基于AMPK信号通路的经皮穴位电刺激对运动性骨骼肌疲劳的作用及机制研究

In recent years, the domestic and foreign research found that AMPK play a key role in the regulation of mitochondrial cell regeneration..AMPK is an important protein kinase in eukaryotic cells. The role of AMPK in cell energy metabolism is largely dependent on regulating the activity of metabolic enzymes. Our previous study showed that transcutaneous electric acupoint stimulation can effectively reduce the metabolism enzyme and mitochondrion destruction,prolong the working time, delay the skeletal muscle fatigue ,which caused by the excessive exercise.There is speculation that mitochondrial regeneration may be the mechanism of transcutaneous electrical acupoint stimulation in preventing exercise-induced skeletal muscle fatigue.But till now, little is known about the exact role and mechanism of AMPK on the preventing mitochondrial biosynthesis within Exercise-Induced Muscular Fatigue by Transcutanclus Electrical Acupoint Stimulation.This study is to explore the role and molecular mechanism of mitochondrial biogenesis of transcutaneous electric acupoint stimulation in preventing exercise-induced skeletal muscle fatigue,by observating PGC-1α, NRF-1, Tfam, COX,IV gene expression, AMPK phosphorylation, PGC-1α, NRF-1, mtTFA, COX IV protein changes and the COX activity in rat skeletal muscle.

近年来，国内外研究发现AMPK在调控细胞线粒体再生中发挥关键作用。AMPK作为真核生物细胞中一个重要的蛋白激酶，参与细胞的能量代谢过程，其通过调控代谢相关酶起作用。我们前期研究表明经皮穴位电刺激能较好的降低过量运动导致的骨骼肌能量代谢酶和线粒体的破坏，延长肌肉工作的时间，延缓骨骼肌疲劳的发生，故推测促进线粒体再生可能是经皮穴位电刺激防治运动性骨骼肌疲劳的作用机制。但是AMPK是否参与经皮穴位电刺激防治运动性骨骼肌疲劳线粒体生物合成目前还未见报道。本课题通过观察大鼠骨骼肌PGC-1α、NRF-1、mtTFA、COX IV基因表达及骨骼肌磷酸化AMPK蛋白水平、PGC-1α及NRF-1、mtTFA、COX IV蛋白改变及COX活性，探讨经皮穴位电刺激防治运动性骨骼肌疲劳的线粒体生物合成的作用及分子机制。

本课题通过跑台运动方法，制作大鼠运动性骨骼肌疲劳模型。通过观察大鼠腓肠肌能量代谢相关酶的含量；腓肠肌超微结构的变化；腓肠肌AMPK、PGC-1α、NRF-1、mtTFA、COX IV基因表达及AMPK、PGC-1α、NRF-1、mtTFA、COX IV蛋白改变，探讨经皮穴位电刺激（TEAS）防治运动性骨骼肌疲劳的线粒体生物合成的作用及分子机制。得出①TEAS可以减轻大鼠骨骼肌的损伤，降低大鼠血清CK和CK-MM的含量；②TEAS可减少大鼠骨骼肌糖原的消耗，减少乳酸堆积；③TEAS可减少大鼠骨骼肌线粒体的破坏，具有稳定骨骼肌细胞能量代谢的作用；④TEAS可通过促进大鼠骨骼肌AMPK、NRF-1、PGC-1α、mtTFA、COX IV 蛋白和基因表达，参与骨骼肌线粒体再生，从而起到防治运动性骨骼肌疲劳的作用。