title: Electrophysiological and functional effects of sphingosine-1-phosphate in mouse ventricular fibroblasts.
abstract: The aim of this study was to characterize the effects of sphingosine-1-phosphate (S1P) on cardiac ventricular fibroblasts. Impacts of S1P on fibroblast excitability, cell migration, proliferation and secretion were characterized. The patch-clamp technique in the whole-cell configuration was used to study the S1P-induced current from mouse ventricular fibroblasts. The expression level of the S1P receptor during cell culture duration was evaluated by western-blot. Fibroblast proliferation and migration were quantified using the methylene blue assay and the Boyden chamber technique, respectively. Finally, fibroblast secretion properties were estimated by quantification of the IL-6 and collagen levels using ELISA and SIRCOL collagen assays, respectively. We found that S1P activated SUR2/Kir6.1 channel and that this effect was sensitive to specific inhibition of the S1P receptor of type 3 (S1P3R). In contrast, S1P1R receptor inhibition had no effect. Moreover, the S1P-induced current increased with cell culture duration whereas S1P3R expression level remained constant. The activation of SUR2/Kir6.1 channel by S1P via S1P3R stimulated cell proliferation and decreased IL-6 and collagen secretions. S1P also stimulated fibroblast migration via S1P3R but independently from SUR2/Kir6.1 channel activation. This study demonstrates that S1P, via S1P3R, affects cardiac ventricular fibroblasts function independently or through activation of SUR2/Kir6.1 channel. The latter effect occurs after fibroblasts differentiate into myofibroblasts, opening a new potential therapeutic strategy to modulate fibrosis after cardiac physiopathological injury.
authors.affiliation: UMR CNRS/Université de Poitiers No. 6187, Pôle Biologie Santé Bât B36, BP 633, 1 rue Georges Bonnet, 86022 Poitiers, France.
authors.forename: Najate
authors.initials: N
authors.lastname: Benamer
authors.forename: Nassim
authors.initials: N
authors.lastname: Fares
authors.forename: Patrick
authors.initials: P
authors.lastname: Bois
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Faivre
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D005905:Glyburide / Q000494:pharmacology; D006352:Heart Ventricles / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism; D054086:KATP Channels / Q000378:metabolism; D008246:Lysophospholipids / Q000494:pharmacology / Q000502:physiology*; D051379:Mice; D018408:Patch-Clamp Techniques; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism; D011955:Receptors, Drug / Q000378:metabolism; D049349:Receptors, Lysosphingolipid / Q000819:agonists / Q000502:physiology*; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000502:physiology; D000081025:Sphingosine-1-Phosphate Receptors; D064233:Sulfonylurea Receptors; D016276:Ventricular Function / Q000187:drug effects / Q000502:physiology*
chemical_list: D018528:ATP-Binding Cassette Transporters; D015850:Interleukin-6; D054086:KATP Channels; D008246:Lysophospholipids; D024661:Potassium Channels, Inwardly Rectifying; D011955:Receptors, Drug; D049349:Receptors, Lysosphingolipid; C501285:S1pr3 protein, mouse; D000081025:Sphingosine-1-Phosphate Receptors; D064233:Sulfonylurea Receptors; C092520:uK-ATP-1 potassium channel; C060506:sphingosine 1-phosphate; D003094:Collagen; D013110:Sphingosine; D005905:Glyburide