[1]李梦岚,谭琴,徐秀.miRNA-192靶向调控WT1在高糖诱导 足细胞EMT中的作用[J].国际内分泌代谢杂志,2020,40(06):361-366.[doi:10.3760/cma.j.cn121383-20200310-03025]
　Li Menglan,Tan Qin,Xu Xiu.The role of miRNA-192 targeted regulation of WT1 in high glucose induced EMT in podocytes[J].International Journal of Endocrinology and Metabolism,2020,40(06):361-366.[doi:10.3760/cma.j.cn121383-20200310-03025]
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miRNA-192靶向调控WT1在高糖诱导 足细胞EMT中的作用()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
40
期数:
2020年06期
页码:
361-366
栏目:
论著
出版日期:
2020-11-20

文章信息/Info

Title:
The role of miRNA-192 targeted regulation of WT1 in high glucose induced EMT in podocytes
作者:
李梦岚1; 谭琴1; 徐秀2
1武汉市中心医院内分泌科 430000; 2华中科技大学同济医学院附属同济医院肾内科,武汉 430000
Author(s):
Li Menglan1;  Tan Qin1;  Xu Xiu2
1Department of Endocrinology, The Central Hospital of Wuhan, Wuhan 430000, China; 2Department of Nephrology, Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430000, China
关键词:
miRNA-192;  Wilms肿瘤蛋白1;  糖尿病肾病;  足细胞;  上皮-间充质转化
Keywords:
miRNA-192;  Wilms' tumor 1 protein;  Diabetic nephropathy;  Podocytes;  Epithelial-mesenchymal transition
DOI:
10.3760/cma.j.cn121383-20200310-03025
文献标志码:
A
摘要:
目的 探讨miRNA-192靶向调控Wilms肿瘤蛋白1(WT1)在高糖诱导足细胞上皮-间充质转化(EMT)中的作用。方法 以人肾小球足细胞为研究对象,将细胞分为4组:空白对照组、高糖组、阴性对照组、miRNA-192抑制组。采用Western印迹检测各组足细胞表面标志蛋白肾病蛋白(nephrin)、WT1、转化生长因子(TGF)-β1、α-平滑肌肌动蛋白(α-SMA)的蛋白表达水平; qRT-PCR检测各组细胞miRNA-192、WT1、TGF-β1、α-SMA mRNA表达水平; 采用双荧光素酶靶标实验验证miRNA-192与WT1基因的靶向关系。结果 与空白对照组相比,高糖组和阴性对照组nephrin的蛋白表达水平显著降低,差异有统计学意义(F=444.404,P<0.05); miRNA-192相对表达量升高而WT1蛋白及mRNA表达水平均降低(F=184.216、243.543、107.898,P均<0.05); 双荧光素酶报告基因实验分析结果显示,WT1是miRNA-192的靶基因; 与空白对照组相比,高糖组和阴性对照组TGF-β1、α-SMA蛋白及mRNA表达水平均升高(FTGF-β1蛋白=69.014,P<0.05; Fα-SMA蛋白=87.644,P<0.05; FTGF-β1 mRNA=147.714,Fα-SMA mRNA=247.584,P<0.05)。与阴性对照组相比,miRNA-192抑制组nephrin的蛋白表达水平升高(t=16.519,P<0.05),miRNA-192相对表达量降低、WT1蛋白及mRNA表达水平升高(tmiRNA-192=10.533,tWT1=17.088、8.186,P均<0.05); TGF-β1、α-SMA蛋白及mRNA蛋白表达水平均降低(tTGF-β1蛋白=6.114,P<0.05; tα-SMA蛋白=7.941,P<0.05; tTGF-β1 mRNA=8.239,tα-SMA mRNA=8.481,P均<0.05)。结论 miRNA-192靶向调控WT1促进糖尿病肾病的发展,其作用机制可能与足细胞EMT有关; 抑制miRNA-192的表达可减轻高糖诱导的足细胞EMT。
Abstract:
Objective To investigate the role of miRNA-192 in the regulation of Wilms' tumor 1 protein(WT1)induced epithelial-mesenchymal transition(EMT)in podocytes.Methods Human glomerular podocytes were divided into four groups: blank control group, high glucose group, negative control group, miRNA-192 inhibition group. Western blotting was used to detect the expression level of the surface marker proteins nephrin, WT1, transforming growth factor-β1(TGF-β1)and α-smooth muscle actin(α-SMA)in podocytes of each group. The expression of miRNA-192 and WT1 mRNA in each group were detected by qRT-PCR. The targeting relationship between miRNA-192, WT1, TGF-β1 and α-SMA gene was verified by double luciferase target experiment.Results Compared with blank control group, the expression level of nephrin in high glucose group and negative control group was significantly lower(F=444.404, P<0.05). The relative expression of miRNA-192 increased while that of WT1 protein and mRNA decreased(F=184.216, 243.543, 107.898, all P<0.05). WT1 was the target gene of miRNA-192 according to the analysis of double luciferase reporter gene. Compared with blank control group, the expression levels of TGF-β1, α-SMA protein and mRNA in high glucose group and negative control group were increased(FTGF-β1 protein=69.014, P<0.05; Fα-SMA protein =87.644, P<0.05; FTGF-β1 mRNA=147.714, Fα-SMA mRNA=247.584, all P<0.05). Compared with negative control group, the expression level of nephrin in miRNA-192 inhibition group was increased(t=16.519, P<0.05), the relative expression of miRNA-192 was decreased, and the relative expression of WT1 protein and mRNA were increased(tmiRNA-192=10.533, tWT1=17.088, 8.186, all P<0.05). The expression levels of TGF-β1, α-SMA protein and mRNA protein were decreased(tTGF-β1 protein=6.114, P<0.05; tα-SMA protein=7.941, P<0.05; tTGF-β1 mRNA=8.239, tα-SMA mRNA=8.481, P<0.05).Conclusion miRNA-192 can promote the development of diabetic nephrophy by targetting WT1. Inhibition of miRNA-192 expression can reduce the EMT induced by high glucose in podocytes.

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备注/Memo

备注/Memo:
通信作者:李梦岚,Email:limenglan56@163.com
更新日期/Last Update: 2020-11-20