The production of hydroxyl radical by human neutrophils stimulated by arachidonic acid--measurements by ESR spectroscopy.
Human neutrophils incubated with sodium arachidonate generated hydroxyl radicals. The radical formed an adduct with the spin trap 5', 5-dimethyl-l-pyrroline-N-oxide (DMPO) and was subsequently detected by electron spin resonance (ESR) spectroscopy. The ESR signal was inhibited by mannitol and superoxide dismutase but not by catalase. Removal of glucose from the reaction mixture or the presence of glucose metabolic inhibitors including 2-deoxy-D-glucose and 3-O-methyl-D-glucose did not affect the production of hydroxyl radical by the neutrophils. The ESR signal was, however, inhibited by the lipoxygenase inhibitors nordihydroguaiaretic acid and N-ethylmaleimide. The involvement of lipoxygenase in the production of hydroxyl radical was demonstrated by the trapping of the radical with DMPO in a reaction mixture of soybean lipoxygenase and arachidonic acid (AA). These findings support our previous postulation that the metabolism of AA via the lipoxygenase pathway is a source of hydroxyl radical in stimulated neutrophils.
D019325:3-O-Methylglucose; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000494:pharmacology*; D002396:Catechols / Q000494:pharmacology; D003497:Cyclic N-Oxides; D003847:Deoxyglucose / Q000494:pharmacology; D004578:Electron Spin Resonance Spectroscopy; D005033:Ethylmaleimide / Q000494:pharmacology; D006801:Humans; D006878:Hydroxides / Q000097:blood*; D017665:Hydroxyl Radical; D016859:Lipoxygenase Inhibitors; D008353:Mannitol / Q000494:pharmacology; D009637:Masoprocol; D008757:Methylglucosides / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D013113:Spin Labels; D013482:Superoxide Dismutase / Q000494:pharmacology
D001095:Arachidonic Acids; D002396:Catechols; D003497:Cyclic N-Oxides; D006878:Hydroxides; D016859:Lipoxygenase Inhibitors; D008757:Methylglucosides; D013113:Spin Labels; D019325:3-O-Methylglucose; D016718:Arachidonic Acid; D017665:Hydroxyl Radical; D008353:Mannitol; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D009637:Masoprocol; D003847:Deoxyglucose; D013482:Superoxide Dismutase; D005033:Ethylmaleimide