基础医学与临床 ›› 2023, Vol. 43 ›› Issue (2): 271-276.doi: 10.16352/j.issn.1001-6325.2023.02.271

• 研究论文 • 上一篇    下一篇

过表达miR-210抑制H2O2诱导大鼠BM-MSCs成脂分化

封宝红, 喻业安*, 晏莉, 伍军, 张艳霞, 朱戈丽   

  1. 武汉大学附属武汉市第三医院 肾内科, 湖北 武汉 430014
  • 收稿日期:2022-03-16 修回日期:2022-07-03 出版日期:2023-02-05 发布日期:2023-02-02
  • 通讯作者: *yuyean@163.com
  • 基金资助:
    武汉市卫生健康委员会资助项目(WX20B22)

Over-expression of miR-210 inhibits H2O2-induced adipogenic differentiation of rat BM-MSCs

FENG Baohong, YU Yean*, YAN Li, WU Jun, ZHANG Yanxia, ZHU Geli   

  1. Department of Nephrology, the Third Affiliated Hospital of Wuhan University, Wuhan 430014, China
  • Received:2022-03-16 Revised:2022-07-03 Online:2023-02-05 Published:2023-02-02
  • Contact: *yuyean@163.com

摘要: 目的 探讨miR-210对过氧化氢(H2O2)诱导大鼠骨髓间充质干细胞(BM-MSCs)成脂分化的影响。方法 分离培养并鉴定SD大鼠BM-MSCs细胞。200 μmol/L H2O2诱导BM-MSCs 1 h以构建成脂分化模型细胞,分别利用miR-210 模拟物或抑制物转染成脂分化模型细胞,RT-qPCR检测miR-210表达;油红O染色检测细胞胞质内的脂肪颗粒;Western blot检测细胞中过氧化物酶体增殖物激活受体γ(PPARγ)、CCAAT/强子结合蛋白α(C/EBPα)及Wnt/β-连环蛋白(β-catenin)通路蛋白。结果 成功分离SD大鼠BM-MSCs细胞。成脂分化模型BM-MSCs中miR-210、β-catenin、c-Myc、细胞周期蛋白 D1(cyclin D1)蛋白水平降低,脂肪颗粒数目、PPARγ、C/EBPα蛋白水平升高;miR-210模拟物转染的BM-MSCs中miR-210、β-catenin、c-Myc、cyclin D1蛋白水平升高,脂肪颗粒数目、PPARγ、C/EBPα 蛋白水平降低;miR-210 抑制物转染的BM-MSCs中对应指标变化趋势与miR-210模拟物转染的BM-MSCs相反。结论 过表达miR-210可抑制H2O2诱导的BM-MSCs成脂分化,机制可能与激活Wnt/β-catenin通路有关。

关键词: miR-210, 过氧化氢, 骨髓间充质干细胞, 成脂分化

Abstract: Objective To investigate the effect of miR-210 on hydrogen peroxide (H2O2)-induced adipogenic differentiation of rat bone marrow mesenchymal stem cells (BM-MSCs). Methods The SD rat BM-MSCs were isolated, cultured and identified. BM-MSCs were induced by 200 μmol/L H2O2 for 1 h to construct an adipogenic differentiation model. Adipogenic differentiation models were transfected with miR-210 mimic or inhibitor, respectively. RT-qPCR to detect the expression of miR-210; oil red O staining to detect the fat particles in the cytoplasm of cells; Western blot to detect the expression of peroxisome proliferator-activated receptor γ (PPARγ), CCAAT enhancer binding protein (C/EBPa) and Wnt/β-catenin pathway related proteins of cells. Results SD rat BM-MSCs were successfully isolated. In adipogenic differentiation model of BM-MSCs, the levels of miR-210, β-catenin, c-Myc, and cyclin D1 protein decreased, while the number of fat granules, PPARγ and C/EBPα protein levels increased; In BM-MSCs transfected with miR-210 mimic, the levels of miR-210, β-catenin, c-Myc, and cyclin D1 protein increased, while the fat granules, PPARγ, and C/EBPα protein level all decreased; The change trend of corresponding indexes in BM-MSCs transfected with miR-210 inhibitor was opposite to that in BM-MSCs transfected with miR-210 mimic. Conclusions Over-expression of miR-210 could inhibit the adipogenic differentiation of BM-MSCs induced by H2O2. This mechanism may be related to the activation of Wnt/β-catenin pathway.

Key words: miR-210, hydrogen peroxide, bone marrow mesenchymal stem cells, adipogenic differentiation

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