Yonghui Yu: Beijing Advanced Innovation Center for Food Nutrition and Human Health, China-Canada Joint Lab of Food Nutrition and Health (Beijing), Beijing Engineering and Technology Research Center of Food Additives, Beijing Technology and Business University, Beijing, China.
Longlong Yang: Burn Institute, the First Affiliated Hospital of PLA General Hospital, Beijing, China.
Shaofang Han: Burn Institute, the First Affiliated Hospital of PLA General Hospital, Beijing, China.
Yushou Wu: Burn Institute, the First Affiliated Hospital of PLA General Hospital, Beijing, China.
Lingying Liu: Burn Institute, the First Affiliated Hospital of PLA General Hospital, Beijing, China.
Yang Chang: Burn Institute, the First Affiliated Hospital of PLA General Hospital, Beijing, China.
Xiaoteng Wang: Burn Institute, the First Affiliated Hospital of PLA General Hospital, Beijing, China.
Jiake Chai: Burn Institute, the First Affiliated Hospital of PLA General Hospital, Beijing, China.
INTRODUCTION: Cell autophagy is an important material recycling process and is involved in regulating many vital activities under both physiological and pathological conditions. However, the mechanism of autophagy regulating burn-induced skeletal muscle wasting still needs to be elucidated. METHODS: The rat burn model with 30% total body surface area and L6 cell line were used in this study. An immunofluorescence assay was used to detect autophagic levels. MicroRNA array and real-time PCR were employed to measure miR-190b levels, and its influence on PH domain and leucine-rich repeat protein phosphatase 1 (PHLPP1) protein translation was estimated using luciferase reporter assay. The expression levels of autophagy-related proteins were analyzed by Western blot. Skeletal muscle wasting was evaluated by the ratio of tibias anterior muscle weight to body weight. RESULTS: Our study demonstrates that burn injury promotes expression of the autophagy-related proteins light chain 3 (LC3) and Beclin-1, suppresses expression of Akt and Forkhead box O (FoxO) 3a protein phosphorylation, and increases PHLPP1 protein level which is required for Akt dephosphorylation. miR-190b, the regulator of PHLPP1 protein translation, also significantly decreases after burn injury. Ectopic expression of miR-190b in L6 myoblast cell downregulates PHLPP1 protein expression, elevates Akt and FoxO3a phosphorylation, and subsequently reduces cell autophagy. Finally, suppressing autophagy with 3-methyladenine represses the protein expression of LC3 and Beclin-1 and mitigates burn-induced skeletal muscle wasting. CONCLUSION: Burn injury induced skeletal muscle cell autophagy and subsequently resulted in skeletal muscle wasting via regulating miR-190b/PHLPP1/Akt/FoxO3a signaling pathway.
