Effect of hypoxia and reoxygenation on the formation and release of reactive oxygen species by porcine pulmonary artery endothelial cells.
Endothelial cells are critical targets in both hypoxia- and reoxygenation-mediated lung injury. Reactive O2 species (ROS) have been implicated in the pathogenesis of hypoxic and reoxygenation lung injury, and xanthine dehydrogenase/oxidase (XDH/XO) is a major generator of the ROS. Porcine pulmonary artery endothelial cells (PAEC) have no detectable XDH/XO. This study was undertaken to examine 1) ROS production by hypoxic porcine PAEC and their mitochondria and 2) ROS production and injury in reoxygenated PAEC lacking XDH/XO activity. Intracellular H2O2 generation and extracellular H2O2 and O2 divided release were measured after exposure to normoxia (room air-5% CO2), hypoxia (0% O2-95% N-5% CO2), or hypoxia followed by normoxia or hyperoxia (95% O2-5% CO2). Exposure to hypoxia results in significant reductions in intracellular H2O2 formation and extracellular release of H2O2 and O2 by PAEC and mitochondria. The reductions occur with as little as a 2 h exposure and progress with continued exposure. During reoxygenation, cytotoxicity was not observed, and the production of ROS by PAEC and their mitochondria never exceeded levels observed in normoxic cells. The absence of XDH/XO may prevent porcine PAEC from developing injury and increased ROS production during reoxygenation.
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Department of Medicine, University of Florida, Gainesville, USA.
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D000818:Animals; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000473:pathology; D005110:Extracellular Space / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D000860:Hypoxia / Q000378:metabolism* / Q000473:pathology; D008928:Mitochondria / Q000378:metabolism; D010100:Oxygen / Q000494:pharmacology*; D011651:Pulmonary Artery / Q000378:metabolism* / Q000473:pathology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013552:Swine
D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D010100:Oxygen