Regulation of leukotriene and platelet-activating factor synthesis in human alveolar macrophages

Abstract

It has been suggested that phospholipase A 2 (PLA 2) contributes to the regulation of leukotriene (LT) and platelet-activating factor (PAF) synthesis by controlling the release of their precursors, arachidonic acid (AA) and lysophosphatidylcholine (IysoPC), from membrane phospholipids. In rat alveolar macrophages (AMs), PLA 2 appears to have a major role in LT synthesis but a more limited role in PAF synthesis. The present study was designed to define the role of PLA 2 in LT and PAF synthesis in human AMs and determine whether differences exist between AMs obtained from normal subjects and those from patients with asthma. In the normal subjects, the calcium ionophore A23187 (Cal) increased AM PAF synthesis (percent incorporation of tritiated acetate) by 135% ( p < 0.01) and LTB 4 synthesis 88-fold ( p < 0.001). Phorbol myristate acetate (PMA) had little effect alone, but it had a synergistic effect with Cal, increasing PAF synthesis by 466% and LTB 4 synthesis to 229-fold above the control values ( p < 0.001 for both). Ro 25-4331, a combined cytosolic (c) and secretary (s) PLA 2 inhibitor, had little effect on the Cal-stimulated PAF synthesis, but it completely blocked the effect of PMA. It also blocked the Cal− and Cal+PMAstimulated LTB 4 synthesis. AACOCF 3, a cPLA 2 inhibitor, had no effect on either Cal or Cal+PMA-stimulated PAF synthesis. It reduced LTB 4 synthesis, but it did so less effectively than Ro 25-4331. CoA-independent transacylase (CoAl-TA) activity in the AMs increased after stimulation and exposure to Ro 25-4331. SK&F 45905, a CoAl-TA inhibitor, reduced stimulated PAF synthesis by 30% to 40%. Patients with asthma had similar results except that cPLA 2 had a greater role in stimulated LTB 4 synthesis. These data indicate that PLA 2 plays a direct role in human AM LT synthesis; both the cytosolic and secretary forms contribute to LT synthesis; PLA 2 appears to have a more limited role in PAF synthesis, although it mediates the synergistic effect of PMA, probably via sPLA 2 and CoAl-TA contributes to PAF synthesis during PLA 2 inhibition. With the exception of the greater role for cPLA 2 in stimulated LTB 4 synthesis in the patients with asthma, the contributions of PLA 2 and CoAl-TA to AM LT and IPAF synthesis appear to be similar in normal subjects and patients with asthma.