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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 palmitate-induced insulin resistance (IR) in HepG2 cells and its relationship with nuclear factor-κB , NF-κB) relationship. Methods HepG2 cells were divided into normal control group, palmitic acid group (plus 0.25 mmol / L palmitate), high insulin group (plus 100 nmol / L insulin), palmitic acid + genipin group Palmitic acid and 10 μmol / L genipin). After cultured for 24 h, the concentrations of glucose, MDA, SOD, ALT, AST, GGT and TG were determined after 12 h stimulation with 100 nmol / L insulin. O staining, the mitochondrial membrane potential was detected by flow cytometry. After 30min, the expression of IRS-2, UCP-2 and NF-κB were detected by semi-quantitative RT-PCR and Western blotting. Results After 12 h of insulin treatment, there was no significant difference in glucose content between palmitate and insulin group (P> 0.05). The content of glucose, ALT, AST, MDA, GGT, TG and the expression of UCP-2 and NF-κB in the culture medium were significantly higher than those in the normal control group and palmitic acid + genipin group (P <0.05). There was no significant difference between palmitic acid + genipin group and normal control group (P> 0.05). Mitochondrial membrane potential, SOD and IRS-2 levels were significantly lower in the palmitate group than those in the normal control group and palmitate + genipin group (P <0.05), and palmitate + genipin group compared with the normal control group The difference was not statistically significant (P> 0.05). Conclusion UCP-2 plays an important role in palmitate-induced hepatic insulin resistance, and its mechanism may be related to NF-κB.