miR-431 通过靶向胰岛素受体底物 2 抑制人骨髓间充质干细胞的成脂分化。

miR-431 inhibits adipogenic differentiation of human bone marrow-derived mesenchymal stem cells via targeting insulin receptor substance 2.

机构信息

Department of Endocrinology, The First Hospital of Jiujiang City, Jiujiang, 332000, China.

Jiujiang Affiliated Hospital of Nanchang University, Jiujiang, 332000, China.

出版信息

Stem Cell Res Ther. 2018 Aug 30;9(1):231. doi: 10.1186/s13287-018-0980-4.

BACKGROUND

An understanding of the mechanism underlying adipogenic differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs) will provide new therapeutic approaches for many diseases, including osteoporosis. This study aimed to investigate the role of miR-431 in adipogenic differentiation of hMSCs.

METHODS

hMSCs were induced for adipogenic differentiation and miR-431 was detected by polymerase chain reaction (PCR). hMSCs were transfected by miR-431 or small interfering RNA (siRNA) for insulin receptor substance 2 (IRS2). The expression of IRS2 was detected by PCR and Western blot analysis. The targeting of the 3'-untranslated region (UTR) of IRS2 by miR-431 was examined by luciferase assay.

RESULTS

miR-431 expression was decreased during adipogenesis of hMSCs. Overexpression of miR-431 inhibited adipogenic differentiation, accompanied by the downregulation of CCAAT/enhancer binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ), two key regulators of adipogenesis. Moreover, miR-431 decreased both protein and mRNA levels of IRS2. The expression of IRS2 was increased during adipogenic differentiation of hMSCs in conjunction with decreased levels of miR-431, and knockdown of IRS2 in hMSCs inhibited adipogenic differentiation. Luciferase assay confirmed that miR-431 targeted the 3'-UTR of IRS2 in hMSCs.

CONCLUSIONS

This is the first study to show that miR-431 inhibits adipogenic differentiation of hMSCs via targeting IRS2.

背景

了解人骨髓间充质干细胞（hMSCs）成脂分化的机制将为包括骨质疏松症在内的许多疾病提供新的治疗方法。本研究旨在探讨 miR-431 在 hMSCs 成脂分化中的作用。

方法

诱导 hMSCs 向成脂分化，并通过聚合酶链反应（PCR）检测 miR-431 的表达。通过 miR-431 或小干扰 RNA（siRNA）转染 hMSCs 胰岛素受体底物 2（IRS2）。通过 PCR 和 Western blot 分析检测 IRS2 的表达。通过荧光素酶测定法检测 miR-431 对 IRS2 的 3'-非翻译区（UTR）的靶向作用。

结果

miR-431 的表达在 hMSCs 成脂分化过程中降低。miR-431 的过表达抑制成脂分化，伴随着关键成脂调节因子 CCAAT/增强子结合蛋白α（C/EBPα）和过氧化物酶体增殖物激活受体γ（PPARγ）的下调。此外，miR-431 降低了 IRS2 的蛋白和 mRNA 水平。IRS2 在 hMSCs 成脂分化过程中的表达增加，同时 miR-431 的水平降低，hMSCs 中 IRS2 的敲低抑制了成脂分化。荧光素酶测定证实 miR-431 靶向 hMSCs 中的 IRS2 3'-UTR。