miR-431靶向作用IRS2调控脂肪细胞分化及参与胰岛素抵抗发生的机制研究

项目名称： miR-431靶向作用IRS2调控脂肪细胞分化及参与胰岛素抵抗发生的机制研究

项目编号： No.81460170

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 曹玲玲

作者单位： 南昌大学

项目金额： 48万元

中文摘要： 项目以miRNA为视角，基于脂肪细胞分化是胰岛素抵抗发生发展的细胞生理学基础这一理论，朝着阐述miR-431靶向IRS2调控脂肪细胞分化及参与胰岛素抵抗发生机制的目标开展工作。在预实验筛选到脂肪细胞分化过程显著差异的miR-431及其候选靶基因胰岛素受体底物蛋白2(IRS2)的基础上，运用定量PCR、免疫印迹、荧光素酶和慢病毒重组载体感染等技术观察miR-431和IRS2在3T3-L1脂肪细胞分化中的表达变化及功能的相关性，并通过靶位点鉴定，来证实miR-431靶向IRS2调控脂肪细胞分化；再通过建立体内体外胰岛素抵抗模型，观察miR-431对胰岛素抵抗中脂肪细胞分化能力、糖代谢和胰岛素敏感性的影响，证实miR-431靶向IRS2参与胰岛素抵抗发生。本项目从多角度系统阐明miR-431靶向IRS2调控脂肪细胞分化及参与胰岛素抵抗发生的机制，为深入认识糖尿病和肥胖症等疾病提供理论依据。

中文关键词： 微小RNA431；胰岛素受体底物蛋白2；脂肪细胞分化；胰岛素抵抗

英文摘要： Based on the theory that impaired adipocyte differentiation contributes to insulin resistance, this study further explains that microRNA-431 (miR-431) regulates adipocyte differentiation and plays a role of insulin resistance by targeting Insulin Receptor Substance 2 (IRS2). In the previous experiements, markably differential miR-431 and candidate target gene IRS2 are selected in the process of adipocyte differentiation. After treated with technqiues such as Realtime PCR, immunoblotting test and Persistent Viral Recombination Carrier, the correlations between miR-431 and IRS2 are observed in gene expressions and functions during the 3T3-L1 adipocytes differentiation, and the binding target spot of miR-431 is identified to existe in the 3'UTR of IRS2 gene by the SuperLight Luciferase Reporter Gene assay, which verify that miR-431 regulates the adipocyte differentiation by suppressing IRS2, a direct target of miR-431. The influence of miR-431 on adipocyte differentiation, glucose metabolism, and insulin sensitivity is observed in vivo and vitro insulin-resistance model to support the assumption that miR-431 is involved in the development of insulin resistance by targeting IRS2. This study is going to explore the role of miR-431 in regulation of adipocyte differentiation and development of insulin resistance by targeting IRS2 in multiple aspects and provides theoretical foundation to further recognize the the diseases like diabetes and obesity.

英文关键词： miR-431;IRS2;adipogenic differentiation;insulin resistance