Intermittent pressure imitating rolling manipulation ameliorates injury in skeletal muscle cells through oxidative stress and lipid metabolism signalling pathways

Gene. 2021 Apr 30:778:145460. doi: 10.1016/j.gene.2021.145460. Epub 2021 Jan 27.

Abstract

Background: Traditional Chinese medicine manipulation (TCMM) is often used to treat human skeletal muscle injury, but its mechanism remains unclear due to difficulty standardizing and quantifying manipulation parameters.

Methods: Here, dexamethasone sodium phosphate (DSP) was utilized to induce human skeletal muscle cell (HSkMC) impairments. Cells in a three-dimensional environment were divided into the control normal group (CNG), control injured group (CIG) and rolling manipulation group (RMG). The RMG was exposed to intermittent pressure imitating rolling manipulation (IPIRM) of TCMM via the FX‑5000™ compression system. Skeletal muscle damage was assessed via the cell proliferation rate, superoxide dismutase (SOD) activity, malondialdehyde (MDA) content and creatine kinase (CK) activity. Isobaric tagging for relative and absolute protein quantification (iTRAQ) and bioinformatic analysis were used to evaluate differentially expressed proteins (DEPs).

Results: Higher-pressure IPIRM ameliorated the skeletal muscle cell injury induced by 1.2 mM DSP. Thirteen common DEPs after IPIRM were selected. Key biological processes, molecular functions, cellular components, and pathways were identified as mechanisms underlying the protective effect of TCMM against skeletal muscle damage. Some processes (response to oxidative stress, response to wounding, response to stress and lipid metabolism signalling pathways) were related to skeletal muscle cell injury. Western blotting for 4 DEPs confirmed the reliability of iTRAQ.

Conclusions: Higher-pressure IPIRM downregulated the CD36, Hsp27 and FABP4 proteins in oxidative stress and lipid metabolism pathways, alleviating excessive oxidative stress and lipid metabolism disorder in injured HSkMCs. The techniques used in this study might provide novel insights into the mechanism of TCMM.

Keywords: Lipid metabolism; Oxidative stress; Proteomics; Rolling manipulation; Skeletal muscle cell injury; iTRAQ.

MeSH terms

  • Biomechanical Phenomena
  • CD36 Antigens / metabolism*
  • Cell Culture Techniques
  • Cells, Cultured
  • Dexamethasone / adverse effects
  • Dexamethasone / analogs & derivatives*
  • Down-Regulation
  • Fatty Acid-Binding Proteins / metabolism*
  • Heat-Shock Proteins / metabolism*
  • Humans
  • Lipid Metabolism / drug effects
  • Medicine, Chinese Traditional
  • Models, Biological
  • Molecular Chaperones / metabolism*
  • Muscle Fibers, Skeletal / cytology*
  • Muscle Fibers, Skeletal / drug effects
  • Muscle Fibers, Skeletal / metabolism
  • Musculoskeletal Manipulations / methods*
  • Oxidative Stress / drug effects
  • Proteomics
  • Signal Transduction

Substances

  • CD36 Antigens
  • CD36 protein, human
  • FABP4 protein, human
  • Fatty Acid-Binding Proteins
  • HSPB1 protein, human
  • Heat-Shock Proteins
  • Molecular Chaperones
  • dexamethasone 21-phosphate
  • Dexamethasone