Conditions to study nitric oxide generation by polymorphonuclear cells from an inflammatory exudate in rats.
Superoxide and nitric oxide release by leukocytes has been usually performed after exposure to a particular stimulus. We measured the generation of superoxide and nitric oxide by cells isolated from an inflammatory exudate of rats in either the absence or the presence of a variety of stimuli. Nonstimulated leukocytes generated superoxide radical (1.2 nmol x 10(6) cells(-1)) and nitric oxide (3.8 nmol x 10(6) cells(-1)) after 2 h incubation. When cells were incubated with lipopolysaccharides, opsonized zymosan or phorbol 12-myristate 13-acetate, superoxide level increased while nitric oxide decreased. Phorbol 12-myristate 13-acetate (100 ng/ml) induced a decrease of 0.88 nmol x 10(6) cells(-1) compared with nonstimulated cells, and incubation with N-iminoethyl-L-ornithine increased superoxide production by 0.81 nmol x 10(6) cells(-1). These results provide clear evidence that cells from an inflammatory exudate which are already triggered are able to generate a considerable amount of nitric oxide and in less proportion superoxide, that the measure of nitric oxide must be performed without a further stimulus, and that both molecules react in an equimolar proportions to give peroxynitrite anion.
Ródenas
J
J
Departament de Fisiologia, Facultat de Biologia, Universitat de Barcelona, Spain.
Carbonell
T
T
Mitjavila
M T
MT
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002351:Carrageenan; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005122:Exudates and Transudates; D007249:Inflammation / Q000503:physiopathology*; D007700:Kinetics; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D019335:Nitroarginine; D009952:Ornithine / Q000031:analogs & derivatives / Q000494:pharmacology; D051381:Rats; D013481:Superoxides / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D015054:Zymosan / Q000494:pharmacology
D004791:Enzyme Inhibitors; D013481:Superoxides; D019335:Nitroarginine; D009569:Nitric Oxide; C065027:N(G)-iminoethylornithine; D002351:Carrageenan; D015054:Zymosan; D001120:Arginine; D009952:Ornithine; D019001:Nitric Oxide Synthase; D013755:Tetradecanoylphorbol Acetate