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PINK1/Parkin介导的线粒体自噬在运动性骨骼肌损伤中的作用

Role of PINK1/Parkin-mediated Mitophagy in Exercise-induced Skeletal Muscle Damage

  • 摘要:
      目的  探讨大负荷运动对大鼠骨骼肌线粒体自噬蛋白PINK1和Parkin表达的影响,分析PINK1/Parkin途径在运动致骨骼肌线粒体损伤中的作用。
      方法  48只成年雄性Sprague-Dawley大鼠,随机分为安静对照组(C,n=8)和运动组(E,n=40)。其中,E组按时相点划分为0 h、12 h、24 h、48 h、72 h等5组,每组8只。E组大鼠在跑台上进行持续性下坡跑,跑台坡度为-16°,速度为16 m/min,运动时间为90 min。各组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的变化,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,应用Western Blot方法检测骨骼肌PINK1和线粒体中Parkin、LC3的蛋白表达。
      结果  一次大负荷运动后比目鱼肌线粒体出现明显的肿胀以及在肌膜下积聚等超微结构的异常变化,且伴有大量的处于不同成熟阶段的自噬体形成,同时CS的含量明显减少,线粒体自噬蛋白PINK1、Parkin和LC3均出现一过性表达升高(P < 0.05或P < 0.01)。
      结论  一次大负荷运动后,骨骼肌线粒体结构和数量受损,有大量自噬体形成,其致骨骼肌损伤机制可能与PINK1/Parkin途径的活化及继发的过度线粒体自噬有关。

     

    Abstract:
      Objectives  To explore the effect of heavy load exercise on the mitophagy protein expression of PINK1 and Parkin in skeletal muscle of rats, and to analyze the role of PINK1/Parkin pathway in exercise-induced mitochondrial damage in skeletal muscles.
      Methods  Forty-eight male adult SD rats were randomly divided into quiet control group (C group, n=8) and exercise group (E group, n=40).Among them, E group was further divided into 0 h, 12 h, 24 h, 48 h and 72h sub-groups(n=8) according to the time point.Rats in the E group performed a running on a treadmill down a 16° incline at 16m/min for 90 min, 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) was measured by ELISA; Protein expression of skeletal muscle PINK1 and mitochondrial Parkin and LC3 were detected by Western blot.
      Results  After heavy load exercise, the mitochondrial structure appeared the typical abnormal changes such as swelling and accumulating under cell membrane, and formed a large number of mitophagosomes which were in different stages of maturity.The content of CS was significantly decreased, and the protein expression of PINK1, Parkin and LC3 were significantly elevated (P < 0.05 or P < 0.01).
      Conclusion  After a heavy load exercise, the structure and quantity of mitochondria in skeletal muscle were impaired, and a large number of mitophagosomes were formed.The mechanism of skeletal muscle damage caused by heavy load exercise may be related to the activation of PINK1/Parkin pathway and the further initiating overactive mitophagy.

     

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