Platelet-activating factor stimulates eicosanoid production in cultured feline tracheal epithelial cells

The effect of platelet-activating factor (PAF) on eicosanoid generation and release in cultured feline tracheal epithelial cells was investigated by measuring a wide range of lipoxygenase and cyclooxygenase pathway products. Subconfluent epithelial cell cultures were stimulated by PAF and eicosanoid production was determined by high performance liquid chromatography (HPLC) of ³H]-labeled arachidonic acid (AA) metabolites and by radioimmunoassay (RIA) following HPLC separation. The HPLC chromatograms revealed that PAF augmented the release of prostaglandin (PG)E₂, PGF_2α, 12-hydroxyeicosatetraenoic acid (HETE), and AA. Among these eicosanoids, PGE₂ predominated under baseline conditions and following PAF exposure. RIAs of the nonradiolabeled HPLC elution corresponding to various eicosanoid standards demonstrated that PAF increased the production of 6-keto-PGF_1α, thromboxane B₂ (TXB₂), PGD₂, 5-HETE, and 15-HETE, as well as PGE₂, PGF_2α, and 12-HETE. The PAF-induced eicosanoid augmentation was dose-dependent and occurred within 1 hour with a prompt decline following termination of PAF exposure. This stimulating effect of PAF on eicosanoid release was blocked by two PAF receptor antagonists, Ro 19-3704 and WEB 2086. The PAF-induced increase in eicosanoid release was similar in magnitude to the increase caused by calcium ionophore (Ca-ionophore) A23187, a potent known stimulus for eicosanoid release. Cells of different culture durations (3 and 6 days) showed similar capacity for eicosanoid production. We conclude that PAF stimulates the production of cyclooxygenase and lipoxygenase pathway products from airway epithelial cells via PAF receptors, and that these epithelium-derived eicosanoids may be responsible for some of the PAF-induced respiratory physiological and pathophysiological effects.