Differential-display polymerase chain reaction identifies nicotinamide adenine dinucleotide-ubiquinone oxidoreductase as an ischemia/reperfusion-regulated gene in cardiomyocytes.
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.
Yeh
Chi-Hsiao
CH
Division of Thoracic and Cardiovascular Surgery, Chang Gung Memorial Hospital,Taiwan.
Pang
Jong-Hwei S
JH
Wu
Yi-Cheng
YC
Wang
Yao-Chang
YC
Chu
Jaw-Ji
JJ
Lin
Pyng Jing
PJ
D000818:Animals; D015153:Blotting, Western; D002315:Cardiopulmonary Bypass; D045304:Cytochromes c / Q000378:metabolism; D004285:Dogs; D042967:Electron Transport Complex I / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006324:Heart Arrest, Induced; D008929:Mitochondria, Heart / Q000201:enzymology; D015428:Myocardial Reperfusion Injury / Q000235:genetics* / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
D019253:Proto-Oncogene Proteins c-bcl-2; D045304:Cytochromes c; D042967:Electron Transport Complex I