Targeting MIAT reduces apoptosis of cardiomyocytes after ischemia/reperfusion injury

Bioengineered. 2019 Dec;10(1):121-132. doi: 10.1080/21655979.2019.1605812.

Abstract

This study aims to investigate the role of targeting lncRNA myocardial infarction-associated transcript (MIAT) in protection against hypoxia/reoxygenation (H/R) injury in H9c2 cells in vitro and myocardial ischemia/reperfusion (I/R) injury in vivo by regulating expression of NF-kB and p53 upregulated modulator of apoptosis (PUMA). H9C2 cells were infected with lentivirus expressing the short-hairpin RNA direct against human MIAT gene (Lv-MIAT shRNA) or lentivirus expressing scrambled control (Lv-NC shRNA) or PUMA siRNA or p65 siRNA or their control siRNA respectively. Then the H9c2 cells were infected with Lv-shRNA to 2 hours of hypoxia (H) and 24 hour of reoxygenation (R). 100 ul of Lv-MIAT shRNA (1 × 108 PFU) or Lv-NC shRNA was transfected into mouse hearts, then the hearts were subjected to I/R (1h/72 h). We discovered targeting MIAT remarkably enhanced H9c2 cell viability, decreased H/R-induced cell apoptosis and LDH leakage and significantly decreased I/R-induced myocardial infarct size, reduced myocardial apoptosis and enhanced the heart function. Targeting MIAT downregulated p65 nuclear translocation, NF-κB activity and anti-apoptotic protein cleaved-caspase-3, Bax, and upregulated anti-apoptotic protein Bcl-2 induced by H/R or I/R. Our study suggests that targeting MIAT may protect against H9c2 cardiomyoblasts H/R injury or myocardial I/R injury via inhibition of cell apoptosis, mediated by NF-κB and PUMA signal pathway.

Keywords: Hypoxia/reoxygenation; apoptosis; ischaemia-reperfusion; lncRNA myocardial infarction-associated transcript; nuclear factor kappa B; p53 upregulated modulator of apoptosis.

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Apoptosis Regulatory Proteins / antagonists & inhibitors
  • Apoptosis Regulatory Proteins / genetics*
  • Apoptosis Regulatory Proteins / metabolism
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Cell Hypoxia
  • Cell Line
  • Cell Survival
  • Gene Expression Regulation
  • Humans
  • L-Lactate Dehydrogenase / metabolism
  • Mice
  • Myocardial Reperfusion Injury / genetics*
  • Myocardial Reperfusion Injury / metabolism
  • Myocardial Reperfusion Injury / pathology
  • Myocardial Reperfusion Injury / prevention & control
  • Myocardium / metabolism*
  • Myocardium / pathology
  • Myocytes, Cardiac / metabolism*
  • Myocytes, Cardiac / pathology
  • Protein Transport
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Long Noncoding / antagonists & inhibitors
  • RNA, Long Noncoding / genetics*
  • RNA, Long Noncoding / metabolism
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Rats
  • Signal Transduction
  • Transcription Factor RelA / antagonists & inhibitors
  • Transcription Factor RelA / genetics*
  • Transcription Factor RelA / metabolism
  • Tumor Suppressor Proteins / antagonists & inhibitors
  • Tumor Suppressor Proteins / genetics*
  • Tumor Suppressor Proteins / metabolism
  • bcl-2-Associated X Protein / genetics
  • bcl-2-Associated X Protein / metabolism

Substances

  • Apoptosis Regulatory Proteins
  • Bax protein, mouse
  • Miat long non-coding RNA
  • PUMA protein, mouse
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Long Noncoding
  • RNA, Small Interfering
  • Rela protein, mouse
  • Transcription Factor RelA
  • Tumor Suppressor Proteins
  • bcl-2-Associated X Protein
  • Bcl2 protein, mouse
  • L-Lactate Dehydrogenase
  • Casp3 protein, mouse
  • Caspase 3