Umbilical cord mesenchymal stem cell-conditioned media prevent muscle atrophy by suppressing muscle atrophy-related proteins and ROS generation

In Vitro Cell Dev Biol Anim. 2016 Jan;52(1):68-76. doi: 10.1007/s11626-015-9948-1. Epub 2015 Sep 15.

Abstract

The therapeutic potential of mesenchymal stem cell-conditioned medium (MSC-CM) has been reported with various types of disease models. Here, we examine the therapeutic effect of umbilical cord MSC-CM (UCMSC-CM) on muscle-related disease, using a dexamethasone (Dex)-induced muscle atrophy in vitro model. The expressions of muscle atrophy-related proteins (MuRF-1 and MAFbx) and muscle-specific proteins (desmin and myogenin) were evaluated by Western blot analysis. The level of production of reactive oxygen species (ROS) was determined using a 2',7'-dichlorofluorescein diacetate (DCFDA) dye assay. The expression of antioxidant enzymes (copper/zinc-superoxide dismutase (Cu/Zn-SOD), manganese superoxide dismutase (MnSOD), glutathione peroxidase-1 (GPx-1), and catalase (CAT)) was verified by reverse transcription polymerase chain reaction (RT-PCR). When L6 cells were exposed to Dex, the expression of muscle atrophy-related proteins was increased by 50-70%, and the expression of muscle-specific proteins was in turn decreased by 23-40%. Conversely, when the L6 cells were co-treated with UCMSC-CM and Dex, the expression of muscle atrophy-related proteins was reduced in a UCMSC-CM dose-dependent manner and the expression of muscle-specific proteins was restored to near-normal levels. Moreover, ROS generation was effectively suppressed and the expression of antioxidant enzymes was recovered to a normal degree. These data imply that UCMSC-CM clearly has the potential to prevent muscle atrophy. Thus, our present study offers fundamental data on the potential treatment of muscle-related disease using UCMSC-CM.

Keywords: Dexamethasone; L6 skeletal muscle cell; Muscle atrophy; Umbilical cord mesenchymal stem cell (UCMSC); Umbilical cord mesenchymal stem cell-conditioned medium (UCMSC-CM).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Catalase / biosynthesis
  • Cell Proliferation / genetics
  • Culture Media, Conditioned / pharmacology
  • Desmin / biosynthesis*
  • Desmin / genetics
  • Dexamethasone / toxicity
  • Disease Models, Animal
  • Gene Expression Regulation, Developmental / drug effects
  • Glutathione Peroxidase / biosynthesis
  • Glutathione Peroxidase GPX1
  • Humans
  • Mesenchymal Stem Cell Transplantation*
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / drug effects
  • Muscle Proteins / biosynthesis
  • Muscle Proteins / genetics
  • Muscular Atrophy / chemically induced
  • Muscular Atrophy / genetics
  • Muscular Atrophy / pathology
  • Muscular Atrophy / therapy*
  • Myogenin / biosynthesis*
  • Myogenin / genetics
  • Rats
  • Reactive Oxygen Species / metabolism*
  • SKP Cullin F-Box Protein Ligases / biosynthesis
  • SKP Cullin F-Box Protein Ligases / genetics
  • Superoxide Dismutase / biosynthesis
  • Tripartite Motif Proteins
  • Ubiquitin-Protein Ligases / biosynthesis
  • Ubiquitin-Protein Ligases / genetics
  • Umbilical Cord / cytology
  • Umbilical Cord / drug effects
  • Umbilical Cord / transplantation

Substances

  • Culture Media, Conditioned
  • Desmin
  • Muscle Proteins
  • Myogenin
  • Reactive Oxygen Species
  • Trim55 protein, rat
  • Tripartite Motif Proteins
  • Dexamethasone
  • Catalase
  • Glutathione Peroxidase
  • Superoxide Dismutase
  • superoxide dismutase 2
  • Fbxo32 protein, rat
  • SKP Cullin F-Box Protein Ligases
  • Ubiquitin-Protein Ligases
  • Glutathione Peroxidase GPX1