针刺对大负荷运动大鼠骨骼肌线粒体结构和功能的影响

关键词：大负荷运动  针刺  骨骼肌  线粒体  呼吸链复合体

Keywords:heavy load exercise  acupuncture  skeletal muscle  mitochondrion  respiratory chain complex

摘要：

      摘要 目的：研究针刺对大负荷运动大鼠骨骼肌线粒体结构与功能的影响，探讨针刺干预对运动性骨骼肌损伤的恢复和预防的作用。 方法：128只成年雄性SD大鼠，随机分为4组：空白对照组（C组,n=8）、单纯运动组（E组,n=40）、单纯针刺组（A组,n=40）和运动针刺组（EA组,n=40）。其中，E和EA组进行一次下坡跑运动，A组和EA组在运动后即刻施加针刺处理。各组根据干预后不同时相又分为0h、12h、24h、48h和72h组（n=8），分别于对应时间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方法检测各组大鼠比目鱼肌线粒体定量酶CS的含量以及线粒体呼吸链复合体Ⅱ、Ⅳ的活性，应用Western Blot方法检测骨骼肌COXⅠ的蛋白表达。 结果：单纯针刺后骨骼肌COXⅠ蛋白表达上调。大负荷运动后比目鱼肌线粒体出现明显肿胀、肌膜下积聚等超微结构异常变化，且伴有大量自噬体形成，同时柠檬酸合成酶（CS）的含量明显减少，呼吸链复合体Ⅱ、Ⅳ活性以及COXⅠ蛋白表达出现一过性的下调（P<0.05）。与运动组相比，运动针刺后明显改善了大负荷运动所致线粒体超微结构的异常变化，有自噬溶酶体的出现，CS含量有所增加，呼吸链复合体Ⅱ活性及COXⅠ蛋白表达上调。 结论：针刺促进了大负荷运动后骨骼肌线粒体结构的恢复，有效增加了运动后线粒体的数量，减轻了大负荷运动对线粒体功能的损害。

Abstract:

      Abstract Objective: To study the influence of acupuncture on mitochondrial structure and function of skeletal muscle in rats with heavy load exercise, and to explore the effect of acupuncture on the recovery and prevention of exercise-induced skeletal muscle damage. Method: One hundred and twenty-eight male adult SD rats were randomly divided into four groups: control group (C, n=8), exercise group (E, n=40), acupuncture group (A, n=40), exercise and acupuncture group (EA, n=40). Rats in E and EA groups were subjected to an eccentric exercise, and those in A and EA groups were applied acupuncture treatment immediately after exercise. Each group was further divided into 0h，12h，24h，48h and 72h sub-groups(n=8), and at each time point the soleus muscle was collected under anesthesia. Mitochondrial ultrastructural changes in skeletal muscle were observed by a transmission electron microscope. The content of quantitative enzyme citrate synthase(CS) and the activities of mitochondrial respiratory chain complex Ⅱand Ⅳwere measured by ELISA. Protein expression of skeletal muscle COXⅠwas determined by western blot. Result: The protein expression of COXⅠin skeletal muscle was up-regulated after acupuncture alone. After the heavy load exercise, the mitochondria structure appeared the typical abnormal changes such as swelling and accumulating under cell membrane, and formed a large number of mitophagosomes. The content of CS, the activities of respiratory chain complex Ⅱ and Ⅳ and the protein expression of COXⅠwere significantly decreased (P<0.05). Compared with the exercise group, acupuncture intervention significantly promoted the recovery of mitochondrial ultrastructure, induced the formation of mitophagolysosome, alleviated the reduction of CS content, increased the activity of respiratory chain complex Ⅱ and the expression of COXⅠin skeletal muscle. Conclusion: Acupuncture can promote the recovery of mitochondrial structure and effectively increase the number of mitochondria after heavy load exercise, thereby alleviating the damage of mitochondrial function within rat skeletal muscle.