大负荷运动诱导大鼠骨骼肌损伤对其自噬超微结构及Beclin1和LC3-Ⅱ/Ⅰ的影响  

Effects of high-load exercise induced skeletal muscle injury on autophagy ultrastructure and Beclin1 and LC3-Ⅱ/Ⅰ in rats

作　　者：张欣 王瑞元[2] ZHANG Xin;WANG Rui-yuan(School of Sport and Health,Nanjing Sport Institute,Nanjing 210014;Sport Science College,Beijing Sport University,Beijing 100084,China)

机构地区：[1]南京体育学院运动健康科学学院,江苏南京210014 [2]北京体育大学运动人体科学学院,北京100084

出　　处：《中国应用生理学杂志》2020年第4期296-300,I0001,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金项目(31471133);南京体育学院国家级培育项目(PY201921)。

摘　　要：目的:观察大负荷离心运动对大鼠骨骼肌自噬超微结构及自噬相关蛋白Beclin1和LC3Ⅱ/Ⅰ的影响。方法:48只SD雄性大鼠适应性训练后随机分成对照组(C,n=8)和大负荷离心运动组(E,n=40)。E组于跑台进行90 min下坡跑,运动后0 h、12 h、24 h、48 h和72 h取比目鱼肌,透射电镜观察其自噬体超微结构变化;Western blot检测Beclin1和LC3Ⅱ/Ⅰ蛋白表达;免疫荧光观测LC3的定位及含量变化。结果:E组比目鱼肌自噬体数量在运动后0 h、12 h和24 h均有增加,并伴LC3自噬荧光明显增强(P<0.01),同时运动后48 h自噬荧光仍有显著性升高(P<0.05);Beclin1和LC3Ⅱ/Ⅰ在大负荷离心干预后表达升高(P<0.05),运动后12 h^24 h达到峰值(P<0.01),直至运动后72 h完全恢复。结论:大负荷离心运动可诱导骨骼肌自噬超微结构变化,自噬蛋白表达增强,以上可能是运动损伤的骨骼肌功能下降的原因之一。Objective: To investigate the effects of high-load eccentric exercise on the ultrastructure of autophagy and the autophagy-related proteins Beclin1 and LC3Ⅱ/Ⅰ in rats. Methods: Forty-eight SD male rats were randomly divided into control group( C,n= 8) and high-load eccentric exercise group( E,n = 40) after adaptive training. Group E was run downhill for 90 minutes on the running platform,and soleus muscles were collected at 0,12,24,48 and 72 hours after exercise. Transmission electron microscopy was used to observe the ultrastructural changes of skeletal muscle autophagosomes. Western blot was used to detect the expressions of Beclin1 and LC3Ⅱ/Ⅰ protein. Immunofluorescence was used to observe the localization and content of LC3. Results: The number of soleus muscle autophagosomes in group E was increased at 0,12 and 24 hours after exercise,and LC3 autophagic fluorescence was significantly increased( P<0.01),while autophagic fluorescence at 48 hours after exercise was still increased significantly( P<0.05). Beclin1 and LC3Ⅱ/Ⅰ expression levels were increased after high-load centrifugal intervention( P<0.05),and were peaked at 12 h ~ 24 h after exercise( P<0.01),and fully recovered at 72 h after exercise. Conclusion: High-load eccentric exercise can induce ultrastructural changes in skeletal muscle autophagy and increase the expression of autophagy protein. The peak value appears at 12 hours after exercise. The above may be one of the reasons for the decline in skeletal muscle function caused by sports injury.

关 键 词：骨骼肌损伤 自噬 超微结构 BECLIN1 LC3-Ⅱ/Ⅰ 

分 类 号：G804.2[文化科学—运动人体科学]