针刺对运动性骨骼肌损伤大鼠细胞自噬及Omi/HtrA2通路的影响  

Effects of Acupuncture on Autophagy and Omi/HtrA2 Pathway in Rats with Exercise-induced Skeletal Muscle Injury

作　　者：张欣 李俊平[2] 王瑞元[2] Zhang Xin;Li Junping;Wang Ruiyuan(Nanjing Sport Institute,Nanjing 210014,China;Beijing Sport University,Beijing 100084,China)

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

出　　处：《中国运动医学杂志》2022年第8期625-632,共8页Chinese Journal of Sports Medicine

基　　金：国家自然科学基金项目(31471133);江苏省高等学校自然科学面上项目(21KJB340001)。

摘　　要：目的:探究针刺对运动性骨骼肌损伤大鼠细胞自噬及Omi/HtrA2通路的影响,为探讨针刺改善运动性骨骼肌损伤的机理提供依据。方法:将128只8周龄SD雄性大鼠随机分为对照组(C组,n=8)、针刺组(A组,n=40)、运动组(E组,n=40)、运动针刺组(EA组,n=40)。E组进行一次持续性下坡跑,跑台坡度为-16°,速度为16 m/min,运动时间为90 min;A组斜刺大鼠比目鱼肌,进针角度约30°,留针2 min;EA组在运动后斜刺比目鱼肌留针干预。分别于运动、针刺或运动针刺干预后0 h、12 h、24 h、48 h和72 h取材。透射电镜观察比目鱼肌细胞内自噬体超微结构变化;Western Blot法检测比目鱼肌丝氨酸蛋白酶Omi/HtrA2、HS1相关蛋白X-1(Hax-1)和自噬蛋白Beclin1的蛋白表达。结果:E组运动后比目鱼肌细胞内出现明显的肌小节撕裂以及细胞器肿胀、异常聚集等超微结构变化,并伴有大量的自噬体形成,同时Omi/HtrA2和Beclin1的蛋白表达量在0 h、12 h、24 h、48 h时间点均较C组升高(P<0.05或P<0.01),前者在12 h时间点达到最高,后者在24 h时间点达到最高,两者均在72 h时间点恢复;Hax-1蛋白表达在0 h、12 h、24 h、48 h时间点较C组出现降低(P<0.05或P<0.01),在12 h时间点达到最低,72 h时间点恢复。A组各目标蛋白表达量在各个时间点与C组相比均无显著性差异。EA组运动加针刺干预后比目鱼肌细胞内肌小节撕裂以及细胞器肿胀、异常聚集等超微结构变化相对明显,自噬体形成不活跃,同时Omi/HtrA2和Beclin1的蛋白表达在0 h、12 h、24 h、48 h时间点较C组有升高趋势(P<0.05或P<0.01),最高值均出现在12 h时间点,Hax-1蛋白表达在0 h、12 h、24 h、48 h时间点较C组出现降低(P<0.05或P<0.01),最低值出现在12 h时间点。EA组相较于A组,除Hax-1蛋白表达持续降低至72 h时间点(P<0.05)外,其他各目标蛋白变化与EA组相较于C组的结果相同。EA组相较于E组,Omi/HtrA2和Beclin1的蛋白表达在0 h、12 h�Objective To explore the effect of acupuncture on autophagy and Omi/HtrA2 pathway of rats with exercise-induced skeletal muscle injury,so as to provide evidences for the mechanism of acupuncture improving exercise-induced skeletal muscle injury. Methods One hundred and twenty-eight 8-week-old male Sprague-Dawley rats were randomly divided into a control group(C,n=8),an acupuncture group(A,n=40),an exercise group(E,n=40) and an exercise-acupuncture group(EA,n=40).Group E underwent a continuous 90-minute downhill running on a slope of-16°,at a speed of 16 m/min. The soleus muscles of group A were obliquely punctured at an angle of 30° for 2 minutes,while those of group EA were punctured in the same way after exercise. Samples were taken at 0 h,12 h,24 h,48 h and 72 h after exercise,acupuncture or exercise & acupuncture. Ultrastructural changes of autophagosomes in soleus muscle cells were observed using the transmission electron microscope.The protein expressions of Omi/HtrA2,HS1-associated protein X-1(Hax-1) and autophagic protein Beclin1 were detected using Western blotting. Results In Group E,significant ultrastructural changes such as sarcomere laceration,organelle swelling and abnormal aggregation were found in soleus muscle cells after exercise,accompanied by the formation of a large number of autophagosomes. The protein expression of Omi/HtrA2 and Beclin1 of group E increased at 0 h,12 h,24 h,48 h compared with group C(P<0.05 or P<0.01),with the former and latter reaching the highest at 12 h and 24 h,and both recovering at 72 h. The expression of Hax-1 protein of group E decreased at 0 h,12 h,24 h and 48h compared with group C(P<0.05 or P<0.01),reaching the lowest at 12 h and recovering at 72 h.However,there was no significant difference between group A and C in the protein expressions. After the intervention,the sarcomere laceration,organelles swelling and abnormal aggregation in soleus muscle cells of group EA were relatively obvious,but the autophagy was not active,with the protein expression of Omi/Htr

关 键 词：骨骼肌 自噬 OMI/HTRA2 针刺 

分 类 号：R245[医药卫生—针灸推拿学]