核糖体蛋白S6激酶2沉默在乙醇诱导的心肌胰岛素抵抗中的作用

doi:10.6040/j.issn.1671-7554.0.2019.1388

摘要/Abstract

摘要： 目的探讨核糖体蛋白S6激酶2(S6K2)沉默在乙醇诱导的心肌胰岛素抵抗中的作用。方法采用荧光标记2-脱氧葡萄糖(2-NDBG)摄取实验及葡萄糖氧化酶法检测在100 mmol乙醇刺激下,沉默S6K2前后心肌细胞H9c2糖摄取能力。采用Western blotting法检测乙醇处理及病毒转染前后S6K2蛋白表达量及胰岛素受体底物 1(p-IRS-1)磷酸化情况。采用q-PCR及Western blotting法检测慢病毒转染后S6K2的沉默情况。结果乙醇刺激降低了心肌细胞H9c2葡萄糖摄取,2-NDBG平均荧光强度降低46.5%(P=0.016),同时p-IRS-1(Ser)蛋白表达量提高14.4%(P=0.024),S6K2在mRNA及蛋白水平表达分别上调16.8%和23.5%(P=0.009,P=0.029)。S6K2沉默后,乙醇刺激对H9c2心肌细胞糖摄取及p-IRS-1(Ser)的表达差异无统计学意义(P=0.665,P=0.740)。结论乙醇刺激导致心肌细胞糖摄取障碍,诱导胰岛素抵抗,S6K2沉默能够有效改善乙醇诱导的心肌细胞葡萄糖摄取障碍及胰岛素抵抗现象。

关键词: 乙醇, 心肌, 胰岛素抵抗, H9c2心肌细胞, 核糖体蛋白S6激酶2

Abstract: Objective To explore the role of ribosomal protein S6 kinase 2(S6K2)in ethanol-induced myocardial insulin resistance. Methods The glucose uptake capacity of H9c2 cells after 100 mmol ethanol stimulation and lentivirus transfection was detected with fluorescence labeling 2-deoxy-2-［(7-nitro-2,1,3-benzoxadiazol-4-yl)amino］-D-glucose(2-NDBG)intake test and glucose oxidase. The expression of S6K2 and phosphorylation of insulin receptor substrate 1(p-IRS-1)were determined with Western blotting. The silencing of S6K2 after lentivirus transfection was detected with real time PCR and Western blotting. Results Ethanol exposure impaired glucose uptake in H9c2 cells. The mean fluorescence intensity of 2-NDBG decreased by 46.5%(P=0.016), the protein expression of p-IRS-1(Ser)increased by 14.4%(P=0.024), and the mRNA and protein expressions of S6K2 increased by 16.8% and 23.5% respectively(P=0.009, P=0.029). After S6K2 silencing, the difference of glucose uptake and p-IRS-1(Ser)expression in ethanol-stimulated H9c2 cells had no statistics significance(P=0.665, P=0.740). Conclusion Ethanol exposure triggers glucose intolerance and insulin resistance in H9c2 cells, and S6K2 silencing can alleviate impaired glucose uptake and insulin sensitivity induced by ethanol.

Key words: Ethanol, Myocardium, Insulin resistance, H9c2 cardiomyocyte, Ribosomal protein S6 kinase 2

中图分类号:

R542.2

徐红照,郝宗山, 沈德嫱,李朝苹,陈立勇. 核糖体蛋白S6激酶2沉默在乙醇诱导的心肌胰岛素抵抗中的作用[J]. 山东大学学报 (医学版), 2020, 58(4): 84-89.

XU Hongzhao, HAO Zongshan, SHEN Deqiang, LI Zhaoping, CHEN Liyong. Ribosomal protein S6 kinase 2 silencing in ethanol-induced myocardial insulin resistance[J]. Journal of Shandong University (Health Sciences), 2020, 58(4): 84-89.

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