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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。
AIM: To investigate the mechanism of miR-153 in regulating high glucose-induced renal tubular epithelial cells (HK2) -mesenchymal transition (EMT) and to provide new ideas for studying the mechanism of interstitial fibrosis in diabetic nephropathy (DN). METHODS: Bioinformatics methods were used to predict the candidate miRNAs that regulate Snail. All candidate miRNAs were verified in db / db mice and pearson correlation analysis was performed with Snail respectively. The most significant miRNAs were found to be negatively correlated with Snail. HK2 cells induced by high glucose were used as vectors, and the miRNA mimics were transfected for 72 h. Real-time PCR and western blot were used to observe the changes of miRNAs, Snail and E-cadherin. Results: Seven candidate miRNAs that regulate Snail were miR-153, miR-30e, miR-30d, miR-30b, miR-384-5p, miR-30a and miR- The expression of miR-153, miR-30d and miR-30e were significantly downregulated in renal cortex of db / db mice (P <0.05). High glucose induced HK2 cells down-regulated miR-153, tubular epithelial cell marker E-cadherin expression decreased; over-expression of miR-153 inhibited HK2 cells express Snail, while the E-cadherin level increased. Conclusion: MiR-153 may be involved in the regulation of EMT induced by high glucose in HK2 cells through the target gene Snail.