Hypoxia/reoxygenation increases the permeability of endothelial cell monolayers: role of oxygen radicals.
We assessed the effect of hypoxia/reoxygenation on 14C-albumin flux across endothelial monolayers. Cultured bovine pulmonary artery endothelial cells were grown to confluence on nitrocellulose filters (pore size 12 microns). The endothelialized filters were mounted in Ussing-type chambers which were filled with cell culture medium (M 199). Equimolar amounts (33 nM) of 14C-labeled and unlabeled albumin were added to the "hot" and "cold" chambers, respectively. The monolayers were then exposed to successive periods (90 min) of normoxia (pO2 145 mmHg), hypoxia (pO2 20 mmHg), and reoxygenation (pO2 145 mmHg). A gas bubbling system was used to control media pO2 and to ensure adequate mixing. Four aliquots of culture media were taken during each period in order to calculate the 14C-albumin permeability across the endothelialized filter. In some experiments, either the xanthine oxidase inhibitor, oxypurinol (10 microM), or superoxide dismutase (600 U/mL), was added to the media immediately prior to the experiments. As compared to the normoxic control period, albumin permeability was 1.5 times higher during hypoxia (p less than 0.01) and 2.3 times higher during reoxygenation (p less than 0.01). The reoxygenation-induced increase in albumin permeability was prevented by either oxypurinol or superoxide dismutase. These data indicate that xanthine oxidase-derived oxygen radicals contribute to the hypoxia/reoxygenation-induced endothelial cell dysfunction. The altered endothelial barrier function induced by hypoxia/reoxygenation is consistent with the microvascular dysfunction observed following reperfusion of ischemic tissues.
Inauen
W
W
Department of Physiology, Louisiana State University Medical Center, Shreveport 71130.
Payne
D K
DK
Kvietys
P R
PR
Granger
D N
DN
D000818:Animals; D001692:Biological Transport; D002250:Carbon Radioisotopes; D002417:Cattle; D002478:Cells, Cultured; D003327:Coronary Disease / Q000378:metabolism*; D004727:Endothelium / Q000187:drug effects / Q000378:metabolism; D005609:Free Radicals; D010100:Oxygen / Q000378:metabolism*; D010117:Oxypurinol / Q000494:pharmacology*; D012709:Serum Albumin / Q000378:metabolism; D013482:Superoxide Dismutase / Q000494:pharmacology*; D014969:Xanthine Oxidase / Q000494:pharmacology*
D002250:Carbon Radioisotopes; D005609:Free Radicals; D012709:Serum Albumin; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D010117:Oxypurinol; D010100:Oxygen