耐力训练诱导AMPK对小鼠骨骼肌PGC-1/ERRα结合的影响
首发时间:2012-12-31
摘要:目的:作为能量代谢的关键调节因子,转录辅激活因子PGC-1能与ERRα结合,共同调节靶基因的转录。本研究旨在探讨耐力训练中骨骼肌细胞内AMPKα2对核内PGC-1、ERRα蛋白表达水平和PGC-1/ERRα结合量的影响。 方法:C57BL/6J野生型(WT)小鼠(n=20)、AMPKα2转基因(TG)小鼠(n=20)和AMPKα2基因敲除(KO)小鼠(n=20)三种基因状态小鼠,分别分为安静对照组(Control,C)和耐力训练组(Endurance Training,T)。耐力训练组小鼠进行持续4周,每周周一至周六,连续6天,每天1小时的跑台训练,跑台坡度为0,速度是12米/分。安静对照组小鼠在相同环境下饲养,但不参与训练。最后一次训练后16小时取小鼠骨骼肌,采用蛋白印迹法测定核内ERRα和PGC-1的蛋白表达量,免疫共沉淀法测定核内PGC-1/ERRα结合量。 结果:1)4周耐力训练后,三种基因状态小鼠骨骼肌细胞核内PGC-1、ERRα蛋白表达量和PGC-1/ERRα结合量均高于相应的安静组。2) AMPKα2转基因鼠在运动前后,PGC-1以及PGC-1/ERRα结合量都高于野生鼠。3) AMPKα2基因敲除鼠与野生鼠比较,PGC-1、 ERRα及PGC-1/ERRα结合量都没有差异。 结论:1)四周耐力训练能提高AMPKα2三种基因状态小鼠骨骼肌细胞核内PGC-1、ERRα蛋白表达量以及PGC-1/ERRα结合量。2)AMPKα2转基因能引起安静状态和耐力训练状态下,小鼠骨骼肌细胞核内PGC-1蛋白表达量和PGC-1/ERRα结合量显著性增加。3)AMPKα2基因敲除对小鼠骨骼肌细胞核内PGC-1、ERRα蛋白表达以及PGC-1/ERRα结合量影响不大。总之,AMPK是耐力运动中调节PGC-1和ERRα蛋白表达以及PGC-1/ERRα结合量的通路之一。
关键词: AMPKα2 PGC-1 ERRα 耐力运动 COIP
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Effects of Endurance Training Induced AMPKα2 on the combination of PGC-1/ERRα complex in Skeletal Muscle of Mice
Abstract:Objective The transcriptional coactivator PGC-1, a key regulator of energy metabolism, combines ERRα to induce target gene expression jointly. Here we aimed to investigate the effects of AMPKα2 on nuclear protein levels of PGC-1 and combination of PGC-1 and ERRα in skeletal muscle during endurance training. Methods C57BL/6J Wild-type(WT)mice, AMPKα2 transgenic (TG) mice and AMPKα2 knockout (KO) mice were divided into control group and training group respectively. Training mice performed 60min/day treadmill running at the speed of 12m/min for 4 weeks. Control mice were raised in the same environment without training. Skeletal muscle was collected 16 hours after the last training. Nuclear protein levels of PGC-1 were detected by Western blotting. Interaction of PGC-1 and ERRα was analyzed by co-immunoprecipitation (CoIP). Results 1) After 4 weeks endurance training, nuclear protein levels of PGC-1、ERRα and PGC-1/ ERRα complex of the three training groups were higher than their control groups respectively. 2) Control and endurance training group of TG mice exhibited higher levels of PGC-1 and PGC-1/ERRα complex compared to WT mice. 3)Both control and endurance training groups of KO mice had no difference of PGC-1、ERRα and PGC-1/ERRα complex comparing to WT mice. Conclusion 1)Endurance training increases the levels of nuclear PGC-1 and ERRα and PGC-1/ERRα complex of WT/TG/KO mice. 2) Transgene of AMPKα2 heightens the expression of nuclear PGC-1 and the level of PGC-1/ERRα complex during endurance training. 3) Knockout of AMPKα2 has no influence on the expression of PGC-1 and ERRα and PGC-1/ERRα complex .In a word, AMPK is one of the ways to regulate PGC-1 and ERRα and their combination during endurance training.
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