Downhill Running Acutely Elicits Mitophagy in Rat Soleus Muscle

Purpose To investigate the effects of downhill treadmill running on mitochondrial structure/function and expression levels of mitophagy-related proteins in rat skeletal muscle. Methods A total of 48 male adult Sprague-Dawley rats were randomly divided into a control group (C, n=8) and an exercise group (E, n=40). Rats in the E group were exercised on a treadmill down a 16° decline at 16m/min for 90min, and were further divided into 0h(E0), 12h(E12), 24h(E24), 48h(E48) and 72h(E72) post-exercise sub-groups (n=8 each). At each time point the soleus muscle was collected under full 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 II and IV were measured by enzyme-linked immunosorbent assay. Protein expression of skeletal muscle cytochrome c oxidase subunit 1 (COX1), PTEN-induced putative kinase 1 (PINK1) and mitochondrial Parkin, microtubule-associated protein 1 light chain 3 (LC3) were determined by Western blot. Mitochondrial co-localizations with Parkin, ubiquitin (Ub), p62/Sequestosome 1 (p62) and LC3 were measured by the immunofluorescence double labeling technique. Results After downhill treadmill running, the skeletal muscle mitochondrial structure changed dramatically and a large amount of mitophagosomes were observed; the CS content and Complex II activity were significantly lower (p

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