针刺对运动性骨骼肌损伤大鼠骨骼肌线粒体自噬的影响

发布时间：2018-03-04 11:00

本文选题：针刺　切入点：大负荷运动　出处：《中国病理生理杂志》2017年11期 　论文类型：期刊论文

【摘要】：目的:通过观察针刺对大负荷运动大鼠骨骼肌线粒体自噬相关蛋白表达的影响,探讨针刺在运动性骨骼肌损伤修复中的作用及其机制。方法:将128只成年雄性Sprague-Dawley大鼠随机分为4组:空白对照(control,C;n=8)组、单纯运动(exercise,E;n=40)组、单纯针刺(acupuncture,A;n=40)组和运动针刺(exercise and acupuncture,EA;n=40)组。其中,E和EA组进行1次下坡跑运动,A组和EA组(运动后即刻)施加针刺处理。后3组根据干预后不同时相又分为0 h、12 h、24 h、48 h和72 h亚组(n=8),分别于对应时点分离比目鱼肌进行检测,使用透射电子显微镜观察骨骼肌线粒体超微结构变化;采用ELISA法检测比目鱼肌线粒体定量酶柠檬酸合成酶(CS)的含量变化;应用Western blot法检测骨骼肌PTEN诱导假定激酶1(PINK1)、parkin和微管相关蛋白1轻链3(LC3)的蛋白表达变化。结果:1次大负荷运动后大鼠比目鱼肌线粒体出现明显肿胀和肌膜下积聚等超微结构异常变化,伴有大量自噬体形成;同时CS的含量明显减少(P0.05);线粒体自噬蛋白PINK1、parkin和LC3均出现一过性的表达升高(P0.05)。运动后针刺明显改善了线粒体超微结构的异常变化,减少自噬溶酶体的出现,同时抑制CS的含量减少,下调PINK1、parkin和LC3在线粒体上的表达(P0.05)。结论:1次大负荷运动后骨骼肌线粒体结构和数量受损,通过激活PINK1/parkin途径诱发线粒体自噬的过度发生。大负荷运动后针刺可以缓解骨骼肌线粒体的损伤,其作用机制可能是通过下调线粒体外膜蛋白PINK1表达,抑制其对下游胞浆蛋白parkin的招募,进而影响LC3与线粒体的结合以抑制线粒体自噬过度激活。
[Abstract]:Objective: to observe the effect of acupuncture on the expression of mitochondrial autophagy related protein in skeletal muscle of rats with heavy exercise. Methods: 128 adult male Sprague-Dawley rats were randomly divided into 4 groups: control group (control group) and exercise exercise group (group 40). Group A and group E were treated with acupuncture for one downhill run, group A and group EA (immediately after exercise). The last three groups were divided into 0 h 12 h, 24 h and 48 h according to the different phases after intervention, and the latter three groups were divided into two groups: group A and group EA (immediately after exercise), and the latter three groups were divided into two groups: group A and group EA (immediately after exercise), and the latter three groups were divided into two groups: group A and group EA (immediately after exercise), and the latter three groups were divided into two groups according to different phases after intervention. The soleus muscle was detected at the corresponding time points. The ultrastructure of mitochondria in skeletal muscle was observed by transmission electron microscope (TEM), and the content of mitochondrial quantitative enzyme citrate synthase (CSS) in soleus muscle was detected by ELISA method. Western blot method was used to detect the changes of protein expression in skeletal muscle induced by PTEN (PINK1) parkin and microtubule-associated protein 1 light chain 3 (LC3). Results the mitochondria of rat soleus muscle showed significant swelling and submuscular accumulation after one heavy exercise. Abnormal changes in microstructure, Meanwhile, the contents of CS decreased significantly, the expression of mitochondrial autophagy protein PINK1 parkin and LC3 increased temporarily. After exercise, acupuncture significantly improved the ultrastructure of mitochondria and reduced the appearance of autophagy lysosomes. At the same time, the contents of CS were inhibited, and the expression of PINK1 parkin and LC3 in mitochondria was down-regulated. Conclusion the structure and number of skeletal muscle mitochondria were damaged after one heavy exercise. The mitochondrial autophagy was induced by activating the PINK1/parkin pathway. Acupuncture after heavy exercise can alleviate the mitochondrial damage of skeletal muscle, and its mechanism may be by down-regulating the expression of mitochondrial outer membrane protein (PINK1). It inhibited the recruitment of downstream cytoplasmic protein parkin, and then inhibited the mitochondrial autophagy hyperactivation by affecting the binding of LC3 to mitochondria.
【作者单位】：成都体育学院运动医学与健康学院;北京体育大学运动人体科学学院;井冈山大学体育学院;
【基金】：国家自然科学基金资助项目(No.31471133) 国家体育总局运动医学重点实验室资助项目(No.CX17B04) 成都体育学院校级科研项目(No.17YJ04);成都体育学院运动医学与健康研究所创新课题(No.CX17C02)
【分类号】：R87

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