Detection of platelet-activating factor in exudates of rats with phorbol myristate acetate-induced pleurisy

Abstract

PAF-likely activity, detected as aggregation of washed platelets, was found in the exudate of rats with pleurisy induced by phorbol myristate acetate (PMA, 1 nmol). At 30 min after the injection of PMA, 400-500 pg of PAF was detected in the pleural exudate. An extract of the exudate was made and analysed by HPLC and by rabbit platelet aggregation. The activity was characterized as that of PAF as a result of the inhibition seen with the PAF-antagonist CV-3988 and the loss of activity by treatment with phospholipase A2. Indirect evidence was previously reported in that CV-3988 suppressed the pleural fluid accumulation in the pleurisy induced by PMA. Taken together these facts it indicates that PAF could be an important mediator of acute inflammation, especially of the plasma exudation resulting from an increase in vascular permeability. Evidence that pleural cells produce PAF was also examined in an in vitro experiment. PAF was found in the supernatant and cellular fractions of the incubation mixture of the pleural cells from normal rats when stimulated by PMA (1 microM) or A23187 (5 microM), and the results indicate that the pleural cells produce PAF and release a portion of it.