Effect of extracellular calcium on superoxide release by rat alveolar macrophages

Alveolar macrophages can be stimulated to release superoxide into the extracellular medium. The mechanism of activation of superoxide release has been reported to be mediated by the movement of Ca2+ into the cytosol, but the involvement of extracellular Ca2+ in this process has remained uncertain. Extracellular Ca2+ was not an absolute requirement for activation of superoxide release; however, 1.3 mM extracellular Ca2+ caused an approximate twofold enhancement of superoxide release stimulated by either concanavalin A or formyl-methionyl-leucyl-phenylalanine but had no effect on digitonin-stimulated superoxide release. Concanavalin A-stimulated superoxide was inhibited by the "Ca2+ entry blockers," Mn2+ (1 microM) or methoxyverapamil (D-600) (100 microM). Inhibition by Mn2+ was competitive with extracellular Ca2+, whereas inhibition by D-600 was noncompetitive. Neither inhibitor, however, affected superoxide release after initiation. Thus activation of superoxide release by alveolar macrophages can be accomplished, in part, through the entrance of Ca2+ into the cell.