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Kinetics of nitric oxide and hydrogen peroxide production and formation of peroxynitrite during the respiratory burst of human neutrophils
Author(s) -
Carreras Maria Cecilia,
Pargament Griselda A.,
Catz Sergio D.,
Poderoso Juan J.,
Boveris Alberto
Publication year - 1994
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(94)80241-6
Subject(s) - peroxynitrite , hydrogen peroxide , chemistry , nitric oxide , superoxide , respiratory burst , chemiluminescence , superoxide dismutase , phorbol , omega n methylarginine , biochemistry , reactive oxygen species , oxygen , luminol , kinetics , liberation , neutrophile , nitric oxide synthase , antioxidant , enzyme , protein kinase c , chromatography , in vitro , organic chemistry , physics , quantum mechanics
Nitric oxide ( . NO) release, oxygen uptake and hydrogen peroxide (H 2 O 2 ) production elicited by increasing phorbol 12‐myristate 13‐acetate (PMA) concentrations were measured in human neutrophils. Half‐maximal activities were sequentially elicited at about 0.0001–0.001 μg ( . NO) and 0.001‐0.01 μg (H 2 O 2 ). At saturated PMA concentrations, . NO production, oxygen uptake and H 2 O 2 release were 0.56 ± 0.04, 3.32 ± 0.52 and 1.19±0.17 nmol · min −1 · 10 6 cells −1 . . NO production accounts for about 30% of the total oxygen uptake. Luminol‐dependent chemiluminescence, reported to detect NO reactions in other inflammatory cells, was also half‐maximally activated at about 0.001‐0.01 μg . Preincubation with N G ‐monomethyl‐ l ‐arginine ( l ‐NMMA) decreased O 2 uptake and . NO release but increased H 2 O 2 production, while superoxide dismutase (SOD) increased . NO detection by 30%. Chemiluminescence was also reduced by preincubation with l ‐NMMA and/or SOD. The results indicate that . NO release is part of the integrated response of stimulated human neutrophils and that, in these cells, kinetics of ″NO and O 2 .− release favour the formation of other oxidants like peroxynitrite.