miR-30b/Snail调控糖尿病肾病肾小管上皮细胞EMT  

The Role of miR-30b/Snail in EMT of DN Renal Tubular Epithelial Cells

作　　者：王彦哲[1] 王筱霞[1,2] 汪年松[1] 

机构地区：[1]上海交通大学附属第六人民医院肾脏内科,上海200233 [2]上海交通大学医学院附属同仁医院肾脏内科,上海200336

出　　处：《中国中西医结合肾病杂志》2017年第4期288-291,I0001,I0002,共6页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然基金面上项目(No.81270814;81270824);上海市国际科技合作基金项目(No.11410708500)

摘　　要：目的:探讨miR-30b/Snail调控高糖诱导人源肾小管上皮细胞(HK2)-间充质转化(EMT)的可能机制,为糖尿病肾病(DN)间质纤维化的防治提供新的实验依据及思路。方法:通过生物信息学预测调控Snail的候选miRNAs;在db/db小鼠肾组织中验证所有候选miRNAs的表达并分别与Snail做pearson相关性分析,筛选出与Snail负相关最显著的miRNA;以高糖诱导的HK2细胞为载体,转染该miRNA mimics 72 h后,采用real-time PCR、western blot方法,观察该miRNA、Snail以及EMT相关蛋白的表达改变。结果:调控Snail的候选miRNAs共6个,分别是miR-30b-5p、miR-25-3p、miR-199a-5p、miR-128-3p、miR-22-3p、miR-27-3p,其中miR-30b与Snail存在明显负相关,r2=0.775 49;高糖诱导HK2细胞miR-30b表达下调,肾小管上皮细胞标志蛋白E-cadherin表达减少;过表达miR-30b-5p可抑制HK2细胞Snail表达,同时上调E-cadherin表达,而Vimentin、α-SMA表达下调。结论:miR-30b/Snail参与调控高糖引起的肾小管上皮细胞EMT。Objective： To investigate mechanism by miR - 30b/Snail regulate high glucose - induced the epithelial - mesenchymal transition （EMT） in human renal proximal tubule cell （HK2）, providing new evidence and ideas in prevention and treatment of DN renal interstitial fibrosis. Methods ： Using bioinformatics to predict the candidates of miRNAs which potentially target Snail, then we confirmed the expressions of all of miRNA candidates in renal cortex of db/db mice. Pearson relevant analysis was performed between miRNAs and Snail respectively. We selected the miRNA which showing the most significantly negative relevance with Snail. This miRNA and Snail mRNA were detected in HK2 cells exposure to high glucose condition by using real -time PCR. The protein levels of Snail, E - cadherin and EMT related proteins in HK2 cells were measured 72 hours after transfeetion of selected miRNA mimics. Results：We have found 6 miRNAs candidates potentially targeting Snail through bioinformatics, which were miR -30b -5p, miR - 25 - 3p, miR - 199a - 5p, miR - 128 - 3p, miR - 22 - 3p, miR - 27 - 3p. Among these candidates, miR - 30b was significantly negative relevance with Snail, R^2 =0.77549. We found down - regulation of the expression of miR - 30b and E - cadherin in high glucose - induced HK2 cells. Overexpression of miR - 30b decreased the levels of Snail, whereas E - eadherin increased in cultured HK2 cells instead, while down- regulate the expression of Vimentin andα- SMA. Conclusion：miR- 30b/Snail is involved in the regulation of high glucose - induced renal tubule epithelial cell EMT.

关 键 词：近端肾小管上皮细胞 上皮细胞-间充质转化(EMT) miR-30b SNAIL 生物信息学 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]