miR-520d suppresses rapid pacing-induced apoptosis of atrial myocytes through mediation of ADAM10

J Mol Histol. 2021 Apr;52(2):207-217. doi: 10.1007/s10735-020-09938-w. Epub 2021 Feb 5.

Abstract

MicroRNAs (miRNAs) play a key role in various pathological processes like atrial fibrillation (AF). However, the mechanisms remain unclear. Herein, this study was undertaken to probe the roles of ADAM10 and its targeting miR-520d in rapid pacing-induced apoptosis in atrial myocytes. In this study, the atrial myocytes grew adherently with irregular morphology. Immunofluorescence showed that more than 90% of atrial myocytes were α-sarcomeric actin (α-SCA) positive, indicating that the primary cells were positive for α-SCA staining and atrial myocytes were successfully isolated. The pacing atrial myocyte model was established after rapid pacing stimulation and we found the rapid pacing stimulation caused elevated ADAM10 and suppressed miR-520d. CCK-8 assay was applied for evaluation of cell viability, TUNEL staining for assessment of cell apoptosis and dual-luciferase reporter gene assay for verification of the targeting relationship between miR-520d and ADAM10. Overexpression of miR-520d or silencing of ADAM10 could enhance cell viability and reduce cell apoptosis in the rapid pacing-induced atrial myocytes. ADAM10 was a target gene of miR-520d. MiR-520d negatively targeted ADAM10, thereby promoting cell viability and inhibiting apoptosis in rapid pacing atrial myocyte model. In summary, miR-520d enhances atrial myocyte viability and inhibits cell apoptosis in rapid pacing-induced AF mouse model through negative mediation of ADAM10.

Keywords: ADAM10; Atrial myocyte; Cell apoptosis; Cell viability; Rapid pacing; miR-520d.

MeSH terms

  • ADAM10 Protein / genetics
  • ADAM10 Protein / metabolism*
  • Amyloid Precursor Protein Secretases / genetics
  • Amyloid Precursor Protein Secretases / metabolism*
  • Animals
  • Apoptosis / genetics
  • Apoptosis / physiology*
  • Cell Proliferation / genetics
  • Cell Proliferation / physiology
  • Cell Survival / genetics
  • Cell Survival / physiology
  • Fluorescent Antibody Technique
  • In Situ Nick-End Labeling
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Myocytes, Cardiac / metabolism*
  • Myocytes, Cardiac / physiology*
  • Signal Transduction / genetics
  • Signal Transduction / physiology

Substances

  • Membrane Proteins
  • MicroRNAs
  • Amyloid Precursor Protein Secretases
  • ADAM10 Protein
  • Adam10 protein, mouse