Department of Clinical Nutrition, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwu Road, Jinan, 250021, Shandong, China.
Department of Clinical Nutrition, Lianyungang Hospital Affiliated to Xuzhou Medical University, Lianyungang, 222061, Jiangsu, China.
Mol Biol Rep. 2020 Dec;47(12):9469-9477. doi: 10.1007/s11033-020-05967-6. Epub 2020 Nov 7.
Alcohol exposure impairs myocardium insulin sensitivity, which links to heart dysfunction. miR-155 regulates mTOR signaling pathway and is involved in multiple functions. However, the underlying mechanism of miR-155 in ethanol-induced myocardial insulin resistance remains unclear. Here, in this study we aimed to identify the role of miR-155 in myocardial insulin sensitivity and the involvement of mTOR pathway. H9C2 cells were cultured with or without 100 mM ethanol for 24 h. miR-155-5p inhibitor, miR-155-5p mimics or their respective negative control (inhibitor NC and mimic NC) were transfected to regulate miR-155-5p expression. mTOR signaling, including Ras homolog enriched in brain (Rheb), rapamycin insensitive companion of mTOR (Rictor) and ribosomal protein S6 kinase B2 (S6K2), was investigated by western blotting and qPCR, and insulin responsiveness was evaluated by glucose uptake and phosphorylation of insulin receptor substrate-1 (p-IRS1). The miR-155-5p level increased under ethanol exposure, accompanied by a decrease in glucose uptake, an increase in p-IRS1(ser 307) and activation of the mTOR signaling pathway in H9C2 cells. In addition, miR-155-5p downregulation decreased the glucose uptake, increased the p-IRS1(ser 307) level and activated the mTOR signaling pathway. miR-155-5p upregulation increased the glucose uptake, decreased the p-IRS1(ser 307) level and suppressed the mTOR signaling pathway. Collectively, these findings suggest miR-155-5p upregulation ameliorates myocardial insulin resistance via mTOR signaling in vitro, and miR-155-5p downregulation attenuates myocardial insulin resistance, which might become a potential therapeutic target for alcohol-induced cardiomyopathy.
酒精暴露会损害心肌胰岛素敏感性,从而导致心脏功能障碍。miR-155 调节 mTOR 信号通路,参与多种功能。然而,miR-155 在乙醇诱导的心肌胰岛素抵抗中的潜在机制尚不清楚。在本研究中,我们旨在确定 miR-155 在心肌胰岛素敏感性中的作用以及 mTOR 通路的参与。将 H9C2 细胞在有无 100mM 乙醇的条件下培养 24 小时。转染 miR-155-5p 抑制剂、miR-155-5p 模拟物或它们各自的阴性对照(抑制剂 NC 和模拟物 NC)以调节 miR-155-5p 的表达。通过 Western blot 和 qPCR 研究 mTOR 信号通路,包括 Ras 同源物富集在脑中(Rheb)、雷帕霉素不敏感的 mTOR 伴侣(Rictor)和核糖体蛋白 S6 激酶 B2(S6K2),并通过葡萄糖摄取和胰岛素受体底物-1(p-IRS1)的磷酸化来评估胰岛素反应性。在 H9C2 细胞中,乙醇暴露会增加 miR-155-5p 的水平,同时降低葡萄糖摄取,增加 p-IRS1(ser 307)的水平并激活 mTOR 信号通路。此外,下调 miR-155-5p 会降低葡萄糖摄取,增加 p-IRS1(ser 307)的水平并激活 mTOR 信号通路。上调 miR-155-5p 会增加葡萄糖摄取,降低 p-IRS1(ser 307)的水平并抑制 mTOR 信号通路。总之,这些发现表明,miR-155-5p 上调通过体外 mTOR 信号改善心肌胰岛素抵抗,下调 miR-155-5p 减轻心肌胰岛素抵抗,这可能成为酒精性心肌病的潜在治疗靶点。