Effects of pyrrophenone, an inhibitor of group IVA phospholipase A2, on eicosanoid and PAF biosynthesis in human neutrophils.
OBJECTIVE
The biosynthesis of leukotrienes (LT) and platelet-activating factor (PAF) involves the release of their respective precursors, arachidonic acid (AA) and lyso-PAF by the group IVA PLA2 (cPLA2alpha). This paper aims at characterizing the inhibitory properties of the cPLA2alpha inhibitor pyrrophenone on eicosanoids and PAF in human neutrophils (PMN).


METHODS
Freshly isolated human PMN were activated with physiological and pharmacological agents (fMLP, PAF, exogenous AA, A23187 and thapsigargin) in presence and absence of the cPLA2alpha inhibitor pyrrophenone and biosynthesis of LT, PAF, and PGE2 was measured.


RESULTS
Pyrrophenone potently inhibited LT, PGE2 and PAF biosynthesis in PMN with IC50s in the range of 1-20 nM. These inhibitory effects of pyrrophenone were specific (the consequence of substrate deprivation), as shown by the reversal of inhibition by exogenous AA and lyso-PAF. Comparative assessment of pyrrophenone, methyl-arachidonoyl-fluoro-phosphonate (MAFP) and arachidonoyl-trifluoromethylketone (AACOCF3) demonstrated that pyrrophenone was more specific and 100-fold more potent than MAFP and AACOCF3 for the inhibition of LT biosynthesis in A23187-activated PMN. The inhibitory effect of pyrrophenone on LT biosynthesis was reversible as LT biosynthesis was recovered when pyrrophenone-treated PMN were washed with autologous plasma. No alteration of phospholipase D (PLD) activity in fMLP-activated PMN was observed with up to 10 microM pyrrophenone, suggesting that the cPLA2alpha inhibitor does not directly inhibit PLD.


CONCLUSIONS
Pyrrophenone is a more potent and specific cPLA2alpha inhibitor than MAFP and AACOCF3 and represents an excellent pharmacological tool to investigate the biosynthesis and the biological roles of eicosanoids and PAF.
Flamand
N
N
Département d'anatomic et de physiologie, Centre de Recherche en Rhumatologie et Immunologie, Centre de recherche du CHUQ (CHUL), Faculté de Médecine, Université Laval, Québec, Canada.
Picard
S
S
Lemieux
L
L
Pouliot
M
M
Bourgoin
S G
SG
Borgeat
P
P
D001095:Arachidonic Acids / Q000494:pharmacology; D015232:Dinoprostone / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D015777:Eicosanoids / Q000096:biosynthesis*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D054513:Group IV Phospholipases A2; D006801:Humans; D066298:In Vitro Techniques; D015289:Leukotrienes / Q000096:biosynthesis; D009504:Neutrophils / Q000187:drug effects* / Q000378:metabolism; D063065:Organophosphonates / Q000494:pharmacology; D010739:Phospholipase D / Q000037:antagonists & inhibitors / Q000378:metabolism; D010741:Phospholipases A / Q000037:antagonists & inhibitors* / Q000378:metabolism; D054467:Phospholipases A2; D010972:Platelet Activating Factor / Q000096:biosynthesis*; D011759:Pyrrolidines / Q000494:pharmacology*
D001095:Arachidonic Acids; D015777:Eicosanoids; D004791:Enzyme Inhibitors; D015289:Leukotrienes; D063065:Organophosphonates; D010972:Platelet Activating Factor; D011759:Pyrrolidines; C104810:methyl arachidonylfluorophosphonate; C455981:pyrrophenone; C081565:arachidonyltrifluoromethane; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; D054467:Phospholipases A2; D010739:Phospholipase D; D015232:Dinoprostone