Nitric oxide does not contribute to superoxide-mediated sulfhydryl oxidation in human polymorphonuclear leukocytes.
Recent findings have suggested that nitric oxide (NO) reacts with superoxide anion (O2-) to form a potential oxidant, peroxynitrite anion, which then decays to hydroxyl radical and nitrogen dioxide. In order to ascertain this hypothesis in human polymorphonuclear leukocytes (PMNs) which release both NO and O2-, we studied oxidation of L-cysteine (CYS) and bovine serum albumin (BSA) by PMNs and cell-free O2(-)-generating system of hypoxanthine (HX)-xanthine oxidase (XO) reaction. Oxidation of CYS by HX-XO was equally inhibited by superoxide dismutase (SOD) and catalase (CAT), and that of BSA by HX-XO was inhibited weakly by SOD and strongly by CAT. PMNs stimulated with phorbol 12-myristate 13-acetate increased the oxidation rates of CYS and BSA, and they were inhibited by SOD and CAT almost in a similar way to those by HX-XO. The NO synthase inhibitor, NG-monomethyl-L-arginine (NMMA), was confirmed to have an inhibitory effect on the inhibition of platelet aggregation by PMNs, and L-arginine (ARG) reversed this effect. However, pretreatment of PMNs with either of NMMA, or ARG, or both did not change the oxidation rates of CYS and BSA. We could not confirm the hypothesis at least in human PMNs that interaction of NO with O2- forms powerful oxidants to sulfhydryls of CYS and BSA. These results suggest that oxidation of sulfhydryls of CYS and BSA by PMNs is primarily dependent on reactive oxygen species, and is not modified by NO production.
Koshida
H
H
National Biomedical Center for Spin Trapping and Free Radicals, Oklahoma Medical Research Foundation, Oklahoma City.
Kotake
Y
Y
D000328:Adult; D006801:Humans; D008297:Male; D008875:Middle Aged; D009504:Neutrophils / Q000378:metabolism*; D009569:Nitric Oxide / Q000502:physiology*; D010084:Oxidation-Reduction; D013438:Sulfhydryl Compounds / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D013438:Sulfhydryl Compounds; D013481:Superoxides; D009569:Nitric Oxide; D013755:Tetradecanoylphorbol Acetate