Contribution of nitric oxide synthase to luminol-dependent chemiluminescence generated by phorbol-ester-activated Kupffer cells.
Phorbol 12-myristate 13-acetate-induced luminol chemiluminescence in rat Kupffer cells was doubled by the addition of L-arginine and significantly (up to 70%) inhibited by NG-nitro-L-arginine and NG-monomethyl-L-arginine, competitive inhibitors of L-arginine-dependent nitric oxide (NO) formation. The release of superoxide anion (O2-) by NADPH oxidase was neither affected by L-arginine nor by the inhibitors. Only very slight luminol chemiluminescence was detectable in lipopolysaccharide-pretreated Kupffer cells, a condition in which significant amounts of NO were formed but no O2-. In a cell-free system, significant luminol chemiluminescence only occurred when both authentic NO and the O2-/H2O2- generating system xanthine/xanthine oxidase were present. The results indicate that luminol chemiluminescence in phorbol-ester-activated Kupffer cells largely depends on L-arginine metabolism by NO synthase, requiring the concurrent formation of NO and O2-/H2O2.
Wang
J F
JF
Institut für Physiologische Chemie I, Heinrich-Heine-Universität Düsseldorf, Federal Republic of Germany.
Komarov
P
P
Sies
H
H
de Groot
H
H
D000166:Acridines / Q000378:metabolism; D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D002374:Catalase / Q000378:metabolism; D002474:Cell-Free System; D066298:In Vitro Techniques; D007728:Kupffer Cells / Q000187:drug effects* / Q000378:metabolism; D008163:Luminescent Measurements; D008165:Luminol / Q000378:metabolism*; D008297:Male; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D019255:NADPH Oxidases; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase; D019335:Nitroarginine; D051381:Rats; D011919:Rats, Inbred Strains; D013482:Superoxide Dismutase / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D019820:Xanthine; D014969:Xanthine Oxidase / Q000378:metabolism; D014970:Xanthines / Q000378:metabolism; D019323:omega-N-Methylarginine
D000166:Acridines; D014970:Xanthines; D019820:Xanthine; D019335:Nitroarginine; C033472:10,10'-dimethyl-9,9'-biacridinium; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D008165:Luminol; D001120:Arginine; D002374:Catalase; D019001:Nitric Oxide Synthase; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D000594:Amino Acid Oxidoreductases; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D013755:Tetradecanoylphorbol Acetate