1,3-DIOCTANOYLGLYCEROL MODULATES ARACHIDONATE MOBILIZATION IN HUMAN NEUTROPHILS AND ITS INHIBITION BY PGBX - EVIDENCE OF A PROTEIN-KINASE-C-INDEPENDENT ROLE FOR DIACYLGLYCEROLS IN SIGNAL-TRANSDUCTION

Preincubation of human neutrophils with 1-oleoyl-2-acetylglycerol (OAG) enhances subsequent f-Met-Leu-Phe (fMLP)-stimulated arachidonate mobilization. We have recently demonstrated that preincubation of neutrophils with OAG also reverses inhibition of A23187 stimulated [H-3]arachidonate mobilization by the phospholipase A, inhibitors, PGBx and aristolochic acid. The present study has compared the effects of 1,2-sn-dioctanoylglycerol (1,2-diC8) and 1,3-dioctanoylglycerol (1,3-diC8) on these cellular events. Dose-dependent priming (ED50 < 2.5 muM) of fMLP-stimulated [H-3]arachidonate mobilization is obtained with both 1,2-diC8 and 1,3-diC8. Both diC8s also enhance fMLP-stimulated synthesis of leukotriene B4, 5-hydroxyeicosatetraenoic acid and platelet-activating factor, and generation of superoxide. Furthermore, both 1,2-diC8 and 1,3-diC8 reverse the effects of PGBx on A23187-stimulated [H-3]arachidonate mobilization and platelet-activating factor synthesis. By contrast, higher concentrations (5-10 muM) of 1,2-diC8, but not 1,3-diC8, directly stimulate both [H-3]arachidonate mobilization and superoxide generation. Since 1,3-diC8 does not activate protein kinase C (PKC), these results suggest that PKC is involved in direct activation of neutrophils by diacylglycerols but not in priming. Furthermore, reversal of the inhibitory effects of PGBx by diacylglycerols also appears to involve a PKC-independent mechanism.