Oxidant generation with K(+)-induced depolarization in the isolated perfused lung.
This study evaluated whether cell membrane depolarization can induce oxidant generation in the isolated perfused rat lung as has been demonstrated with bovine pulmonary artery endothelial cells. Depolarization was produced by perfusing the lungs with high [K+] or with glyburide and was evaluated with bis-oxonol lung surface fluorometry. Lung surface bis-oxonol fluorescence increased above baseline (at 5.9 mM K+) by 18.5% with 24 mM K+, 35% with 48 mM K+, and 67% with 96 mM K+, indicating graded membrane depolarization, and by 75% during perfusion with 10 microM glyburide. Oxidant generation was evaluated with hydroethidine lung surface fluorometry, and with assay of tissue thiobarbituric acid reactive substance (TBARS), conjugated dienes, and perfusate H2O2. Depolarization by high K+ or glyburide led to significant increases in generation of tissue oxidants and lipid peroxidation. Bodipy-FL-glyburide microfluorography showed localization of glyburide binding primarily to vascular endothelial cells vascular and airway smooth muscle cells, alveolar type II cells, and to nonciliated cells of the airway epithelium. These results indicate that cellular depolarization is associated with oxidant generation by the lung and suggests a role for K(+)-channels in these events.
al-Mehdi
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Institute for Environmental Medicine, University of Pennsylvania Medical Center, Philadelphia 19104, USA.
Shuman
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Fisher
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D000818:Animals; D005456:Fluorescent Dyes / Q000378:metabolism; D005470:Fluorometry; D005905:Glyburide / Q000378:metabolism / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000378:metabolism; D066298:In Vitro Techniques; D015227:Lipid Peroxidation; D008168:Lung / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D018613:Microscopy, Confocal; D016877:Oxidants / Q000378:metabolism*; D010477:Perfusion; D010617:Phenanthridines / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology*; D015221:Potassium Channels / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013858:Thiobarbiturates / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis
D005456:Fluorescent Dyes; D016877:Oxidants; D010617:Phenanthridines; D015221:Potassium Channels; D013858:Thiobarbiturates; D017392:Thiobarbituric Acid Reactive Substances; C058428:hydroethidine; C025284:bis(1,3-diethylthiobarbiturate)trimethineoxonol; D006861:Hydrogen Peroxide; D011188:Potassium; D005905:Glyburide