Production of platelet-activating factor by human vascular endothelial cells: evidence for a requirement for specific agonists and modulation by prostacyclin.

Abstract

Primary cultures of confluent endothelial cells derived from human umbilical veins produce platelet-activating factor (PAF) (1-alkyl-2-acetyl-sn-glycero-3-phosphocholine) when stimulated with appropriate agonists. Highly purified human thrombin and calcium ionophore A23187 stimulate the incorporation of [3H] acetate into a lipid product that has been identified as PAF by its behavior in thin-layer chromatographic and high-performance liquid chromatographic systems, the presence of characteristic biologic activity, and appropriate response to phospholipases. A number of other humoral mediators, examined because they directly influence the activity of vascular cells or because they may mediate endothelial injury, do not stimulate PAF production by endothelial cells. This indicates that the synthesis of PAF by cultured human endothelial cells is a response to specific agonists and is not an unregulated event that occurs as a result of nonspecific cellular perturbation. The PAF produced by thrombin-treated endothelial cells is a potent stimulus for platelet activation, as assayed by the aggregation of human platelets in autologous plasma. The production of PAF by endothelial monolayers is attenuated by prostacyclin, another product of stimulated endothelial cells. Conversely, PAF production is enhanced by treatment of the endothelial cells with indomethacin, an inhibitor of prostacyclin synthesis from arachidonic acid, indicating that endogenously generated prostacyclin may modulate PAF synthesis. The potential to synthesize PAF, a unique lipid autocoid that stimulates the activation of both platelets and polymorphonuclear leukocytes, suggests that endothelial cells can directly influence the activity of these circulating effector cells. This biologic potential may be important in the interaction of the endothelium with circulating blood cells in physiologic conditions and in syndromes of vascular injury.