Cellular mechanism of U78517F in the protection of porcine coronary artery endothelial cells from oxygen radical-induced damage.
1. The aim of this study was to clarify the role of lipid peroxidation in cellular injury as assessed by lactate dehydrogenase (LDH) release from cultured coronary artery endothelial cells of the pig. Cells exposed to H2O2 at concentrations of 0.1 to 20 mM or to a xanthine and xanthine oxidase (X/XO) reaction mixture released LDH into the medium. Significant release from X/XO-treated cells took place with a delay of 2 h. 2. Superoxide dismutase (SOD), catalase or dimethylthiourea attenuated the release of LDH from X/XO-treated cells. Similarly the putative inhibitor of lipid peroxidation, U78517F attenuated the release of LDH by X/XO with an IC50 of 0.08 microM. 3. H2O2 was continuously produced by the addition of X/XO to the medium alone. However, in the presence of endothelial cells, H2O2 was eliminated at 1 h. U78517F had no effect on either process. 4. The oxygen radical-induced release of LDH was associated with malondialdehyde (MDA) formation. U78517F inhibited the formation of MDA with an IC50 of 0.27 microM. 5. Reduction of the Ca2+ concentration in the incubation medium from 1.6 mM to 0.016 mM markedly attenuated the release of LDH from endothelial cells. Nifedipine (1 microM) did not attenuate the LDH release from the cells. 6. It is likely that porcine coronary artery endothelial cells can be thus injured by oxygen radicals presumably through hydroxyl radicals formed and consequent lipid peroxidation, and that the extracellular Ca2+ concentration plays an important role in the genesis of such endothelial cell damage.
Maeda
K
K
Department of Pharmacology, Upjohn Pharmaceuticals Limited, Tsukuba Research Laboratories, Ibaraki, Japan.
Kimura
M
M
Hayashi
S
S
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002839:Chromans / Q000494:pharmacology*; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D016166:Free Radical Scavengers; D005609:Free Radicals; D006861:Hydrogen Peroxide / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008315:Malondialdehyde / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects; D009543:Nifedipine / Q000494:pharmacology; D010084:Oxidation-Reduction; D010100:Oxygen / Q000633:toxicity*; D010879:Piperazines / Q000494:pharmacology*; D013552:Swine; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D014970:Xanthines / Q000494:pharmacology
D000975:Antioxidants; D002839:Chromans; D016166:Free Radical Scavengers; D005609:Free Radicals; D010879:Piperazines; D014970:Xanthines; C066382:U 78517F; D008315:Malondialdehyde; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D014969:Xanthine Oxidase; D009543:Nifedipine; D010100:Oxygen; D002118:Calcium