title: Interleukin-1beta stimulates platelet-activating factor production in U-937 cells modulating both its biosynthetic and catabolic enzymes.
abstract: Interleukin-1beta (IL-1β) is a potent agonist of platelet-activating factor (PAF) synthesis. The monocyte-derived PAF may amplify the inflammatory and thrombotic processes. The IL-1β-induced enzymatic alterations leading to increased PAF synthesis are ill-defined. In the present study the last enzymatic activities of the remodeling (acetyl-CoA:lyso-PAF acetyltransferase) and de novo (DTT-insensitive CDP-choline:1-alkyl-2-acetyl-sn-glycerol cholinephosphotransferase) biosynthetic routes of PAF and its main catabolic enzyme, PAF acetylhydrolase, along with the intracellular and extracellular PAF levels were determined in homogenates and medium of U-937 after their stimulation with recombinant IL-1β. IL-1β at 2.5ng/mL induced an early (0.5-3h) and a late (12h) elevation of intracellular PAF levels (2-fold). Only a small portion of intracellular PAF (∼10%) was released to the extracellular medium. IL-1β increased lyso-PAF acetyltrasnferase activity which was peaked at 3h and kept elevated till 12h. A rapid 1.5-fold increase of cholinephosphotransferase activity was observed in IL-1β stimulated cells. Finally, a transient stimulation of intracellular PAF-AH was induced by IL-1β at 3h while incubation of U-937 with the PAF acetylhydrolase inhibitor pefabloc in the presence or absence of IL-1β led to a strong sustained increase of intracellular PAF levels. In conclusion, both biosynthetic routes of PAF, along with its degradation can be modulated by IL-1β in a time-specific manner. The inhibition of PAF acetylhydrolase strongly augments PAF's intracellular levels implying its crucial role for the regulation of cellular PAF. The regulation of PAF's enzymatic machinery under inflammatory conditions is more complicated than we thought to be.
authors.affiliation: Faculty of Chemistry, National and Kapodistrian University, Athens, Greece.
authors.forename: Ioanna C
authors.initials: IC
authors.lastname: Vlachogianni
authors.forename: Tzortzis
authors.initials: T
authors.lastname: Nomikos
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Fragopoulou
authors.forename: George M
authors.initials: GM
authors.lastname: Stamatakis
authors.forename: Haralambos C
authors.initials: HC
authors.lastname: Karantonis
authors.forename: Smaragdi
authors.initials: S
authors.lastname: Antonopoulou
authors.forename: Constantinos A
authors.initials: CA
authors.lastname: Demopoulos
mesh_terms: D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase / Q000037:antagonists & inhibitors; D000123:Acetyltransferases / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism; D045744:Cell Line, Tumor; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism*; D010972:Platelet Activating Factor / Q000096:biosynthesis* / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000378:metabolism; D013450:Sulfones / Q000378:metabolism
chemical_list: D053583:Interleukin-1beta; D010972:Platelet Activating Factor; D015842:Serine Proteinase Inhibitors; D013450:Sulfones; C002010:4-(2-aminoethyl)benzenesulfonylfluoride; D000123:Acetyltransferases; C046035:1-alkylglycerophosphocholine acetyltransferase; D002798:Diacylglycerol Cholinephosphotransferase; D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase