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本文旨在研究miR-486-5p对C2C12细胞IGF-1-PI3K/AKT-mTOR信号通路的影响,探讨miR-486-5p在C2C12细胞中对胰岛素信号通过基因的调控机制。利用构建好的miR-486-5p过表达和抑制慢病毒载体,通过脂质体转染法分别在C2C12细胞上过表达和抑制miR-486-5p的表达,利用荧光定量PCR法检测miR-486-5p过表达和抑制效率,同时检测过表达和抑制miR-486-5p后IGF-1、INSR、IRS1、PI3K、PI3KR1、PDK1、AKT1、PTEN、Fox O1、m TOR、4EBP1基因的mRNA变化。荧光定量PCR结果显示24 h过表达倍数和抑制效率分别为2.8倍和35%,48 h过表达倍数和抑制效率分别为1.4倍和15%。过表达miR-486后,24 h和48 h时INSR、PDK1、PTEN、AKT1、FoxO1、4EBP1基因均显著下调;抑制miR-486后,24 h时4EBP1显著上调,48 h时IGF-1显著上调,4EBP1显著下调。本研究表明过表达miR-486-5p后胰岛素信号通路基因的表达受到抑制,抑制miR-486-5p后胰岛素信号通路基因的表达上升,这可能是肥胖患者血清miR-486-5p升高损害胰岛素信号引起葡萄糖摄取能力降低,从而导致胰岛素抵抗发生的原因。
Abstract：To study the effect of miR-486-5 p on the IGF-1-PI3 K/AKT-mTOR signaling pathway in C2 C12 cells and to investigate the regulation mechanism of miR-486-5 p on insulin signaling genes in C2 C12 cells, the miR-486-5 p was overexpressed and inhibited on C2 C12 cells by liposome transfection using the constructed miR-486-5 p overexpression and the inhibition lentivirus vector, respectively. The overexpression and the inhibition efficiency of miR-486-5 p were detected by RT-qPCR. Changes of IGF-1, INSR, IRS1, PI3 K, PI3 KR1, PDK1,AKT1, PTEN, Fox O1, m TOR and 4 EBP1 mRNA expression were detected after overexpression and inhibition of miR-486-5 p. RT-qPCR showed that the efficiency of overexpression and inhibition were 2.8 and 35% at 24 h, 1.4 and 15% at 48 h, respectively. After overexpression of miR-486-5 p, INSR, PDK1, PTEN, AKT1, FoxO1 and 4 EBP1 genes significantly decreased at 24 h and 48 h; after inhibition of miR-486-5 p, 4 EBP1 significantly increased at 24 h, IGF-1 significantly increased but 4 EBP1 significantly decreased at 48 h. The expression of insulin signaling pathway genes was up-regulated after miR-486-5 p overexpression and down-regulated after miR-486-5 p inhibition. It may be the reason that the increase of serum miR-486-5 p in obese patients impaired the insulin signaling and caused the decrease of glucose uptake and insulin resistance.
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基本信息:
DOI：10.13417/j.gab.040.000481
中图分类号:R587.1
引用信息:
[1]郭晓萍,陈时锦,张名媛,等.miR-486-5p对C2C12细胞IGF-1-PI3K/AKT-mTOR信号通路的影响[J].基因组学与应用生物学,2021,40(02):481-488.DOI:10.13417/j.gab.040.000481.
基金信息:
广西自然科学基金项目(2018GXNSFBA281002); 广西高校中青年教师基础能力提升项目(2018KY0108)共同资助
2021-02-25
2021-02-25