Hydroperoxide-induced oxidative stress alters pyridine nucleotide metabolism in neonatal heart muscle cells.
An oxidant burden established by hydrogen peroxide (H2O2) overload may elicit postischemic myocardial damage. We assess herein the influence of H2O2-induced oxidative stress on heart muscle pyridine nucleotide metabolism. Exposure of neonatal rat cardiomyocytes to 50 microM-1.0 mM H2O2 bolus rapidly shifted their pyridine-nucleotide redox balance toward oxidation. At least 30% of the observed NADPH oxidation was independent of glutathione cycle activity and appeared chemical in nature with H2O2 itself, and not a radical metabolite, acting as oxidant. Cell exposure to H2O2 also depleted cardiomyocyte pyridine nucleotides as a consequence of enhanced utilization. The oxidative stress activated one major route of pyridine nucleotide catabolism (i.e., protein ADP-ribosylation) without acute inhibitory effect upon the other (cleavage by NAD glycohydrolase). The limited NAD sparing by metal chelators and inhibitors of ADP-ribosylation reflected pyridine nucleotide utilization for repair of single-strand DNA breaks caused by hydroxyl-like radicals formed intracellularly through iron-dependent H2O2 reduction. Cardiomyocyte NAD depletion during H2O2-induced oxidative stress was independent of cell integrity and lipid peroxidation. The NAD lost after a discrete H2O2 "pulse" was only partly replenished over a 24-h postinjury period. These data demonstrate that cardiomyocyte pyridine nucleotide metabolism is a nonperoxidative injury target that is chronically affected by H2O2 overload. Derangement of myocardial pyridine nucleotide pools due to oxidative stress may contribute to ischemic heart injury in vivo by interfering with cardiac hydrogen metabolism and redox balance.
Research Department, Ciba-Geigy Corporation, Summit, New Jersey 07901.
D R
DR
Janero
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D
Hreniuk
H M
HM
Sharif
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KC
Prout
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004249:DNA Damage; D005978:Glutathione / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D009243:NAD / Q000172:deficiency; D009244:NAD+ Nucleosidase / Q000378:metabolism; D009249:NADP / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D013312:Stress, Physiological / Q000139:chemically induced* / Q000378:metabolism*
D009243:NAD; D009249:NADP; D006861:Hydrogen Peroxide; D011065:Poly(ADP-ribose) Polymerases; D009244:NAD+ Nucleosidase; D005978:Glutathione