RT Journal Article
SR Electronic
T1 Inhibition of Platelet-Activating Factor (PAF) Acetylhydrolase by Methyl Arachidonyl Fluorophosphonate Potentiates PAF Synthesis in Thrombin-Stimulated Human Coronary Artery Endothelial Cells
JF Journal of Pharmacology and Experimental Therapeutics
JO J Pharmacol Exp Ther
FD American Society for Pharmacology and Experimental Therapeutics
SP 1163
OP 1170
DO 10.1124/jpet.103.055392
VO 307
IS 3
A1 Kell, Pamela J.
A1 Creer, Michael H.
A1 Crown, Kimberley N.
A1 Wirsig, Karin
A1 McHowat, Jane
YR 2003
UL http://jpet.aspetjournals.org/content/307/3/1163.abstract
AB We have previously demonstrated that thrombin stimulation of endothelial cells results in increased membrane-associated, Ca2+-independent phospholipase A2 (iPLA2) activity, accelerated hydrolysis of membrane plasmalogen phospholipids, and production of several biologically active phospholipid metabolites, including prostacyclin and platelet-activating factor (PAF) that is abolished by pretreatment with the iPLA2-selective inhibitor bromoenol lactone. This study was designed to further investigate the role of alternative PLA2 inhibitors, including methyl arachidonyl fluorophosphonate (MAFP, an inhibitor of cytosolic PLA2 isoforms), on phospholipid turnover and PAF production from thrombin-stimulated human coronary artery endothelial cells (HCAECs). Paradoxically, pretreatment of HCAEC with MAFP (5–25 μM) resulted in a significant increase in PAF production in both unstimulated and thrombin-stimulated cells that was found to be a direct result of inhibition of PAF acetylhydrolase (PAF-AH) activity. Pretreatment with MAFP did not significantly inhibit HCAEC PLA2 activity, possibly due to the localization of PLA2 activity in the membrane fraction rather than the cytosol. Bromoenol lactone did not inhibit PAF-AH activity, even at concentrations as high as 20 μM. We conclude that MAFP augments thrombin-stimulated PAF production by inhibition of PAF catabolism without affecting membrane-associated iPLA2 activity. The American Society for Pharmacology and Experimental Therapeutics