title: Endothelial inositol phosphate generation and prostacyclin production in response to G-protein activation by AlF4-.
abstract: In order to elucidate the role of guanine-nucleotide-binding proteins (G-proteins) in endothelial prostacyclin (PGI2) production, human umbilical vein endothelial cells, prelabelled with either [3H]inositol or [3H]arachidonic acid, were stimulated with the non-specific G-protein activator aluminium fluoride (AlF4-). AlF4- caused a dose- and time-dependent generation of inositol phosphates, release of arachidonic acid and production of PGI2. The curves for the three events were similar. When the cells were stimulated in low extracellular calcium (60 nM), they released [3H]arachidonic acid and produced PGI2, but depleting the intracellular Ca2+ stores by pretreatment with the Ca2+ ionophore A23187 totally inhibited both events, although the cells still responded when extracellular Ca2+ was added. The Ca2+ ionophore did not inhibit the generation of inositol phosphates in cells maintained at low extracellular Ca2+. Pertussis toxin pretreatment (14 h) altered neither inositol phosphate nor PGI2 production in response to AlF4-. To investigate the functional role of the diacylglycerol/protein kinase C arm of the phosphoinositide system, the cells were pretreated with the protein kinase C activator 12-O-tetradecanoylphorbol 13-acetate (TPA) or the protein kinase C inhibitor 1-(5-isoquinolinylsulphonyl)-2-methylpiperazine (H7). TPA inhibited the AlF4(-)-induced inositol phosphate generation but stimulated both the release of arachidonic acid and the production of PGI2. H7 had opposite effects both on inositol phosphate generation and on PGI2 production. These results suggest that AlF4(-)-induced PGI2 production is mediated by a pertussis-toxin-insensitive G-protein which activates the phosphoinositide second messenger system. This production of PGI2 can be modulated by protein kinase C activation, both at the level of inositol phosphate generation and at the level of arachidonic acid release.
authors.affiliation: Department of Pharmacology, University of Iceland, Reykjavik.
authors.forename: M K
authors.initials: MK
authors.lastname: Magnússon
authors.forename: H
authors.initials: H
authors.lastname: Halldórsson
authors.forename: M
authors.initials: M
authors.lastname: Kjeld
authors.forename: G
authors.initials: G
authors.lastname: Thorgeirsson
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D015121:6-Ketoprostaglandin F1 alpha / Q000096:biosynthesis; D000535:Aluminum / Q000494:pharmacology*; D017607:Aluminum Compounds; D001095:Arachidonic Acids / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism*; D011464:Epoprostenol / Q000096:biosynthesis*; D005260:Female; D005459:Fluorides; D005461:Fluorine / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism*; D007546:Isoquinolines / Q000494:pharmacology; D007700:Kinetics; D037342:Pertussis Toxin; D010879:Piperazines / Q000494:pharmacology; D011247:Pregnancy; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D012969:Sodium Fluoride / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014471:Umbilical Veins; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
chemical_list: D017607:Aluminum Compounds; D001095:Arachidonic Acids; D007295:Inositol Phosphates; D007546:Isoquinolines; D010879:Piperazines; D010566:Virulence Factors, Bordetella; C050992:tetrafluoroaluminate; D005461:Fluorine; D000001:Calcimycin; D015121:6-Ketoprostaglandin F1 alpha; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D012969:Sodium Fluoride; D000535:Aluminum; D011464:Epoprostenol; D037342:Pertussis Toxin; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D013755:Tetradecanoylphorbol Acetate; D005459:Fluorides; D002118:Calcium