title: Gene silencing of endothelial von Willebrand Factor attenuates angiotensin II-induced endothelin-1 expression in porcine aortic endothelial cells.
abstract: Expression of endothelin (ET)-1 is increased in endothelial cells exposed to angiotensin II (Ang II), leading to endothelial dysfunction and cardiovascular disorders. Since von Willebrand Factor (vWF) blockade improves endothelial function in coronary patients, we hypothesized that targeting endothelial vWF with short interference RNA (siRNA) prevents Ang II-induced ET-1 upregulation. Nearly 65 ± 2% silencing of vWF in porcine aortic endothelial cells (PAOECs) was achieved with vWF-specific siRNA without affecting cell viability and growth. While showing ET-1 similar to wild type cells at rest, vWF-silenced cells did not present ET-1 upregulation during exposure to Ang II (100 nM/24 h), preserving levels of endothelial nitric oxide synthase activity similar to wild type. vWF silencing prevented AngII-induced increase in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) activity and superoxide anion (O2-) levels, known triggers of ET-1 expression. Moreover, no increase in O2- or ET-1 levels was found in silenced cells treated with AngII or NOX-agonist phorbol ester (PMA 5 nM/48 h). Finally, vWF was required for overexpression of NOX4 and NOX2 in response to AngII and PMA. In conclusion, endothelial vWF knockdown prevented Ang II-induced ET-1 upregulation through attenuation of NOX-mediated O2- production. Our findings reveal a new role of vWF in preventing of Ang II-induced endothelial dysfunction.
authors.affiliation: Laboratory of Medical Sciences, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.
authors.forename: Anar
authors.initials: A
authors.lastname: Dushpanova
authors.affiliation: Laboratory of Medical Sciences, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Agostini
authors.affiliation: Laboratory of Medical Sciences, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.|Fondazione CNR/Regione Toscana "G. Monasterio", Pisa, Italy.
authors.forename: Enrica
authors.initials: E
authors.lastname: Ciofini
authors.affiliation: Institute of Clinical Physiology, National Council of Research, Pisa, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Cabiati
authors.affiliation: Laboratory of Medical Sciences, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Casieri
authors.affiliation: Laboratory of Medical Sciences, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Matteucci
authors.affiliation: Institute of Clinical Physiology, National Council of Research, Pisa, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Del Ry
authors.affiliation: Laboratory of Medical Sciences, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.|Fondazione CNR/Regione Toscana "G. Monasterio", Pisa, Italy.
authors.forename: Aldo
authors.initials: A
authors.lastname: Clerico
authors.affiliation: Fondazione CNR/Regione Toscana "G. Monasterio", Pisa, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Berti
authors.affiliation: Laboratory of Medical Sciences, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.|Fondazione CNR/Regione Toscana "G. Monasterio", Pisa, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Lionetti
mesh_terms: D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology*; D019332:Endothelin-1 / Q000378:metabolism*; D020868:Gene Silencing; D013552:Swine; D014841:von Willebrand Factor / Q000235:genetics / Q000378:metabolism*
chemical_list: D019332:Endothelin-1; D014841:von Willebrand Factor; D000804:Angiotensin II