微小RNA-193a调控Wilms瘤基因1促进小鼠糖尿病肾病足细胞凋亡

高飞; 王泽泽; 杨冰; 檀金川

doi:10.16098/j.issn.0529-1356.2021.05.009

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解剖学报 ›› 2021, Vol. 52 ›› Issue (5) : 728-736. DOI: 10.16098/j.issn.0529-1356.2021.05.009

细胞和分子生物学

微小RNA-193a调控Wilms瘤基因1促进小鼠糖尿病肾病足细胞凋亡

高飞¹张欣欣¹ 杨冰¹ 陈素枝² 杨凤文² 檀淼³ 檀金川^2*

作者信息 +

MicroRNA-193a regulating Wilms’tumor gene 1 to promote mouse podocyte apoptosis in diabetic nephropathy#br#

GAO Fei¹ ZHANG Xin-xin¹ YANG Bing¹ CHEN Su-zhi² YANG Feng-wen² TAN Miao³ TAN Jin-chuan^2*#br#

Author information +

文章历史 +

摘要

目的探讨微小RNA（miR）-193a对糖尿病肾病（DN）足细胞凋亡的影响及作用机制。方法通过体外高糖培养足细胞和体内小鼠腹腔注射链脲佐菌素(STZ)复制DN模型，将细胞或60只小鼠随机分为空白对照组、模型对照组、miR-193a抑制阴性对照组、miR-193a抑制组、miR-193a过表达阴性对照组和miR-193a过表达组。使用流式细胞术、免疫组织化学、免疫荧光、TUNEL、Real-time PCR和Western blotting检测DN小鼠和小鼠足细胞凋亡情况。结果 DN小鼠和高糖诱导的小鼠足细胞中Nephrin、Podocin表达减弱，细胞凋亡率显著升高，miR-193a高表达，小鼠肾脏和足细胞中cleaved-Caspase-3和Bax蛋白水平显著升高，Bcl-2蛋白水平显著下降，而miR-193a 抑制剂(inhibitor)可改善这一过程。DN小鼠和高糖培养的小鼠足细胞中Wilms瘤基因1（WT1） mRNA和蛋白表达水平显著降低，miR-193a inhibitor干预后WT1蛋白表达显著升高。上调WT1可降低miR-193a对高糖诱导的小鼠足细胞凋亡的影响。双荧光素酶报告实验证实了miR-193a和WT1之间的靶向关系。
结论 MiR-193a下调WT1的表达，促进DN足细胞凋亡。

Abstract

Objective To explore the effect of microRNA(miR)-193a on the apoptosis of mouse podocytes in diabetic nephropathy (DN) and its mechanism. Methods The DN model was replicated by culturing podocytes with high glucose in vitro and intraperitoneal injection of streptozocin (STZ) in mice in vivo The cells or 60 mice were randomly divided into normal control(NC) group, model control group, and miR-NC inhibitor group, miR-193a inhibitor group, miR-NC mimic group and miR-193a mimic group. Flow cytometry, immunohistochemistry, TUNEL, Real-time PCR, Western blotting were used to examine the apoptosis of DN mice and mouse podocytes. Results The expression of Nephrin and Podocin in podocytes was weakened in DN mice and renal podocytes induced by high glucose, the apoptotic rate increased significantly, miR-193a was highly expressed, the levels of cleaved-Caspase-3 and Bax protein increased significantly, the level of Bcl-2 protein decreased significantly, and miR-193a inhibitor could improve this process. Wilms’tumor gene 1(WT1) mRNA and protein expression levels were significantly reduced in DN mice and podocytes cultured with high glucose. WT1 protein expression increased significantly after miR-193a inhibitor intervention, and WT1 protein expression was significantly reduced after miR-193a mimic transfection. Up-regulating WT1 could reduce the effect of miR-193a on the apoptosis of mouse podocytes induced by high glucose. The dual luciferase reporter experiment confirmed the targeting relationship between miR-193a and WT1. Conclusion MiR-193a down-regulates the expression of WT1 and promotes apoptosis of DN podocytes.

导出引用

高飞张欣欣杨冰陈素枝杨凤文檀淼檀金川. 微小RNA-193a调控Wilms瘤基因1促进小鼠糖尿病肾病足细胞凋亡[J]. 解剖学报. 2021, 52(5): 728-736 https://doi.org/10.16098/j.issn.0529-1356.2021.05.009

GAO Fei ZHANG Xin-xin YANG Bing CHEN Su-zhi YANG Feng-wen TAN Miao TAN Jin-chuan. MicroRNA-193a regulating Wilms’tumor gene 1 to promote mouse podocyte apoptosis in diabetic nephropathy#br#[J]. Acta Anatomica Sinica. 2021, 52(5): 728-736 https://doi.org/10.16098/j.issn.0529-1356.2021.05.009

中图分类号： R587.2

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