CIRBP protects H9C2 cells against myocardial ischemia through inhibition of NF-κB pathway

Braz J Med Biol Res. 2017 Mar 23;50(4):e5861. doi: 10.1590/1414-431X20175861.


Myocardial ischemia is a major cause of death and remains a disease with extremely deficient clinical therapies and a major problem worldwide. Cold inducible RNA-binding protein (CIRBP) is reported to be involved in multiple pathological processes, including myocardial ischemia. However, the molecular mechanisms of myocardial ischemia remain elusive. Here, we first overexpressed CIRBP by transfection of pc-CIRBP (pcDNA3.1 containing coding sequenced for CIRBP) and silenced CIRBP by transfection of small interfering RNA targeting CIRBP (siCIRBP). pcDNA3.1 and the negative control of siCIRBP (siNC) were transfected into H9C2 cells to act as controls. We then constructed a cell model of myocardial ischemia through culturing cells in serum-free medium with hypoxia in H9C2 cells. Subsequently, AlamarBlue assay, flow cytometry and western blot analysis were used, respectively, to assess cell viability, reactive oxygen species (ROS) level and apoptosis, and expression levels of IκBα, p65 and Bcl-3. We demonstrated that CIRBP overexpression promoted cell proliferation (P<0.001), inhibited cell apoptosis (P<0.05), reduced ROS level (P<0.001), down-regulated phosphorylated levels of IκBα and p65 (P<0.01 or P<0.001), and up-regulated expression of Bcl-3 (P<0.001) in H9C2 cells with myocardial ischemia. The influence of CIRBP knockdown yielded opposite results. Our study revealed that CIRBP could protect H9C2 cells against myocardial ischemia through inhibition of NF-κB pathway.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Blotting, Western
  • Cell Proliferation / drug effects
  • Cell Survival
  • Cells, Cultured
  • Flow Cytometry
  • Gene Expression Regulation
  • Myocardial Ischemia / metabolism*
  • Myocardial Ischemia / prevention & control*
  • NF-kappa B / antagonists & inhibitors*
  • NF-kappa B / metabolism
  • Protective Agents / pharmacology*
  • RNA, Small Interfering
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism
  • RNA-Binding Proteins / pharmacology*
  • Rats
  • Reactive Oxygen Species / analysis
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Time Factors
  • Transfection / methods


  • CIRBP protein, human
  • NF-kappa B
  • Protective Agents
  • RNA, Small Interfering
  • RNA-Binding Proteins
  • Reactive Oxygen Species