Xanthine oxidase-induced injury to endothelium: role of intracellular iron and hydroxyl radical.
The major objective of the present study was to characterize the sequence of events leading to endothelial cytotoxicity induced by oxidants generated extracellularly by xanthine oxidase. 51Cr-labeled monolayers of calf pulmonary artery endothelial cells were exposed to a reaction mixture containing hypoxanthine, xanthine oxidase, and chelated iron (HX/XO) and endothelial cell injury was quantitated as 51Cr release into the media. Catalase, but not mannitol or superoxide dismutase, prevented endothelial cell injury induced by HX/XO, indicating that H2O2 was the mediator of the cytotoxicity. Pretreatment of the cells with free deferoxamine (an iron chelator), but not with deferoxamine bound to dextran (mol wt 40,000), prevented endothelial cell injury induced by HX/XO or H2O2. Of the membrane-permeant hydroxyl radical scavengers dimethylsulfoxide and dimethylthiourea, only dimethylthiourea prevented 1) HX/XO or H2O2-induced endothelial cytotoxicity and 2) deoxyribose degradation by hydroxyl radicals (.OH) generated by an iron-catalyzed reaction on the sugar (site-specific reaction). The concentration of ferritin required to produce significant quantities of .OH was much greater than that present in endothelial cells, and ferritin-catalyzed .OH formation was not affected by deferoxamine, indicating that ferritin-bound iron is most likely not the physiologically active catalyst. We conclude that extracellularly generated H2O2 can enter the cell and interact with nonferritin iron to produce the cytotoxic .OH via a site-specific reaction.
Kvietys
P R
PR
Department of Physiology, Louisiana State University Medical Center, Shreveport 71130-3932.
Inauen
W
W
Bacon
B R
BR
Grisham
M B
MB
D000818:Animals; D003676:Deferoxamine / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000473:pathology; D005609:Free Radicals; D006861:Hydrogen Peroxide / Q000494:pharmacology; D006878:Hydroxides / Q000378:metabolism*; D017665:Hydroxyl Radical; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000494:pharmacology; D007425:Intracellular Membranes / Q000378:metabolism*; D007501:Iron / Q000502:physiology*; D008315:Malondialdehyde / Q000378:metabolism; D014969:Xanthine Oxidase / Q000494:pharmacology*
D005609:Free Radicals; D006878:Hydroxides; D007042:Hypoxanthines; D019271:Hypoxanthine; D017665:Hydroxyl Radical; D008315:Malondialdehyde; D006861:Hydrogen Peroxide; D007501:Iron; D014969:Xanthine Oxidase; D003676:Deferoxamine