Luminol chemiluminescence in rat macrophages and granulocytes: the role of NO, O2-/H2O2, and HOCl.
Luminol chemiluminescence was increased up to five-fold by L-arginine and markedly inhibited by NG-nitro-L-arginine (L-NNA) in phorbol ester (PMA) or opsonized zymosan-activated rat Kupffer cells, and in PMA-activated rat peritoneal and alveolar macrophages. While in Kupffer cells these effects did occur without pretreatment with lipopolysaccharides (LPS), LPS pretreatment was a requirement in peritoneal and alveolar macrophages. Azide (0.05 mM) had no effect on luminol chemiluminescence in the macrophages. The changes in luminol chemiluminescence were accompanied by parallel changes in nitric oxide (NO) formation. Macrophage superoxide anion radical (O2-) production was not significantly changed by addition of L-arginine and L-NNA nor by pretreatment with LPS. No hypochlorous acid (HOCl) formation was detectable in the macrophages. In contrast, in rat granulocytes activated by a variety of stimuli including PMA, zymosan, the chemotactic peptide formyl-methionyl-leucyl-phenylalanine and the calcium ionophore A23187 with or without pretreatment with LPS, L-arginine and L-NNA had no effect on luminol chemiluminescence. Luminol chemiluminescence, however, was largely inhibited by 0.05 mM azide. The activated granulocytes released significant amounts of HOCl but did not generate NO. These results demonstrate that NO may largely contribute to luminol chemiluminescence in rat macrophages, in which HOCl formation does not occur. On the other hand, HOCl is the reactive oxygen species responsible for luminol chemiluminescence in rat granulocytes, where NO is formed only in minor quantities, if at all.
Klinische Forschergruppe Leberschädigung, Heinrich-Heine-Universität, Düsseldorf, Federal Republic of Germany.
J F
JF
Wang
P
P
Komarov
H
H
de Groot
D000818:Animals; D001120:Arginine / Q000378:metabolism; D001386:Azides / Q000494:pharmacology; D000001:Calcimycin / Q000494:pharmacology; D002374:Catalase / Q000378:metabolism; D006098:Granulocytes / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D006997:Hypochlorous Acid / Q000378:metabolism*; D007728:Kupffer Cells / Q000378:metabolism; D008163:Luminescent Measurements; D008165:Luminol; D008264:Macrophages / Q000378:metabolism*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D051381:Rats; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D001386:Azides; D013481:Superoxides; D009569:Nitric Oxide; D000001:Calcimycin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D008165:Luminol; D006997:Hypochlorous Acid; D001120:Arginine; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D013755:Tetradecanoylphorbol Acetate