[1]乐颖,蒋科威,薛萌,等.microRNA-31对高糖条件下人足细胞上皮-间充质转分化的影响[J].国际内分泌代谢杂志,2022,42(05):354-359.[doi:10.3760/cma.j.cn121383-20210426-04077]
　Le Ying,Jiang Kewei,Xue Meng,et al.Effects of microRNA-31 on high-glucose induced epithelial-mesenchymal transition of podocytes[J].International Journal of Endocrinology and Metabolism,2022,42(05):354-359.[doi:10.3760/cma.j.cn121383-20210426-04077]
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microRNA-31对高糖条件下人足细胞上皮-间充质转分化的影响()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
42
期数:
2022年05期
页码:
354-359
栏目:
论著
出版日期:
2022-09-20

文章信息/Info

Title:
Effects of microRNA-31 on high-glucose induced epithelial-mesenchymal transition of podocytes
作者:
乐颖1; 蒋科威2; 薛萌1; 张秀珍1; 袁凤易1
1深圳市人民医院、暨南大学第二临床医学院、南方科技大学第一附属医院内分泌科 518020; 2深圳市人民医院、暨南大学第二临床医学院、南方科技大学第一附属医院老年病科 518020
Author(s):
Le Ying1;  Jiang Kewei2;  Xue Meng1;  Zhang Xiuzhen1;  Yuan Fengyi1.
1Department of Endocrinology and Metabolism, Shenzhen People's Hospital, Second Affiliated Hospital of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology, Shenzhen 518020, China; 2Department of Geriatrics, Shenzhen People's Hospital, Second Affiliated Hospital of Jinan University, the First Affiliated Hospital of Southern University of Science and Technology, Shenzhen 518020, China
关键词:
miR-31;  低氧诱导因子-1抑制剂;  糖尿病肾脏病;  足细胞;  上皮-间充质转分化
Keywords:
miR-31;  FIH-1;  Diabetic kidney disease;  Podocyte;  Epithelial-mesenchymal transition
DOI:
10.3760/cma.j.cn121383-20210426-04077
摘要:
目的 探讨microRNA-31(miR-31)在高糖诱导足细胞上皮-间充质转分化(EMT)中的机制。方法 将体外培养的人肾小球足细胞按不同糖浓度分为低糖组(LG)、高渗组(HM)和高糖组(HG); 按是否转染过表达miR-31(miR-31 mimics)分为miR-31过表达组(miR-31m组)、阴性对照组(miR-NC组)和脂质体组(Mock组); 按照是否转染沉默低氧诱导因子-1抑制剂(FIH-1)及沉默miR-31(miR-31 inhibitor)分为FIH-1沉默组(si-FIH-1组)、miR-31沉默组(miR-31i组)、FIH-1沉默+miR-31沉默组(si-FIH-1+miR-31i组)及阴性对照组(NC组)。采用实时聚合酶链式反应(qRT-PCR)和Western blot检测各组细胞低氧诱导因子-1抑制剂(FIH-1)、转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)的mRNA和蛋白表达水平; 采用双荧光素酶靶标实验验证miR-31与FIH-1基因的靶向关系。结果 与LG和HM组比较,HG组miR-31表达水平显著升高(F=146.8,P<0.01),FIH-1蛋白表达水平明显下降(F=54.23,P<0.01),而TGF-β1及α-SMA蛋白表达水平均显著升高(F=360.6,P<0.01; F=193.7,P<0.01)。双荧光素酶报告基因实验分析结果显示,FIH-1是miR-31的靶基因。si-FIH-1与miR-31i共同转染时,可以恢复si-FIH-1或miR-31i单独转染导致的FIH-1、TGF-β1、α-SMA的mRNA及蛋白表达变化。结论 miR-31靶向调控FIH-1促进足细胞EMT,抑制miR-31的表达可减轻高糖诱导的足细胞EMT。
Abstract:
Objective To investigate the mechanism of microRNA-31(miR-31)in high-glucose induced epithelial-mesenchymal transition(EMT)of podocytes.Methods According to different sugar concentrations, human glomerular podocytes cultured in vitro were divided into low glucose group(LG), hyperosmolar group(HM)and high glucose group(HG); according to whether transfected miR-31 mimics, podocytes were divided into miR-31m group, negative miR-NC group and Mock group; according to whether transfected si-FIH-1 and miR-31i, podocytes were divided into si-FIH-1 group, miR-31i group, si-FIH-1+miR-31i group and NC group. Western blot was used to detect the protein expression levels of FIH-1, TGF-β1, and α-SMA in podocytes of each group; qRT-PCR was used to detect the mRNA expression levels of miR-31, FIH-1, TGF-β1, and α-SMA in podocytes of each group; double luciferase target experiment was used to verify the targeting relationship between miR-31 and FIH-1 gene.Results Compared with the LG and HM group, the expression of miR-31 in the HG group was significantly increased(F=146.8, P<0.01), and the expression of FIH-1 was significantly decreased(F=54.23, P<0.01), and the expression of TGF-β1 and α-SMA were increased significantly(F=360.6,P<0.01; F=193.7, P<0.01). The results of the dual luciferase reporter gene experiment showed that FIH-1 was the target gene of miR-31. When si-FIH-1 and miR-31i were co-transfected, the changes in the mRNA and protein expression of FIH-1, TGF-β1, and α-SMA caused by si-FIH-1 or miR-31i alone were restored.Conclusion miR-31 can promote the EMT of podocytes by targeting FIH-1. Inhibiting miR-31 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:袁凤易,Email:1332911160@qq.com
基金项目:广东省医学科研基金项目(A2021302); 国家自然科学青年项目(81900378)
更新日期/Last Update: 2022-09-10