基础医学与临床 ›› 2019, Vol. 39 ›› Issue (7): 973-977.

• 研究论文 • 上一篇    下一篇

miR-17-5p诱导小鼠肾足细胞系凋亡

任玮1,余宏川1,王萍2,刘英茹1   

  1. 1. 西安市儿童医院
    2. 陕西省人民医院消化内科
  • 收稿日期:2018-06-01 修回日期:2018-11-21 出版日期:2019-07-05 发布日期:2019-07-02
  • 通讯作者: 任玮 E-mail:renwei2272@163.com

miR-17-5p induces podocytes apoptosis and inhibits glomerular epithelial protein 1 in mice

  • Received:2018-06-01 Revised:2018-11-21 Online:2019-07-05 Published:2019-07-02

摘要: 目的 探讨miR-17-5p在小儿肾病综合征 (NS)发病中的作用及其机制。方法 将miR-17-5p mimic转染入小鼠肾足细胞系MPC5 24 h后收集细胞。通过microRNA.org、Target Scan和PicTar在线预测miR-17-5p对PTPRO-3' UTR区的调控作用。用RT-PCR和Western blot检测PTPRO mRNA以及蛋白的表达, Fluo-3-Am特异性Ca2+荧光指示剂检测足细胞内游离钙离子浓度([Ca2+]i), annexin V/PI双染色流式细胞计量术检测足细胞凋亡率。结果 生物信息学技术预测,小鼠源还以及人源miR-17-5p都有多个位点结合于相应的PTPRO-3' UTR区,并抑制PTPRO蛋白及mRNA的表达。PTPRO过表达抑制miR-17-5p诱导的足细胞[Ca2+]i升高,PTPRO过表达减缓miR-17-5p mimic诱导的肾小球足细胞凋亡。结论miR-17-5p抑制小鼠肾足细胞内PTPRO表达, 并激发肾小球足细胞[Ca2+]i升高,诱导肾小球足细胞凋亡。

关键词: MiR-17-5p, 受体型酪氨酸磷酸酯酶O, 肾小球足细胞, 凋亡

Abstract: Objective To explore the role and mechanisms of miR-17-5p in the pathogenesis of childhood nephrotic syndrome. Methods miR-17-5p mimic were transfected into podocytes in kidney of mice (MPC5) for 24 h to harvest the cells. the potential targets of MiR-117-5P were searched using the prediction programs microRNA.org, TargetScan and PicTar. RT-PCR and Western blot were used to detect expression of PTPRO mRNA and protein. Ca2+ fluorescence indicator with Fluo-3-Amspecificity were used to measure intracellular free calcium concentration ([Ca2+]i) in podocytes. Annexin V/PI flow cytomeutry was used to test apoptosis rate of podocytes. Results According to the bioinformatics technology prediction, miR-17-5p derived from human or mouse all can bind to multiple sites of the corresponding PTPRO-3 'UTR region and inhibit the expression of PTPRO protein and mRNA. PTPRO overexpression inhibits miR-17-5p-induced increase of [Ca2+]i in podocytes in kidney of mice. PTPRO overexpression alleviates miR-17-5p-induced apoptosis of podocytes in kidney of mice. Conclusion miR-17-5p inhibits expression of PTPRO in podocytes in kidney of mice and stimulates increase of intracellular free calcium concentration ([Ca2+]i) in podocytes, inducing apoptosis of podocytes in kidney of mice.

Key words: MiR-17-5p, glomerular epithelial protein 1, glomerular podocytes, apoptosis