Metabolism of platelet-activating factor in the guinea-pig heart

Abstract

Platelet-activating factor (PAF; 1-alkyl-2-acetyl-glycerophosphocholine) is a biologically active phospholipid which is synthesized by a variety of blood cells and organ systems. PAF exerts many effects on the cardiovascular system including hypotension, depression of myocardial contractility and coronary constriction. The present study has examined the capacity of the guinea-pig heart to regulate the levels of exogenous PAF in two different models: isolated perfused heart and isolated ventricular myocytes. In the first model, isolated hearts were perfused with labeled PAF (10 −10 m) in a recirculating manner at flow rates of 15 ml/min (normal flow perfusion; NFP) and 2 ml/min (low flow perfusion, LFP). Exogenously provided PAF appeared in the tissue in a time-dependent manner. The rate of extraction of PAF was higher during LFP than during NFP. PAF was metabolized by the heart to two major products, lyso-PAF and 1-alkyl-2-acyl-sn-glycero-3-phosphocholine (1-alkyl-2-acyl-GPC). Lyso-PAF was found primarily in the perfusion buffer while both lyso-PAF and 1-alkyl-2-acyl-GPC were detected in the tissue. No qualitative difference in the metabolic products derived from PAF catabolism was observed between hearts undergoing NFP and LFP. Acetyl hydrolase activity was detected in the perfusion fluid at both flow rates, probably accounting for the formation of lyso-PAF in the perfusate. However, perfusion fluid from LFP contained a higher acetyl hydrolase activity per μg of protein as compared to fluid from NFP. Isolated ventricular myocytes incubated with labeled PAF (3 × 10 −9 m) also converted it to 1-alkyl-2-acyl-GPC. Kinetic experiments suggested that PAF was initially deacetylated to form lyso-PAF and that this intermediate was then rapidly reacylated with a fatty acyl moiety at the sn-2 position. HPLC analysis of the fatty acids inserted at the sn-2 position of 1-alkyl-2-acyl-GPC revealed that the myocytes reacylated lyso-PAF predominantly with arachidonic acid. These data indicate that the guinea-pig heart may regulate PAF levels by at least two mechanisms: (1) it may release acetyl hydrolase into the vascular compartment, particularly under low flow conditions; and (2) the ventricular myocyte has the capacity to take up PAF and catabolize it to inactive products.