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  

Ribosomal protein S6 kinase 2 silencing in ethanol-induced myocardial insulin resistance

作　　者：徐红照 郝宗山[2] 沈德嫱 李朝苹 陈立勇[4] XU Hongzhao;HAO Zongshan;SHEN Deqiang;LI Zhaoping;CHEN Liyong(Department of Toxicology and Nutrition,School of Public Health,Shandong University,Jinan 250012,Shandong,China;Shandong Provincial ENT Hospital Affiliated to Shandong University,Jinan 250021,Shandong,China;Department of Clinical Nutrition,Lianyungang Hospital Affiliated to Xuzhou Medical University,Lianyungang 222002,Jiangsu,China;Department of Nutrition,Shandong Provincial Hospital Affiliated to Shandong University,Jinan 250021,Shandong,China)

机构地区：[1]山东大学公共卫生学院卫生毒理与营养学系,山东济南250012 [2]山东大学附属山东省耳鼻喉医院,山东济南250021 [3]徐州医科大学附属连云港医院临床营养科,江苏连云港222002 [4]山东大学附属省立医院营养科,山东济南250021

出　　处：《山东大学学报（医学版）》2020年第4期84-89,共6页Journal of Shandong University:Health Sciences

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。Objective To explore the role of ribosomal protein S6 kinase 2(S6 K2)in ethanol-induced myocardial insulin resistance.Methods The glucose uptake capacity of H9 c2 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 S6 K2 and phosphorylation of insulin receptor substrate 1(p-IRS-1)were determined with Western blotting.The silencing of S6 K2 after lentivirus transfection was detected with real time PCR and Western blotting.Results Ethanol exposure impaired glucose uptake in H9 c2 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 S6 K2 increased by 16.8%and 23.5%respectively(P=0.009,P=0.029).After S6 K2 silencing,the difference of glucose uptake and p-IRS-1(Ser)expression in ethanol-stimulated H9 c2 cells had no statistics significance(P=0.665,P=0.740).Conclusion Ethanol exposure triggers glucose intolerance and insulin resistance in H9 c2 cells,and S6 K2 silencing can alleviate impaired glucose uptake and insulin sensitivity induced by ethanol.

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 

分 类 号：R542.2[医药卫生—心血管疾病]