解偶联蛋白-2在软脂酸诱导的HepG2细胞胰岛素抵抗中的作用及其与核转录因子-κB的关系  

Role of Uncoupling Protein-2 in Insulin-resistance Induced by Palmic Acid in HepG2 Cells and Its Relationship to Nuclear Factor-κB

作　　者：王立娟[1] 管小琴[1] 刘琳[1] 唐袁婷[1] 朱良荣[1] 

机构地区：[1]重庆医科大学病理教研室分子医学与肿瘤研究中心,重庆400016

出　　处：《中国生物制品学杂志》2010年第8期807-812,共6页Chinese Journal of Biologicals

基　　金：重庆市教委资助项目(KJ090327)

摘　　要：目的探讨解偶联蛋白-2(Uncoupling protein-2,UCP-2)在软脂酸诱导的HepG2细胞胰岛素抵抗(Insulin-resistance,IR)中的作用及其与核转录因子-κB(Nuclearfactor-κB,NF-κB)的关系。方法将HepG2细胞分为正常对照组、软脂酸组(加0.25mmol/L软脂酸)、高胰岛素组(加100nmol/L胰岛素)、软脂酸+京尼平组(加0.25mmol/L软脂酸和10μmol/L京尼平),培养24h后,再用100nmol/L胰岛素刺激,分别于12h后测定培养液中葡萄糖、MDA、SOD、ALT、AST、GGT、TG的浓度;油红O染色法观察细胞的脂变情况;流式细胞术检测线粒体膜电位改变;30min后采用半定量RT-PCR及Westernblot法检测细胞IRS-2、UCP-2及NF-κB的表达水平。结果胰岛素作用12h后,细胞培养液中葡萄糖含量软脂酸组与高胰岛素组比较,差异无统计学意义(P>0.05)。培养液中葡萄糖含量、ALT、AST、MDA、GGT、TG的含量及UCP-2和NF-κB的表达水平,软脂酸组均较正常对照组和软脂酸+京尼平组显著升高(P<0.05),软脂酸+京尼平组与正常对照组比较差异无统计学意义(P>0.05)。线粒体膜电位、SOD和IRS-2的水平,软脂酸组显著低于正常对照组和软脂酸+京尼平组(P<0.05),软脂酸+京尼平组与正常对照组比较差异无统计学意义(P>0.05)。结论 UCP-2在软脂酸诱导的肝脏胰岛素抵抗中起着重要作用,其机制可能与NF-κB有关。Objective To investigate the role of uncoupling protein-2（UCP-2）in inulin-resistance induced by palmic acid（PA） in HepG2 cells and its relationship to nuclear factor-κB（NF-κB）. Methods HepG2 cells were divided into normal control, PA, high insulin（HI）and PA ＋ Genipin groups. The cells in normal control groups were cultured in RPMI1640 medium containing 10 calf serum, while those in PA, HI and PA ＋ Genipin groups in the media added with 0. 25 mmol / L PA, 100 nmol / L insulin and 0. 25 mmol / L PA ＋ 10 μmol / L Genipin respectively, for 24 h, then stimulated with 100 nmol / L insulin. The glucose, MDA, SOD, ALT, AST, GGT and TG concentrations in media were determined, the fatty degeneration of cells was observed by oil red O staining, the change of mitochondrial membrane potential was analyzed by flow cytometry, and the expression levels of IRS-2, UCP-2 and NF-κB were determined by RT-PCR and Western blot. Results The glucose contents in media of PA and HI groups 12 h after stimulation with insulin showed no significant difference （P 0. 05）. The glucose, ALT, AST, MDA, GGT and TG contents as well as the expression levels of UCP-2 and NF-κB were significantly higher in PA group than in normal control and PA ＋ Genipin groups （P 0. 05）, while showed no significant difference in PA ＋ Genipin and normal control groups（P 0. 05）. The mitochondrial membrane potential as well as SOD and IRS-2 levels were significantly lower in PA group than in normal control and PA ＋ Genipin groups （P 0. 05）, while showed no significant difference in PA ＋ Genipin and normal control groups （P 0. 05）. Conclusion UCP-2 played an important role in the inulin-resistance induced by PA in HepG2 cells, of which the mechanism might be related to NF-κB.

关 键 词：解偶联蛋白-2 胰岛素抵抗 京尼平 胰岛素受体底物-2 核转录因子-ΚB 

分 类 号：R977.4[医药卫生—药品]