Superoxide release by zymosan-stimulated rat Kupffer cells in vitro.
Kupffer cells were isolated from pronase-perfused rat livers and were maintained as a monolayer culture in a state of high purity and viability. Immediately after contact with zymosan particles, O2 uptake of the Kupffer cells increased fivefold; about 50% of the net oxygen consumed was accounted for as superoxide released into the medium. Concomitantly, a transient burst of luminol-dependent chemiluminescence, an increased activity of NAD(P)H oxidase and a stimulation of the flow of glucose through the hexose monophosphate shunt were observed. Chemiluminescence and O2- production were almost completely inhibited by superoxide dismutase and iodoacetate. Zymosan-induced chemiluminescence was not inhibited in the presence of the non-penetrating thiol reagents, 5,5'-dithio-bis-2-nitrobenzoate and iodoacetyl-sepharose. Iodoacetate acted on the cytosolic glucose-6-phosphate dehydrogenase rather than on NAD(P)H oxidase of the cell membrane.
D000818:Animals; D002478:Cells, Cultured; D004789:Enzyme Activation / Q000187:drug effects; D005260:Female; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D007728:Kupffer Cells / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008163:Luminescent Measurements; D009097:Multienzyme Complexes / Q000096:biosynthesis; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence; D013481:Superoxides / Q000378:metabolism*; D015054:Zymosan / Q000494:pharmacology*
D007461:Iodoacetates; D009097:Multienzyme Complexes; D013481:Superoxides; D015054:Zymosan; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D010100:Oxygen; D019807:Iodoacetic Acid