miR-708 affords protective efficacy in anoxia/reoxygenation-stimulated cardiomyocytes by blocking the TLR4 signaling via targeting HMGB1

Mol Cell Probes. 2020 Dec:54:101653. doi: 10.1016/j.mcp.2020.101653. Epub 2020 Aug 28.

Abstract

Ischemic heart disease is a proverbial and common cardiovascular disease, and constitutes a leading cause of disability and mortality globally. Myocardial ischemic/reperfusion (MI/R) injury is a highly orchestrated phenomenon that involves the excessive activation of high mobility group box 1 (HMGB1) signaling. In the present study, we sought to investigate the function of miR-708 in MI/R injury due to the predicted binding to HMGB1. Intriguingly, down-regulation of miR-708 and up-regulation of HMGB1 were observed in MI/R rat model and H9c2 cardiomyocytes exposed to hypoxia/reoxygenation (H/R) conditions. Dual luciferase reporter assays substantiated that HMGB1 was a direct target of miR-708. Moreover, miR-708 overexpression suppressed the mRNA and protein expression of HMGB1. Noticeably, elevation of miR-708 antagonized H/R-induced inhibition in cell viability; whilst, increased cell apoptosis evoked by H/R was restrained after miR-708 up-regulation. Simultaneously, miR-708 elevation suppressed H/R exposure-increased lactate dehydrogenase (LDH) release and reactive oxygen species (ROS) generation, but elevated the activity of anti-oxidative stress superoxide dismutase (SOD). Additionally, H/R-increased production of pro-inflammatory cytokine TNF-α and IL-6 was offset following miR-708 overexpression. Moreover, enhancement of miR-708 inhibited H/R-evoked activation of the HMGB1-TLR4-NF-κB pathway by inhibiting the protein levels of HMGB1, TLR4 and p-p65 NF-κB. Specially, restoring this pathway offset the protective effects of miR-708 on H/R-induced cardiomyocyte injury. Together, these data indicate that miR-708 may protect against H/R-induced cardiomyocyte damage by directing targeting HMGB1 signaling, implying a promising therapeutic agent against ischemic heart disease including myocardial infarction.

Keywords: Cardiomyocyte apoptosis; HMGB1-TLR4-NF-κB; Inflammation; Myocardial I/R injury; miR-708.

MeSH terms

  • Animals
  • Base Sequence
  • Cardiotonic Agents / metabolism*
  • Cell Line
  • Cytokines / metabolism
  • Disease Models, Animal
  • HMGB1 Protein / metabolism*
  • Hypoxia / genetics*
  • Inflammation Mediators / metabolism
  • Male
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Myocytes, Cardiac / metabolism*
  • Myocytes, Cardiac / pathology
  • NF-kappa B / metabolism
  • Oxidative Stress
  • Oxygen / metabolism*
  • Rats, Sprague-Dawley
  • Signal Transduction*
  • Toll-Like Receptor 4 / metabolism*
  • Up-Regulation / genetics

Substances

  • Cardiotonic Agents
  • Cytokines
  • HMGB1 Protein
  • Inflammation Mediators
  • MicroRNA708, rat
  • MicroRNAs
  • NF-kappa B
  • Toll-Like Receptor 4
  • Oxygen