title: The active form of vitamin D, calcitriol, induces a complex dual upregulation of endothelin and nitric oxide in cultured endothelial cells.
abstract: Despite the presence of vitamin D receptor (VDR) in endothelial cells, the effect of vitamin D on endothelial function is unknown. An unbalanced production of vasoactive endothelial factors such as nitric oxide (NO) or endothelin-1 (ET-1) results in endothelial dysfunction, which can alter the normal cardiovascular function. Present experiments were devoted to assess the effect of active vitamin D (calcitriol) on the synthesis of endothelial vasoactive factors. The results were that, in cells, calcitriol increased ET-1 and NO productions, which were measured by ELISA and fluorimetric assay, respectively. Calcitriol also increased endothelin-converting enzyme-1 (ECE-1) and endothelial-nitric oxide synthase (eNOS) activities, their mRNA (qPCR), their protein expressions (Western-blot), and their promoter activities (transfection assays). Calcitriol did not change prepro-ET-1 mRNA. The effect was specific to VDR activation because when VDR was silenced by siRNA, the observed effects disappeared. Mechanisms involved in each upregulation differed. ECE-1 upregulation depended on AP-1 activation, whereas eNOS upregulation depended directly on VDR activation. To evaluate the in vivo consequences of acute calcitriol treatment, normal Wistar rats were treated with a single ip injection of 400 ng/kg calcitriol and euthanized 24 h later. Results confirmed those observed in cells, that production and expression of both factors were increased by calcitriol. Besides, calcitriol-treated rats showed a slight rise in mean blood pressure, which decreased when pretreated with FR-901533, an ECE-1 antagonist. We conclude that calcitriol increases the synthesis of both ET-1 and NO in endothelial cells. However, the ET-1 upregulation seems to be biologically more relevant, as animals acutely treated with calcitriol show slight increases in blood pressure.
authors.affiliation: Research Unit and Nephrology Section, Fundación para la Investigación Biomédica del Hospital Universitario Príncipe de Asturias, Alcalá de Henares, Madrid, Spain;
authors.forename: Patricia
authors.initials: P
authors.lastname: Martínez-Miguel
authors.affiliation: IBRLleida, Hospital Universitario Arnau de Vilanova, Lérida, Spain;
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Valdivielso
authors.affiliation: Research Unit and.
authors.forename: Diana
authors.initials: D
authors.lastname: Medrano-Andrés
authors.affiliation: Servicio de Metabolismo Mineral y Óseo, Hospital Universitario Central de Asturias, Universidad de Oviedo, Oviedo, Spain; Instituto Reina Sofía de Investigación Nefrológica, Madrid, Spain; and.
authors.forename: Pablo
authors.initials: P
authors.lastname: Román-García
authors.affiliation: Physiology Department, Universidad de Alcalá, Madrid, Spain.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Cano-Peñalver
authors.affiliation: Instituto Reina Sofía de Investigación Nefrológica, Madrid, Spain; and Physiology Department, Universidad de Alcalá, Madrid, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Rodríguez-Puyol
authors.affiliation: Research Unit and Nephrology Section, Fundación para la Investigación Biomédica del Hospital Universitario Príncipe de Asturias, Alcalá de Henares, Madrid, Spain; Instituto Reina Sofía de Investigación Nefrológica, Madrid, Spain; and.
authors.forename: Diego
authors.initials: D
authors.lastname: Rodríguez-Puyol
authors.affiliation: Research Unit and Instituto Reina Sofía de Investigación Nefrológica, Madrid, Spain; and slorgil@salud.madrid.org.
authors.forename: Susana
authors.identifier: http://orcid.org/0000-0003-0045-1680
authors.initials: S
authors.lastname: López-Ongil
mesh_terms: D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000235:genetics / Q000378:metabolism; D001794:Blood Pressure / Q000187:drug effects / Q000235:genetics; D002117:Calcitriol / Q000494:pharmacology*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D000072396:Endothelin-Converting Enzymes; D016232:Endothelins / Q000378:metabolism*; D006801:Humans; D008297:Male; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D018167:Receptors, Calcitriol / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: D016232:Endothelins; D018167:Receptors, Calcitriol; C579457:VDR protein, human; D009569:Nitric Oxide; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; C000606923:ECE1 protein, human; D000072396:Endothelin-Converting Enzymes; D002117:Calcitriol
keywords: endothelin-1; endothelium; nitric oxide; vitamin D