Traditional Chinese Medicine manipulation ameliorates oxidative stress injury in skeletal muscle cells model of impairment: A quantitative iTRAQ proteomics study

Research Objectives Rolling manipulation (RM), a myofascial release technique, is the most popular technique in Traditional Chinese Medicine manipulation (TCMM) to treat skeletal muscle injury. The research is to identify potential mechanisms of TCMM alleviating impaired human skeletal muscle cells (HSkMCs) that is based on the changes in related biological processes and pathways after treatment. Design Multiple groups of repeated-measures design. Setting Research laboratory. Participants Normal and injured HSkMCs (ScienCell Research Laboratory, Carlsbad, USA) in a three-dimensional (3-D) culture environment. Interventions Dexamethasone sodium phosphate (DSP) with a concentration of 0.6 to 3.6 mM was used to damage HSKMCs, and the intervention time ranged from 24h to 96h. Intermittent pressure imitating rolling manipulation (IPIRM) was used to dynamically simulate RM via the FX‑5000™ compression system loading with clinical quantitative collection of RM combination of parameters (0-8.8 lbs, 2.0 Hz and 10 min each day for 3 days). Minimal concentration of DSP induced HSKMCs oxidative stress damage and the optimal pressure range of IPIRM were selected to do further proteomic study. Main Outcome Measures Biomarkers (superoxide dismutase activity, malondialdehyde content and creatine kinase activity) for the assessment of lipid peroxidation and oxidative stress status. A global protein profile in normal HSkMCs and injured HSkMCs was obtained via Isobaric tagging for relative and absolute protein quantification (iTRAQ)-based proteomic experiments at baseline and post intervention. Results In the stability of 3-D culture environment, IPIRM whose curve pressure ranging from 0 lbs (lowest pressure) to 2.5 lbs (highest pressure) ameliorated the injured HSkMCs induced by 1.2 mM DSP. Oxidative stress and lipid metabolism signaling pathways were identified as mechanisms underlying the protective effect of TCMM on injured HSkMCs. Western blotting for 4 DEPs (CD36, Hsp27, FABP4 and SelN) confirmed the reliability of iTRAQ. Conclusions IPIRM ranging from 0 to 2.5 lbs pressure downregulated proteins in oxidative stress and lipid metabolism pathways, alleviating excessive oxidative stress and lipid metabolism disorder in injured HSkMCs. Author(s) Disclosures None.