Differential-display polymerase chain reaction identifies nicotinamide adenine dinucleotide-ubiquinone oxidoreductase as an ischemia/reperfusion-regulated gene in cardiomyocytes

Chest. 2004 Jan;125(1):228-35. doi: 10.1378/chest.125.1.228.

Abstract

Objective: Cardiac ischemia/reperfusion-induced oxidative damage often occurs in mitochondria. We identified differentially expressed genes in the canine heart after global cardiac ischemia/reperfusion injury was induced during cardiopulmonary bypass (CPB).

Methods: Differential-display polymerase chain reaction (ddPCR) was performed on cardiac tissue from canine hearts with or without global cardiac ischemia/reperfusion injury induced during CPB. Ischemia/reperfusion-associated mitochondrial injury was investigated at the protein level using various cardioplegic solutions and Western blot analysis.

Results: A mitochondrial protein nicotinamide adenine dinucleotide (NADH):ubiquinone oxidoreductase gene was identified on ddPCR. The NADH:ubiquinone oxidoreductase gene was up-regulated in canine hearts after 60 min of global cardiac ischemia/reperfusion injury during CPB. Western blot analysis revealed that, after manipulation with different cardioplegic solutions, increased Bcl-2 expression and decreased cytochrome c expression were associated with cardiomyocytic apoptosis.

Conclusions: The NADH:ubiquinone oxidoreductase gene is up-regulated during global cardiac ischemia/reperfusion injury during CPB in canines. To our knowledge, involvement of this gene in global cardiac ischemia/reperfusion injury during CPB has not been described previously. The NADH:ubiquinone oxidoreductase gene may have a role in the regulation of molecular changes during the global cardiac ischemia/reperfusion injury during CPB, such as the up-regulation of Bcl-2, which might block release of cytochrome c from the mitochondria and prevent cardiomyocytic apoptosis.

MeSH terms

  • Animals
  • Blotting, Western
  • Cardiopulmonary Bypass
  • Cytochromes c / metabolism
  • Dogs
  • Electron Transport Complex I / genetics*
  • Electron Transport Complex I / metabolism
  • Gene Expression Regulation
  • Heart Arrest, Induced
  • Mitochondria, Heart / enzymology
  • Myocardial Reperfusion Injury / genetics*
  • Myocardial Reperfusion Injury / metabolism
  • Myocardium / enzymology*
  • Myocardium / metabolism
  • Polymerase Chain Reaction
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Up-Regulation

Substances

  • Proto-Oncogene Proteins c-bcl-2
  • Cytochromes c
  • Electron Transport Complex I