吡格列酮改善胰岛素抵抗状态下氧化应激水平及分子机制探讨  

Approach of the molecular mechanisms and the oxidative stress levels on pioglitazone improving insulin resistance condition

作　　者：李兰芳[1,2] 黄秀清[2] 原慧萍[2] 李淼[2] 王抒[2] 满永[2] 黎健[2] 

机构地区：[1]北京协和医学院研究生院,北京100730 [2]卫生部北京医院老年医学研究所/卫生部老年医学重点实验室,北京100730

出　　处：《中国临床药理学与治疗学》2009年第3期245-249,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：国家重点基础研究发展规划项目(973计划)(2006CB503910);国家自然科学基金(30572082)

摘　　要：目的:探讨吡格列酮(Pio)对胰岛素抵抗状态下的氧化应激水平的影响,并初步探讨Pio改善胰岛素抵抗作用的分子机制。方法:用TNF-α(4ng/mL)刺激人肝癌细胞HepG2,建立胰岛素抵抗细胞模型。MTT法观察细胞毒性作用;氧化酶法检测细胞培养液中的葡萄糖浓度;用DCFH-DA荧光探针标记,流式细胞仪检测细胞内活性氧(ROS)的水平;Western blotting检测氨基末端激酶(JNK)、磷酸化氨基末端激酶(p-JNK)和胰岛素受体底物1(IRS1)的表达。结果:TNF-α刺激HepG2后,导致葡萄糖摄取障碍,细胞培养液上清中葡萄糖浓度显著升高,细胞内ROS的水平显著升高,JNK被激活,IRS1的表达降低。Pio可显著降低ROS的水平,抑制JNK的磷酸化,促进IRS1的表达,改善胰岛素抵抗。结论:Pio通过降低氧化应激水平,抑制JNK信号通路,改善胰岛素抵抗状态。AIM： To investigate the effects of pio- glitazone on oxidative stress levels and insulin resis- tance and initially approach the molecular mechanisms of pioglitazone improving insulin resistance. METHODS： The insulin resistance cell models were induced by TNF-α （4 ng/mL）to stimulate human hepatoma car- cinoma cell HepG2. The cytotoxicity of HepG2 was ob- served by MTF. The concentrations of glucose in cell culture fluid of HepG2 were detected using a glucose oxidase assay kit. The levels of intracellular reactive oxygen species were detected by DCFH-DA fluorescent probe and flow cytometry. The expressions of JNK, p- JNK, IRS1 were measured by Western blotting. RE- SULTS： After HepG2 was stimulated by TNF-α, the glucose uptake was blocked, the glucose concentrations were significantly increased in cell culture fluid, the active oxygen levels were significantly increased in cells, the JNK was activated, the expression of IRS1 was de- creased. Pioglitazone could significantly decrease the active oxygen levels, inhibit JNK phosphorylation, pro- mote the expression of IRS1, improve insulin resistance. CONCLUSION： Pioglitazone could improve the insulin resistance condition by degrading oxidative stress levels and inhibiting JNK passway.

关 键 词：胰岛素抵抗 氧化应激 TNF-Α 

分 类 号：R587[医药卫生—内分泌]