下调miR-153促进高糖诱导的肾小管上皮细胞-间充质转化

Down-regulation of miR-153 Contributes to High Glucose- mediated Renal Tubule EMT

目的:探讨miR-153调控高糖诱导肾小管上皮细胞(HK2)-间充质转化(EMT)的机制,为研究糖尿病肾病(DN)间质纤维化机制提供新思路。方法:采用生物信息学方法预测调控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miRNAs、Snail以及E-cadherin的表达改变。结果:调控Snail的候选miRNAs共7个,分别是:miR-153、miR-30e、miR-30d、miR-30b、miR-384-5p、miR-30a、miR-30c;与db/m相比,miR-153、miR-30d及miR-30e在db/db小鼠肾皮质中表达显著下调(P<0.05),其中miR-153与Snail表达水平负相关最显著(r=-0.501 56);高糖诱导HK2细胞下调miR-153,肾小管上皮细胞标志蛋白E-cadherin表达减少;过表达miR-153抑制HK2细胞表达Snail,而E-cadherin水平增加。结论:MiR-153可能通过靶基因Snail参与调控高糖诱导的HK2细胞EMT。

Objective：To investigate mechanism by which miR-153 regulates high glucose-induced the epithelial-mesenchymal transition（EMT） in human renal proximal tubule cell（HK2）,providing a novel insight into interstitial fibrosis in the process of DN.Methods：We used bioinformatics to predict the candidates of miRNAs potentially targeting Snail,followed by confirming the expressions of all of miRNA candidates in renal cortex of db/db mice;Pearson relavant analysis was performed between miRNAs and Snail respectively.We further focus on miRNA showing the most significantly negative relevance with Snail.The selected 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 in HK2 cells were measured 72 hours after transfection of selected miRNA mimics.Results：A total of 7 miRNA candidates potentially targeting Snail were screened by bioinformatics,which were miR-153,miR-30e,miR-30d,miR-30b,miR-384-5p,miR-30a,miR-30c.Among these candidates,miR-153、miR-30d and miR-30e down-regulated remarkably in renal cortex in db/db mice compare with db/m controls（P0.05）.Pearson analysis indicated only miR-153 showing the most significantly negative relevance with Snail（r=-0.501 56）.The similar results were found in high glucose-induced HK2 cells.Overexpression of miR-153 decreased the levels of Snail,whereas E-cadherin increased in cultured HK2 cells instead.Conclusion：The current study reveals that miR-153 might involve in the regulation of high glucose-mediated tubule epithelial cell EMT via targeting Snail.