Hydrogen peroxide stimulates both prostanoid and platelet-activating factor (PAF) biosynthesis in cultured rat mesangial cells and isolated rat glomeruli. The present experiments were designed to try to establish some relationship between prostanoid and PAF synthesis in these renal structures, in the presence of hydrogen peroxide. Cells and glomeruli were incubated with hydrogen peroxide under different experimental conditions, and thromboxane B2 (TXB2), the stable metabolite of thromboxane A2 (TXA2), and prostaglandin E2 (PGE2) concentrations were measured in the supernatants of the cells or glomeruli. Moreover, H2O2-dependent PAF synthesis was measured by high performance liquid chromatography (HPLC) ([3H]acetate incorporation) and radioimmunoassay. H2O2 induced increased TXB2 and PGE2 production in cultured rat mesangial cells and isolated rat glomeruli. This effect was blocked by incubation in the presence of a PAF-receptor antagonist, BN-52021. This antagonist has no intrinsic effect either in basal prostanoid synthesis or in arachidonic acid-stimulated glomerular TXB2 synthesis. Alprazolam, another PAF antagonist, nonchemically related to BN-52021, also completely blocked the H2O2-induced production of TXB2 by isolated rat glomeruli. Moreover, H2O2 was also able to induce an increased [3H]acetate incorporation into a fraction comigrating with a PAF standard in HPLC in isolated glomeruli, and this effect was dependent on the H2O2 concentration tested. Moreover, H2O2 was also able to induce an increased [3H]acetate incorporation and increased synthesis of radioimmunoassayable PAF in cultured mesangial cells. These results suggest that the increased synthesis of PGE2 and TXB2 induced by H2O2 could be dependent on platelet-activating factor production.