【摘要】 目的:观察肿瘤坏死因子α和噻唑烷二酮类药物吡格列酮对3T3-L1脂肪细胞中葡萄糖转运子4(glucose transporter4,GLUT4)mRNA和蛋白表达的影响,并了解其是否有时间依赖性。方法:实验于2005-09/2006-05在安徽医科大学病原微生物分子生物学教研室进行。对3T3-L1细胞进行培养并诱导分化为成熟的3T3-L1脂肪细胞后随机分为对照组、肿瘤坏死因子α组和吡格列酮组3组。肿瘤坏死因子α组和吡格列酮组分别加含20μg/L肿瘤坏死因子α或10-4mol/L吡格列酮培养基培养,提取干预后1,3,6d细胞总RNA和总蛋白做RT-PCR和Western-blot,测定GLUT4mRNA和蛋白的表达,以未加任何药物干预组做对照。结果:①GLUT4mRNA表达:对照组细胞为0.506±0.049;肿瘤坏死因子α组干预后1,3,6d表达低于对照组(0.465±0.039,0.410±0.010,0.320±0.019,F=17.8,P=0.001),并随干预时间呈递减关系;吡格列酮组干预后1,3,6d表达高于对照组(0.544±0.064,0.616±0.065,0.664±0.070,F=4.87,P=0.043),且与干预时间呈正比。②GLUT4蛋白的相对含量:对照组细胞为0.624±0.093;肿瘤坏死因子α组干预后1,3,6d低于对照组(0.549±0.112,0.460±0.111,0.286±0.117,F=7.39,P=0.011),且随干预时间递减;而吡格列酮组GLUT4干预后1,3,6d高于对照组(0.693±0.098,0.750±0.106,0.866±0.074,F=4.0,P=0.048),随干预时间呈递增关系。结论:①肿瘤坏死因子α可降低3T3-L1脂肪细胞中GLUT4mRNA和蛋白表达量,吡格列酮的作用与肿瘤坏死因子α相反,两者作用均呈时间依赖性。②在3T3-L1脂肪细胞中,吡格列酮可能通过增加GLUT4的表达来提高胰岛素敏感性,并可能部分拮抗肿瘤坏死因子α诱导的胰岛素抵抗。更多还原

【Abstract】 AIM:To investigate the effects of tumor necrosis factor-α(TNFα)and pioglitazone on glucose transporter 4(GLUT4)mRNA and protein expression in 3T3-L1 adipocytes,and explore the time dependence.METHODS:The experiment was completed in the Research Room of Pathogenic Microorganism and Molecular Biology of Anhui Medical University from September 2005 to May 2006.The differentiated 3T3-L1 adipocyte cells were divided randomly into three groups:control group,TNFα group and pioglitazone group.20 μg/L TNFα and 10-4 mol/L pioglitazone were adopted in the TNFα group and pioglitazone group,respectively.After intervening for 1,3 and 6 days,total RNA and total protein received RT-PCR and Western-blot to determine the expressions of GLUT4mRNA and protein,and then compared with the control group.RESULTS:①GLUT4mRNA expression:It was 0.506±0.049 in the control group.It was lower in the TNFα group after intervening for 1,3 and 6 days(0.465±0.039,0.410±0.010,0.320±0.019,F =17.8,P =0.001),and decreased gradually with time dependence.It was higher in the pioglitazone group after intervening for 1,3 and 6 days than the control group(0.544±0.064,0.616±0.065,0.664±0.070,F =4.87,P =0.043),and showed direct proportion with the time of intervention.②Relative amount of GLUT4:It was 0.624±0.093 in the control group.It was lower in the TNFα group after intervening for 1,3 and 6 days(0.549±0.112,0.460±0.111,0.286±0.117,F =7.39,P =0.011),and decreased gradually with time dependence.It was higher in the pioglitazone group after intervening for 1,3 and 6 days than the control group(0.693±0.098,0.750±0.106,0.866±0.074,F =4.0,P =0.048),and showed increase with the time of intervention.CONCLUSION:①TNFα can decrease the contents of GLUT4mRNA and protein expression in 3T3-L1 adipocytes.TNFα and pioglitazone have opposite effects that have time dependence.②Pioglitazone appeares to improve insulin sensitivity by increasing the GLUT4 expression and partly inhibits the effect of TNFα on insulin resistance in 3T3-L1 adipocytes.更多还原