抵抗素对大鼠肝细胞磷酸烯醇式丙酮酸羧激酶酶活性及mRNA表达的影响  

Effect of resistin on the activity and gene expression of phosphoenolpyruvate carboxykinase in cultured rat liver cells

作　　者：李超林[1] 赵璐杰[2] 肖谦[3] 

机构地区：[1]山东省烟台市毓璜顶医院内分泌科,烟台264000 [2]烟台市毓璜顶医院血液透析中心 [3]重庆医科大学附属第一医院老年科

出　　处：《临床医学》2012年第9期103-105,共3页Clinical Medicine

摘　　要：目的探讨抵抗素对大鼠肝细胞糖异生代谢关键酶-磷酸烯醇式丙酮酸羧激酶(PEPCK)活性及mRNA表达的影响,了解抵抗素对肝脏糖代谢的影响并初步探讨抵抗素在肝脏胰岛素抵抗形成中的作用及机制。方法正常大鼠的肝细胞,不加抵抗素与加不同剂量的抵抗素(10、50、100 nmol/L)培养24 h。乳酸脱氢酶偶联比色法检测不同剂量抵抗素对PEPCK活性的影响;逆转录-聚合酶链反应(RT-PCR)检测PEPCK mRNA表达水平的变化。结果抵抗素浓度为10、50 nmol/L时,PEPCK活性[单位为mIU/(min.mg protein)]较空白对照组显著升高(31.8±2.4,32.0±2.3 vs 26.6±2.6,P<0.05),抵抗素浓度为100 nmol/L,PEPCK活性升高更明显(40.4±1.5 vs 26.6±2.6,P<0.01)。RT-PCR结果显示抵抗素浓度为10、50 nmol/L时,PEPCK mRNA表达水平(以β-actin校正)与空白对照组比较差异无统计学意义(P﹥0.05),抵抗素浓度为100 nmol/L,PEPCK mRNA表达水平显著升高(1.18±0.12 vs 0.74±0.08,P﹤0.01)。结论抵抗素浓度为10、50 nmol/L时,PEPCK活性显著升高,但对其mRNA表达水平无显著影响。抵抗素浓度为100 nmol/L时,PEPCK活性升高更明显,同时其mRNA表达水平也显著升高。表明该浓度的抵抗素至少部分通过促进PEPCK基因的表达实现对酶活性的调节。Objective To investigate the effects of resistin on activity and gene expression of phosphoenolpyruvate carboxyki- nase （PEPCK） in cultured rat liver cells, and the role of resistin on heptic glucose metabolism and the mechanism that how resis- tin affected liver insulin resistance （ 1R ）. Methods Rat liver cells were incubated with resistin of different doses （ 10, 50, 100 nmol/L）. Controlled liver cells were incubated without resistin. Lactate dehydrogenase couple chromatometry were used to determine activity of PEPCK; RT-PCR was used to determine PEPCK gene expression （corrected by β-actin ）. Results Comparing with the control group, activity of PEPCK [ Unit is mIU/（ min·mg protein ）] increased significantly （31.8±2.4, 32. 0 ±2. 3 vs 26. 6 ±2. 6, P 〈 0. 05） afer treatment with resistin of 10 nmol/L and 50 nmoL/L for 24 h, at the dose of 100 nmol/L, activity of PEPCK increased more significantly（40. 4 ± 1.5 vs 26.6 ±2. 6, P 〈 0. 01 ）. RT-PCR result showed： Comparing with the control group, treatment with resistin of 10 nmoL/L and 50 nmol/L for 24 h had no significant effect on gene expression of PEPCK （P 〉 0. 05 ） , at the dose of 100 nmoL/L, the PEPCK mRNA expression was promoted significantly （ 1.18 -± 0.12 vs 0.74 -± 0.08, P 〈 0. 01 ）. Conclusion 10 nmol/L and 50 nmol/L of resistin increase activity of PEPCK sig- nificantly, but have little effect on gene expression of PEPCK..At the dose of 100 nmol/L, the activity of PEPCK increase more significantly and PEPCK mRNA expression is promoted significantly. This effect is at least partly brought by the promotion of PEPCK gene expression.

关 键 词：抵抗素 磷酸烯醇式丙酮酸羧激酶 肝细胞 

分 类 号：R587.1[医药卫生—内分泌]