title: Involvement of proteinase activated receptor-2 in the vascular response to sphingosine 1-phosphate.
abstract: S1P (sphingosine 1-phosphate) represents one of the key latest additions to the list of vasoactive substances that modulate vascular tone. PAR-2 (proteinase activated receptor-2) has been shown to be involved in cardiovascular function. In the present study, we investigated the involvement of PAR-2 in S1P-induced effect on vascular tone. The present study has been performed by using isolated mouse aortas. Both S1P and PAR-2 agonists induced endothelium-dependent vasorelaxation. L-NAME (N(G)-nitro-L-arginine methyl ester) and wortmannin abrogated the S1P-induced vasorelaxatioin, while significantly inhibiting the PAR-2-mediated effect. Either ENMD1068, a PAR-2 antagonist, or gabexate, a serine protease inhibitor, significantly inhibited S1P-induced vasorelaxation. Aortic tissues harvested from mice overexpressing PAR-2 displayed a significant increase in vascular response to S1P as opposed to PAR-2-null mice. Immunoprecipitation and immunofluorescence studies demonstrated that S1P(1) interacted with PAR-2 and co-localized with PAR-2 on the vascular endothelial surface. Furthermore, S1P administration to vascular tissues triggered PAR-2 mobilization from the plasma membrane to the perinuclear area; S1P-induced translocation of PAR-2 was abrogated when aortic rings were pre-treated with ENMD1068 or when caveolae dysfunction occurred. Similarly, experiments performed in cultured endothelial cells (human umbilical vein endothelial cells) showed a co-localization of S1P(1) and PAR2, as well as the ability of S1P to induce PAR-2 trafficking. Our results suggest that S1P induces endothelium-dependent vasorelaxation mainly through S1P(1) and involves PAR-2 transactivation.
authors.affiliation: *Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy.
authors.forename: Fiorentina
authors.initials: F
authors.lastname: Roviezzo
authors.affiliation: †Dipartimento di Medicina Sperimentale, Sezione di Farmacologia L. Donatelli, Seconda Università degli Studi di Napoli, Napoli, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: De Angelis
authors.affiliation: *Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy.
authors.forename: Luana
authors.initials: L
authors.lastname: De Gruttola
authors.affiliation: *Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Bertolino
authors.affiliation: †Dipartimento di Medicina Sperimentale, Sezione di Farmacologia L. Donatelli, Seconda Università degli Studi di Napoli, Napoli, Italy.
authors.forename: Nikol
authors.initials: N
authors.lastname: Sullo
authors.affiliation: ‡Dipartimento di Chimica, Università degli Studi della Basilicata, Pontenza, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Brancaleone
authors.affiliation: *Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy.
authors.forename: Mariarosaria
authors.initials: M
authors.lastname: Bucci
authors.affiliation: §Dipartimento di Internistica Clinica e Sperimentale, Seconda Università degli Studi di Napoli, Napoli, Italy.
authors.forename: Raffaele
authors.initials: R
authors.lastname: De Palma
authors.affiliation: †Dipartimento di Medicina Sperimentale, Sezione di Farmacologia L. Donatelli, Seconda Università degli Studi di Napoli, Napoli, Italy.
authors.forename: Konrad
authors.initials: K
authors.lastname: Urbanek
authors.affiliation: †Dipartimento di Medicina Sperimentale, Sezione di Farmacologia L. Donatelli, Seconda Università degli Studi di Napoli, Napoli, Italy.
authors.forename: Bruno
authors.initials: B
authors.lastname: D'Agostino
authors.affiliation: *Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Ianaro
authors.affiliation: *Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Sorrentino
authors.affiliation: *Dipartimento di Farmacia, Università di Napoli Federico II, Napoli, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Cirino
mesh_terms: D000730:Androstadienes; D000818:Animals; D001011:Aorta / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D015153:Blotting, Western; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D016670:Gabexate / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008246:Lysophospholipids / Q000494:pharmacology*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010879:Piperazines / Q000494:pharmacology; D021381:Protein Transport / Q000187:drug effects; D044464:Receptor, PAR-2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D049349:Receptors, Lysosphingolipid / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D014664:Vasodilation / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D016670:Gabexate; D008246:Lysophospholipids; D019331:NG-Nitroarginine Methyl Ester; D019001:Nitric Oxide Synthase; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010879:Piperazines; D044464:Receptor, PAR-2; D049349:Receptors, Lysosphingolipid; D015842:Serine Proteinase Inhibitors; D013110:Sphingosine; D000077191:Wortmannin; C518710:1-3-methylbutyryl-N4-6-aminohexanoyl-piperazine; C060506:sphingosine 1-phosphate