Simultaneous miRNA and mRNA Transcriptome Profiling of Differentiating Equine Satellite Cells Treated with Gamma-Oryzanol and Exposed to Hydrogen Peroxide

Nutrients. 2018 Dec 2;10(12):1871. doi: 10.3390/nu10121871.

Abstract

Gamma-oryzanol (GO) is a popular supplement for performance horses, dogs, and humans. Previous studies indicated that GO supplementation decreases creatine kinase activity and lactate level after exercise and may affect oxidative stress in Thoroughbred horses. GO may change genes expression in equine satellite cells (ESC). The purpose of this study was to evaluate the effect of GO on miRNA, gene expression, oxidative stress, and cell damage and viability in differentiating ESC pretreated with hydrogen peroxide (H₂O₂). ESCs were obtained from a young horse's skeletal muscle. ESCs were pre-incubated with GO (24 h) and then exposed to H₂O₂ for one hour. For the microRNA and gene expression assessment, the microarray technique was used. Identified miRNAs and genes were validated using real time-quantitative polymerase chain reaction. Several tests related to cell viability, cell damage, and oxidative stress were performed. The microarray analysis revealed differences in 17 miRNAs and 202 genes between GO-treated and control ESC. The tests related to apoptosis, cell viability, and oxidative stress showed that GO affects these processes to varying degrees. Our results suggest that GO can change miRNA and gene expression and may impact the processes involved in tissue repairing after an injury.

Keywords: apoptosis; equine satellite cells; gamma-oryzanol; hydrogen peroxide; miRNA; muscle injuries; oxidative stress; target genes.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cell Differentiation / drug effects*
  • Cell Differentiation / genetics
  • Cell Proliferation / genetics
  • Cell Survival / drug effects
  • Cells, Cultured
  • Gene Expression / drug effects
  • Gene Expression Profiling / methods
  • Gene Expression Profiling / veterinary*
  • Horses*
  • Hydrogen Peroxide / pharmacology*
  • Male
  • MicroRNAs / analysis
  • Oxidative Stress / drug effects
  • Phenylpropionates / pharmacology*
  • RNA, Messenger / analysis
  • Satellite Cells, Skeletal Muscle / drug effects
  • Satellite Cells, Skeletal Muscle / physiology*
  • Tissue Array Analysis / methods
  • Tissue Array Analysis / veterinary

Substances

  • MicroRNAs
  • Phenylpropionates
  • RNA, Messenger
  • Hydrogen Peroxide
  • gamma-oryzanol