Leucine attenuates muscle atrophy and autophagosome formation by activating PI3K/AKT/mTOR signaling pathway in rotator cuff tears
Rotator cuff tears (RCTs), the most common tendon injury, are always accompanied by muscle atrophy, which is characterized by excessive protein degradation. Autophagy–lysosome systems are the crucial proteolytic pathways and are activated in atrophying muscle. Thus, inhibition of the autophagy–lysosome pathway might be an alternative way to minimize skeletal muscle atrophy. In this present study, combined with a tendon transection-induced rat model of massive rotator cuff tears, HE staining and transmission electron microscopy methods, we found leucine supplementation effectively prevented muscle atrophy, muscle injury and autophagosome formation. Utilizing immunoblotting, we discovered that leucine supplementation is able to inhibit the rise in autophagy-related protein expression (including LC3, Atrogin-1, MuRF1, Bnip3 and FoxO3) driven by tendon transection. The PI3K/AKT/mTOR pathway that was essential in autophagosome formation and autophagy was blocked in atrophying muscle and reactivated in the presence of leucine. Once administrated with LY294002 (PI3K inhibitor) and Rapamycin (mTOR inhibitor), leucine mediated by the anti-atrophic effects was nearly blunted. These results suggest that leucine potentially attenuates tendon transection-induced muscle atrophy through autophagy inhibition via activating the PI3K/AKT/mTOR pathway.
KeywordsRotator cuff tears Skeletal muscle atrophy Leucine PI3K/AKT/mTOR pathway Autophagosome
Dr. Rongzong Zheng (MD), the director of orthopedics, sports medicine and joint surgery sub-specialty conceived,designed and launched this research.
Dr. Shuming Huang (MD) performed the experiments and analyzed data, taking the responsibility of the chief instructor of orthopedics trauma sub-specialty, and shares the privilege of co-first authors and corresponding author.
Dr. Junkun Zhu (MD), the director of orthopedics rehabilitation sub-specialty, submitted the proposal and composed the manuscript and shares the privilege of co-first authors.
Wei Lin, Huan Xu and Xiang Zheng participated and collected the experiment data as well as completing statistics analysis.
This study is supported by Zhejiang Provincial Natural Science Foundation (grant number: Y16H060008, PR China) and Medical and Health Research Program of Zhejiang Province (grant number: 2012KYB249, 2017KY725, PR China).
Compliance with ethical standards
All animal care procedures were approved by laboratory animals of the Fifth Affiliated Hospital of Wenzhou Medical College in this study.
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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