-660-5p调控P53对高糖诱导的肾小球系膜细胞损伤的影响  

Effect of miR-660-5p on High Glucose-induced Glomerular Mesangial Cell Injury by Regulation of P53

作　　者：谷梁 马小溪 金志敏 GU Liang;MA Xiaoxi;JIN Zhimin(Department of Internal Medicine,Hebei Provincial Ophthalmology Hospital,Xingtai,Hebei,054001,China)

机构地区：[1]河北省眼科医院内科,河北省邢台市054001

出　　处：《医学分子生物学杂志》2024年第6期557-561,共5页Journal of Medical Molecular Biology

基　　金：2021年度河北省医学科学研究课题计划(No.20210540)。

摘　　要：目的探索miR-660-5p调控P53对高糖诱导的肾小球系膜细胞损伤的影响。方法首先将肾小球系膜细胞SV40-MES-13细胞分为2组:对照组和高糖组,用于分析高糖对SV40-MES-13细胞活性及miR-660-5p/P53表达的影响。然后将高糖处理过后的SV40-MES-13细胞分为4组:miR NC组、miR-660-5p mimic组、si NC组与si P53组。实时荧光定量PCR实验检测SV40-MES-13细胞中miR-660-5p和P53的表达水平;噻唑蓝试剂法(MTT)分析SV40-MES-13细胞的生长能力;原位末端标记法(TUNEL)以及流式细胞术分析SV40-MES-13细胞的凋亡率;Targetscan检索以及荧光素酶报告基因实验分析SV40-MES-13细胞中miR-660-5p与P53的靶向作用关系;蛋白质印迹分析P53蛋白的表达。结果与对照组比较,高糖处理后导致SV40-MES-13细胞增殖活性降低,凋亡率增加,miR-660-5p表达降低而P53表达增加。与miR-660-5p NC组比较,miR-660-5p mimic处理后的SV40-MES-13细胞增殖活性增加,凋亡率降低。与si NC组比较,si P53处理后的SV40-MES-13细胞增殖活性增加,凋亡率降低。miR-660-5p与P53存在结合域,且能抑制P53表达。结论高糖处理肾小球系膜细胞后,miR-660-5p表达水平降低,P53表达水平增加,上调肾小球系膜细胞的miR-660-5p的表达后,高糖处理后的肾小球系膜细胞增殖活性增加,凋亡率下降,并且miR-660-5p的这一作用与抑制P53的表达相关。Objective To investigate the effect of miR-660-5p on high glucose-induced glomerular mesangial cell injury by regulating P53.Methods SV40-MES-13 cells were divided into two groups:control group and high glucose group,to analyze the effect of high glucose on cell viability and expression of miR-660-5p/P53.Then SV40-MES-13 cells treated with high glucose were divided into four groups:miR NC group,miR-660-5p mimic group,siNC group and siP53 group.The expression levels of miR-660-5p and P53 in SV40-MES-13 cells were detected by real-time fluorescence quantitative PCR.The growth ability of SV40-MES-13 cells was analyzed by methyl thiazolyl tetrazolium(MTT)assay.The apoptosis rate of SV40-MES-13 cells was analyzed by TUNEL and flow cytometry.Targetscan and luciferase reporter assay were used to analyze the targeting relationship between miR-660-5p and P53 in SV40-MES-13 cells.The expression level of P53 protein was analyzed by Western blotting.Results Compared with cells in the control group,SV40-MES-13 cells treated with high glucose showed decreased proliferation activity,increased apoptosis rate,decreased expression level of miR-660-5p and increased expression level of P53.The proliferation activity of SV40-MES-13 cells treated with miR-660-5p mimic was increased,and the apoptosis rate was decreased when compared with those in the miR-660-5p NC group.The proliferation activity of SV40-MES-13 cells treated with siP53 was increased,and the apoptosis rate was decreased when compared with those in the siNC group.miR-660-5p could bind with P53 and inhibit the expression of P53.Conclusion The expression level of miR-660-5p is decreased and the expression level of P53 is increased in high glucose-treated mesangial cells.Up-regulation of miR-660-5p expression in high glucose-treated mesangial cells increases the proliferation activity and decreases the apoptosis rate,and the effect of miR-660-5p is related to the inhibition of P53 expression.

关 键 词：miR-660-5p P53 肾小球系膜细胞 SV40-MES-13 糖尿病肾病 凋亡 增殖 

分 类 号：R692.9[医药卫生—泌尿科学]