Superoxide Release by Zymosan‐Stimulated Rat Kupffer Cells in vitro

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Abstract
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 monophophate shunt were observed. Chemiluminescence and O2 production were almost completely inhibited by superoxide dismutase and iodoacetate. Zymosan‐induced chmiluminesceance 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.

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