title: Activation of group VI phospholipase A2 isoforms in cardiac endothelial cells.
abstract: The endothelium comprises a cellular barrier between the circulation and tissues. We have previously shown that activation of protease-activated receptor 1 (PAR-1) and PAR-2 on the surface of human coronary artery endothelial cells by tryptase or thrombin increases group VIA phospholipase A(2) (iPLA(2)β) activity and results in production of multiple phospholipid-derived inflammatory metabolites. We isolated cardiac endothelial cells from hearts of iPLA(2)β-knockout (iPLA(2)β-KO) and wild-type (WT) mice and measured arachidonic acid (AA), prostaglandin I(2) (PGI(2)), and platelet-activating factor (PAF) production in response to PAR stimulation. Thrombin (0.1 IU/ml) or tryptase (20 ng/ml) stimulation of WT endothelial cells rapidly increased AA and PGI(2) release and increased PAF production. Selective inhibition of iPLA(2)β with (S)-bromoenol lactone (5 μM, 10 min) completely inhibited thrombin- and tryptase-stimulated responses. Thrombin or tryptase stimulation of iPLA(2)β-KO endothelial cells did not result in significant PAF production and inhibited AA and PGI(2) release. Stimulation of cardiac endothelial cells from group VIB (iPLA(2)γ)-KO mice increased PAF production to levels similar to those of WT cells but significantly attenuated PGI(2) release. These results indicate that cardiac endothelial cell PAF production is dependent on iPLA(2)β activation and that both iPLA(2)β and iPLA(2)γ may be involved in PGI(2) release.
authors.affiliation: Department of Pathology, Saint Louis University School of Medicine, Missouri, USA.
authors.forename: Janhavi
authors.initials: J
authors.lastname: Sharma
authors.forename: John
authors.initials: J
authors.lastname: Turk
authors.forename: David J
authors.initials: DJ
authors.lastname: Mancuso
authors.forename: Harold F
authors.initials: HF
authors.lastname: Sims
authors.forename: Richard W
authors.initials: RW
authors.lastname: Gross
authors.forename: Jane
authors.initials: J
authors.lastname: McHowat
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology*; D042783:Endothelial Cells / Q000166:cytology / Q000201:enzymology*; D004789:Enzyme Activation; D011464:Epoprostenol / Q000378:metabolism; D054522:Group VI Phospholipases A2 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout
chemical_list: D007527:Isoenzymes; D016718:Arachidonic Acid; D011464:Epoprostenol; D054522:Group VI Phospholipases A2