prostaglandin-d2 and 4-bromophenacyl-bromide

The contribution of phospholipases A2 (PLA2) and D (PLD) activation to arachidonic acid liberation and prostaglandin D2 (PGD2) formation was studied in stimulated rat peritoneal mast cells. Stimulation of the cells with ionomycin induced time-dependent and Ca(2+)-concentration-dependent increase in arachidonic acid liberation and PGD2 formation, and the Ca(2+)-dependent increase was especially remarkable at extracellular Ca2+ concentration higher than 200 microM. Staurosporine did not induce any effect on the arachidonic acid liberation, indicating that protein kinase C is not involved in the liberation. Addition of ethanol to the cells decreased the ionomycin-stimulated arachidonic acid liberation to 40% of the control, while it decreased the PGD2 formation almost completely, with the increase in phosphatidylethanol formation. Propranolol, a phosphatidate phosphohydrolase inhibitor, caused similar effects. p-Bromophenacyl bromide, a PLA2 inhibitor, inhibited partially the arachidonic acid liberation. The inhibition of the liberation by combination of p-bromophenacyl bromide and ethanol was additive and reached approximately 90%. Under the conditions used p-bromophenacyl bromide did not influence significantly the PLD activity assessed by the phosphatidylethanol formation. Histamine release was decreased by ethanol treatment to 35% of the control. These results suggest that more than half of the total arachidonic acid liberation is mediated by the sequential pathway of PLD/phosphatidate phosphohydrolase/diacylglycerol lipase and more than half of histamine release is also dependent on PLD activation, while the PGD2 formation is fully mediated by the pathway. PLA2 also contributes to arachidonic acid liberation but to a lower extent.

Topics: Acetophenones; Animals; Arachidonic Acid; Calcium; Calcium Chloride; Cytosol; Ethanol; Histamine Release; In Vitro Techniques; Ionomycin; Kinetics; Mast Cells; Phospholipase D; Phospholipases A; Phospholipases A2; Prostaglandin D2; Rats; Rats, Wistar; Tritium

In isolated perfused rat liver, adenosine infusion (50 microM) led to increases in glucose output and portal pressure and a net K+ release of 3.7 +/- 0.21 mumol/g, which was followed by an equivalent net K+ uptake after cessation of the nucleoside infusion. These effects were accompanied by a transient stimulation of hepatic prostaglandin D2 and thromboxane B2 release. The Ca2+ release observed upon adenosine infusion (50 microM) was 23.5 +/- 5.2 nmol/g, i.e. 10-20% of the Ca2+ release observed with extracellular ATP (50 microM). Indomethacin (10 microM) prevented the adenosine-induced stimulation of glucose output and the increase in portal pressure by 79 and 63% respectively, and completely abolished the stimulation of prostaglandin D2 release. The thromboxane A2 receptor antagonist BM 13.177 (20 microM), the phospholipase A2 inhibitor 4-bromophenacyl bromide (20 microM) and the cyclo-oxygenase inhibitor ibuprofen (50 microM) also decreased the glycogenolytic and vasoconstrictive responses of the perfused rat liver upon adenosine infusion by 50-80%. When the indomethacin inhibition of adenosine-induced prostaglandin D2 release was titrated, a close correlation between prostaglandin D2 release and the metabolic and vascular responses to adenosine was observed. These findings suggest an important role for eicosanoids in mediating the nucleoside responses in the perfused rat liver. Since eicosanoids are known to be formed by non-parenchymal cells in rat liver [Decker (1985) Semin. Liver Dis. 5, 175-190], the present study gives further evidence for an important role of eicosanoids as signal molecules between the different liver cell populations.

Topics: Acetophenones; Adenosine; Animals; Blood Pressure; Glucose; Ibuprofen; Indomethacin; Liver; Male; Portal System; Prostaglandin D2; Rats; Rats, Inbred Strains; Secretory Rate; Sulfonamides; Thromboxane B2

4-Bromophenacyl bromide at a concentration of 50 microM does not inhibit phospholipase A2 activity in liver macrophages. Rather, this compound increases the amount of radioactivity released from [3H]arachidonate-prelabeled Kupffer cells and leads to the formation of small amounts of thromboxane, prostaglandin D2 and prostaglandin E2. Also the zymosan-induced formation of thromboxane and prostaglandin E2 from endogenous sources which is thought to involve phospholipase A2 remains unaffected in the presence of this compound. The generation of superoxide and the formation of prostaglandin D2 from arachidonate and after stimulation of the cells with zymosan, however, are blocked by 4-bromophenacyl bromide. Furthermore, this compound suppresses the incorporation of externally added arachidonate into membrane lipids of the cells. 4-Bromophenacyl bromide seems, therefore, not to be a useful tool to demonstrate the involvement of phospholipase A2 in complex biological systems.

Topics: Acetophenones; Animals; Arachidonic Acids; Cells, Cultured; Chromatography, High Pressure Liquid; Kupffer Cells; Membrane Lipids; Phospholipases; Phospholipases A; Phospholipases A2; Phospholipids; Prostaglandin D2; Prostaglandins E; Radioimmunoassay; Rats; Thromboxane B2; Zymosan