锌α2糖蛋白通过调节脂肪细胞自噬增强胰岛素信号通路表达的作用及机制研究  

Effect of zinc α2 glycoprotein in enhancing expression of insulin signaling pathway by regulating autophagy of adipocytes and its mechnism

作　　者：张韶君[1] 遆红燕 郭亚莉 高江琴[3] 燕桂新 康芳芳 岳志远 邱贵娟 Zhang Shaojun;Ti Hongyan;Guo Yali;Gao Jiangqin;Yan Guixin;Kang Fangfang;Yue Zhiyuan;Qiu Guijuan(Department of Endocrinology,Shanxi Provincial People′s Hospital,Taiyuan 030012,China)

机构地区：[1]山西省人民医院内分泌科,太原030012 [2]山西医科大学研究生学院 [3]新疆生产建设兵团第六师医院内分泌科

出　　处：《中国药物与临床》2018年第12期2120-2122,共3页Chinese Remedies & Clinics

基　　金：山西省基础研究项目(2013011048-1);新疆生产建设兵团科技援疆计划资金项目(2014AB050)

摘　　要：目的研究锌α2糖蛋白(ZAG)对胰岛素抵抗3T3-L1细胞磷酸化的丝氨酸/苏氨酸激酶[(pAkt(Ser473)]水平和葡萄糖转运蛋白4(GLUT4)m RNA和蛋白表达的影响,探讨ZAG改善胰岛素抵抗的作用及分子机制。方法将3T3-L1脂肪细胞随机分为对照、胰岛素抵抗、ZAG、氯喹、ZAG+氯喹5组,应用地塞米松诱导细胞胰岛素抵抗,ZAG组加入ZAG,氯喹组加入氯喹,ZAG+氯喹组同时加入ZAG和氯喹。以葡萄糖氧化酶法测定5组细胞上清液中葡萄糖消耗量,观察ZAG对脂肪细胞葡萄糖摄取的影响;应用蛋白质印迹法测定脂肪细胞p-Akt (Ser473)和GLUT4的蛋白水平变化;应用实时荧光定量聚合酶链反应(PCR)技术检测脂肪细胞GLUT4的mRNA表达变化。结果与正常对照组相比,胰岛素抵抗组的p-Akt(Ser473)和GLUT4表达明显下降(P均<0.05),经ZAG干预后,p-Akt(Ser473)和GLUT4表达水平有一定升高,但仍显著低于正常对照组(P均<0.05),加入氯喹后,p-Akt(Ser473)和GLUT4表达降低(P均<0.05)。结论 ZAG通过加强脂肪细胞自噬活性,增强AKT磷酸化进而增加GLUT4的水平,改善胰岛素抵抗。Objective To determine the effect of zincα2 glycoprotein(ZAG)on the phosphorylated serine/threonine kinase[p-Akt(Ser473)]level and the mRNA and protein expression of glucose transporter 4(GLUT4)in insulin-resistant 3T3-L1 cells,and to investigate the effect of ZAG in improving insulin resistance and its molecular mechanism.Methods The 3T3-L1 adipocytes were randomly divided into the normal control group,insulin resistance group,ZAG group,chloroquine group,and ZAG+chloroquine group.Dexamethasone was used to induce insulin resistance in the cells.ZAG was added to the ZAG group,chloroquine was added to the chloroquine group,and ZAG and chloroquine were simultaneously added to the ZAG+chloroquine group.The glucose consumption in the supernatants of the five groups was determined by the glucose oxidase method,and the effect of ZAG on the glucose uptake by adipocytes was determined.The protein levels of p-Akt(Ser473)and GLUT4 protein were determined by Western blotting.Real-time fluorescence quantitative PCR was used to examine the change of mRNA expression of GLUT4 in adipocytes.Results Compared with the normal control group,the expression levels of p-Akt(Ser473)and GLUT4 significantly reduced in the insulin resistance group(P<0.05).After ZAG intervention,the expression levels of p-Akt(Ser473)and GLUT4 increased,but were still significantly lower than those in the normal control group(all P<0.05).After the addition of chloroquine,the expression levels of p-Akt(Ser473)and GLUT4 decreased(P<0.05).Conclusion ZAG enhances AKT phosphorylation by enhancing the autophagy activity of adipocytes,and thereby increases GLUT4 level and improves insulin resistance.

关 键 词：锌α2糖蛋白 自噬 蛋白质丝氨酸苏氨酸激酶 葡萄糖转运体4型 

分 类 号：R587.1[医药卫生—内分泌]