Phorbor Myristate Acetate Induces NADPH Oxidase Activity of Cytochalasin B-Primed Neutrophils through the Protein Kinase C-Independent Pathway

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We examined the effect of cytochalasin B (CB) or granulocyte colony stimulating factor (GCSF) on superoxide radical (O_2^-) production of neutrophils by phorbor myristate acetate (PMA)-stimulation. It was observed that O_2^- generation of intact and GCSF-treated neutrophils by PMA-stimulation showed a lag during the early stage, and was largely inhibited by 1-(5-isoquinoline-sulfonyl)-3-methyl-piperazine (200 μM) or GF109203X (GFX) (0.2 μM), but not by ethanol (1%) and wortmannin (100 nM). In contrast, O_2^- generation of CB-pretreated neutrophils by PMA-stimulation did not show a lag, but was less than that of intact cells, and was only minimally depressed by the above inhibitors, but was markedly depressed by the simultaneous addition of GFX and ethanol or GFX and wortmannin. Although translocation of p47phox and p67phox to the membrane fraction by PMA-stimulation of intact and GCSF-treated neutrophils occurred in parallel with O_2^- production, that of CB-treated neutrophils by PMA-stimulation was not always proportional to O_2^- production. These findings suggest that pretreatment of neutrophils with CB dramatically alters the PMA response of the cells; that is, the protein kinase C-dependent pathway is largely depressed, and a phospholipase D-dependent one for NADPH oxidase activation appears in CB-treated cells.
公益社団法人日本薬学会の論文
2000-09-01