Optimizing mechanical stretching protocols for hypertrophic and anti-apoptotic responses in cardiomyocyte-like H9C2 cells

Mol Biol Rep. 2021 Jan;48(1):645-655. doi: 10.1007/s11033-020-06112-z. Epub 2021 Jan 4.

Abstract

Cardiomyocytes possess the ability to respond to mechanical stimuli by reprogramming their gene expression. This study investigated the effects of different loading protocols on signaling and expression responses of myogenic, anabolic, inflammatory, atrophy and pro-apoptotic genes in cardiomyocyte-like H9C2 cells. Differentiated H9C2 cells underwent various stretching protocols by altering their elongation, frequency and duration, utilizing an in vitro cell tension system. The loading-induced expression changes of MyoD, Myogenin, MRF4, IGF-1 isoforms, Atrogin-1, Foxo1, Fuca and IL-6 were measured by Real Time-PCR. The stretching-induced activation of Akt and Erk 1/2 was also evaluated by Western blot analysis. Low strain (2.7% elongation), low frequency (0.25 Hz) and intermediate duration (12 h) stretching protocol was overall the most effective in inducing beneficial responses, i.e., protein synthesis along with the suppression of apoptosis, inflammation and atrophy, in the differentiated cardiomyocytes. These findings demonstrated that varying the characteristics of mechanical loading applied on H9C2 cells in vitro can regulate their anabolic/survival program.

Keywords: Cardiomyocytes; Cellular mechanotransduction; H9C2; Mechanical stretch.

MeSH terms

  • Animals
  • Apoptosis / genetics*
  • Cell Death / genetics
  • Cell Line
  • Cell Survival / genetics
  • Cellular Reprogramming / genetics*
  • Gene Expression Regulation, Developmental / genetics
  • Humans
  • Hypertrophy / genetics*
  • Hypertrophy / pathology
  • Inflammation / genetics
  • Inflammation / metabolism
  • Inflammation / pathology
  • Insulin-Like Growth Factor I / genetics
  • MAP Kinase Signaling System / genetics
  • Mechanotransduction, Cellular / genetics*
  • Muscle Proteins / genetics
  • MyoD Protein / genetics
  • Myocytes, Cardiac / metabolism*
  • Myocytes, Cardiac / pathology
  • Myogenic Regulatory Factors / genetics
  • Myogenin / genetics
  • Rats
  • SKP Cullin F-Box Protein Ligases / genetics

Substances

  • Muscle Proteins
  • MyoD Protein
  • Myogenic Regulatory Factors
  • Myogenin
  • myogenic factor 6
  • Insulin-Like Growth Factor I
  • Fbxo32 protein, rat
  • SKP Cullin F-Box Protein Ligases