Endothelial cell oxidant production: effect of NADPH oxidase inhibitors.
The effects of known leukocyte NADPH oxidase inhibitors on general cellular oxidant production in cultured human endothelial cells (EC) has been investigated. EC were stimulated with 10 nM phorbol 12-myristate 13-acetate and cellular oxidant production measured in the presence and absence of inhibitors that act on various substituents of the oxidase complex and its activation pathways. The effects of the cytosolic oxidase subunit translocation inhibitors, catechols (3,4-dihydroxybenzaldehyde, caffeic acid, and protocatechuic acid), ortho-methoxy-substituted catechols (apocynin, vanillin, and 4-nitroguaiacol), and quinone, 1,4-naphthoquinone; flavoprotein inhibitors, diphenylene iodonium and quinacrine; haem ligands, imidazole and pyridine; directly acting thiol reagents, disulfiram and penicillamine; NADPH analogue, Cibacron Blue; redox active inhibitors, quercetin and esculetin; intracellular calcium antagonist, TMB-8; and calmodulin antagonists, W-7 and trifluoperazine, were determined. All compounds reduced oxidant production in stimulated EC. These findings add to previous observations suggesting the presence of a functionally active NADPH oxidase in EC. Identifying the major cellular reactive oxygen species source in perturbed EC will provide new insights into our understanding of endothelial dysfunction, which has been hypothesized to be a major contributing factor in the pathogenesis of atherosclerosis.
Holland
J A
JA
Department of Biology, State University of New York College at Geneseo, 14454, USA. james.holland3@med.va.gov
O'Donnell
R W
RW
Chang
M M
MM
Johnson
D K
DK
Ziegler
L M
LM
D000975:Antioxidants / Q000494:pharmacology*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005420:Flavoproteins / Q000037:antagonists & inhibitors; D005456:Fluorescent Dyes; D006418:Heme / Q000037:antagonists & inhibitors; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors*; D016877:Oxidants / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014471:Umbilical Veins / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism
D000975:Antioxidants; D002121:Calcium Channel Blockers; D004791:Enzyme Inhibitors; D005420:Flavoproteins; D005456:Fluorescent Dyes; D016877:Oxidants; D017382:Reactive Oxygen Species; D013439:Sulfhydryl Reagents; D006418:Heme; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D013755:Tetradecanoylphorbol Acetate