Cell alkalinization is not necessary and increased sodium influx is not sufficient for stimulated superoxide production

AbstractPreincubation of rabbit neutrophils for 5 min with the protein kinase C inhibitor H7 causes inhibition of the rise in intracellular pH but not the increase in Na+ influx or stimulated oxidative burst produced by the chemotactic factor formyl-methionyl-leucyl-phenylalanine. On the other hand, the stimulated superoxide production, but not the increase in Na+ influx produced by phorbol 12-myristate 13-acetate, is inhibited by H7. The effect is more pronounced on the rate than the extent of the stimulated superoxide release. Furthermore, cell acidification produced by the phorbol ester but not by the chemotactic factor is decreased in the presence of H7. These results suggest that (i) most of the stimulated Na+ influx is not coupled to H+ efflux, (ii) in the case of the chemoattractant, the rise in intracellular pH is not necessary for stimulated superoxide production, (iii) the increase in Na+influx, in the case of the phorbol ester, is not sufficient for the stimulation of the oxidative burst, and (iv) the sources of the H+ responsible for the stimulated pH drop are the various metabolic activities of the cell, including NADPH oxidation and activation of the hexose monophosphate shunt