Effect of hyperoxia and exogenous oxidant stress on pulmonary artery endothelial cell Na+/H+ antiport activity.
Little is known about the mechanisms of altered cell membrane function after hyperoxic exposure. We determined the effects of hyperoxic exposure and exogenous oxidant stress with xanthine/xanthine oxidase (X/XO) on Na+/H+ antiport activity. Pulmonary artery endothelial cell monolayers were incubated in 95% O2/5% CO2 (24 to 72 hours) simultaneously with controls placed in 21 % O2/5% CO2. Monolayers were then incubated for 2 hours in MEM with or without X/XO (100 micromol/L X; 0.01 U/ml XO). Antiport activity was determined as the rate of recovery from intracellular acidosis by measurement of intracellular pH (pH,) with 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF). Hyperoxic exposure (72 hours) decreased Na+/H+ antiport activity as compared with that in control monolayers. Exogenous oxidant stress also decreased antiport activity in both control and hyperoxic cells, but this effect was more pronounced in hyperoxic cells at all time points. These changes occurred in the absence of overt cytotoxicity. Incubation with antioxidants (polyethylene glycol-superoxide dismutase (PEG-SOD), PEG-catalase, vitamin E), N-acetylcysteine, or phospholipase A2 (PLA2) inhibitors did not prevent the decrease in antiport activity after hyperoxic exposure. Conditioned medium experiments demonstrated that the diminished antiport activity was not related to release of a soluble mediator after hyperoxic exposure. These findings suggest that the diminished Na+/H+ antiport activity represents a sublethal form of membrane dysfunction that may be a component of the increased endothelial cell susceptibility to injury after hyperoxic exposure.
Cutaia
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Department of Medicine, VA Medical Center, Brown University School of Medicine, Providence, RI 02908, USA.
Parks
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D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002857:Chromium / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000633:toxicity; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D018384:Oxidative Stress / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000502:physiology*; D017923:Sodium-Hydrogen Exchangers / Q000187:drug effects / Q000502:physiology*; D019820:Xanthine; D014969:Xanthine Oxidase / Q000494:pharmacology; D014970:Xanthines / Q000494:pharmacology
D017077:Culture Media, Conditioned; D017923:Sodium-Hydrogen Exchangers; D014970:Xanthines; D002857:Chromium; D019820:Xanthine; D014969:Xanthine Oxidase; D010100:Oxygen