VWF and ADAMTS13 behavior in estradiol-treated HUVEC

Autores
Powazniak, Yanina Paola; Kempfer, Ana Catalina; Calderazzo, Julio César; Paiva Palomino, Juvenal Hernán; Lazzari, María Ángela
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Objectives: In this study, the role of 17β-estradiol (E2) in the regulation of von Willebrand factor (VWF) and ADAMTS13 synthesis, storage, and secretion was investigated in cultured human umbilical vein endothelial cells (HUVEC).Methods: HUVEC were grown to 80-90% confluence and replaced with fresh medium containing E2 (1 nm) or vehicle for 24 h, after which the supernatant medium and cell lysates were collected to measure VWF and ADAMTS13. VWF was evaluated by VWF:Ag and multimeric analysis. ADAMTS13 was evaluated by SDS-PAGE. VWF and ADAMTS13 mRNA were quantified by real-time PCR after E2 or vehicle exposure for 18 h. A functional effect of ADAMTS13 on HUVEC VWF protein synthesis was further evaluated using a short hairpin RNA (shRNA) to knockdown the expression of endogenous ADAMTS13.Results: E2 did not increase the release or intracellular VWF levels in HUVEC. However, E2 increased the production of intracellular ADAMTS13, although there was no evidence of significant effects of their release into culture medium. Incubation of HUVEC with E2 resulted in a significantly increased expression of VWF and ADAMTS13 mRNA. ADAMTS13 gene inactivation upregulates release and intracellular VWF levels in E2-treated HUVEC.Conclusion: The results demonstrated that E2 may play a role in the regulation of VWF and ADAMTS13 gene expression and in its production in human endothelial cells. The mechanism of the protective effects of E2 on the cardiovascular system could be explained by the intracellular regulation of VWF produced by ADAMTS13.
Fil: Powazniak, Yanina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Kempfer, Ana Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Calderazzo, Julio César. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Paiva Palomino, Juvenal Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Lazzari, María Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Materia
17Β-ESTRADIOL
ADAMTS13
ENDOTHELIAL CELLS
VON WILLEBRAND FACTOR
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/92216
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/92216
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling VWF and ADAMTS13 behavior in estradiol-treated HUVECPowazniak, Yanina PaolaKempfer, Ana CatalinaCalderazzo, Julio CésarPaiva Palomino, Juvenal HernánLazzari, María Ángela17Β-ESTRADIOLADAMTS13ENDOTHELIAL CELLSVON WILLEBRAND FACTORhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Objectives: In this study, the role of 17β-estradiol (E2) in the regulation of von Willebrand factor (VWF) and ADAMTS13 synthesis, storage, and secretion was investigated in cultured human umbilical vein endothelial cells (HUVEC).Methods: HUVEC were grown to 80-90% confluence and replaced with fresh medium containing E2 (1 nm) or vehicle for 24 h, after which the supernatant medium and cell lysates were collected to measure VWF and ADAMTS13. VWF was evaluated by VWF:Ag and multimeric analysis. ADAMTS13 was evaluated by SDS-PAGE. VWF and ADAMTS13 mRNA were quantified by real-time PCR after E2 or vehicle exposure for 18 h. A functional effect of ADAMTS13 on HUVEC VWF protein synthesis was further evaluated using a short hairpin RNA (shRNA) to knockdown the expression of endogenous ADAMTS13.Results: E2 did not increase the release or intracellular VWF levels in HUVEC. However, E2 increased the production of intracellular ADAMTS13, although there was no evidence of significant effects of their release into culture medium. Incubation of HUVEC with E2 resulted in a significantly increased expression of VWF and ADAMTS13 mRNA. ADAMTS13 gene inactivation upregulates release and intracellular VWF levels in E2-treated HUVEC.Conclusion: The results demonstrated that E2 may play a role in the regulation of VWF and ADAMTS13 gene expression and in its production in human endothelial cells. The mechanism of the protective effects of E2 on the cardiovascular system could be explained by the intracellular regulation of VWF produced by ADAMTS13.Fil: Powazniak, Yanina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; ArgentinaFil: Kempfer, Ana Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; ArgentinaFil: Calderazzo, Julio César. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; ArgentinaFil: Paiva Palomino, Juvenal Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; ArgentinaFil: Lazzari, María Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; ArgentinaWiley Blackwell Publishing, Inc2011-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/92216Powazniak, Yanina Paola; Kempfer, Ana Catalina; Calderazzo, Julio César; Paiva Palomino, Juvenal Hernán; Lazzari, María Ángela; VWF and ADAMTS13 behavior in estradiol-treated HUVEC; Wiley Blackwell Publishing, Inc; European Journal Of Haematology; 86; 2; 2-2011; 140-1470902-4441CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1111/j.1600-0609.2010.01545.xinfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1600-0609.2010.01545.xinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:45:13Zoai:ri.conicet.gov.ar:11336/92216instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:45:14.131CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv VWF and ADAMTS13 behavior in estradiol-treated HUVEC
title VWF and ADAMTS13 behavior in estradiol-treated HUVEC
spellingShingle VWF and ADAMTS13 behavior in estradiol-treated HUVEC
Powazniak, Yanina Paola
17Β-ESTRADIOL
ADAMTS13
ENDOTHELIAL CELLS
VON WILLEBRAND FACTOR
title_short VWF and ADAMTS13 behavior in estradiol-treated HUVEC
title_full VWF and ADAMTS13 behavior in estradiol-treated HUVEC
title_fullStr VWF and ADAMTS13 behavior in estradiol-treated HUVEC
title_full_unstemmed VWF and ADAMTS13 behavior in estradiol-treated HUVEC
title_sort VWF and ADAMTS13 behavior in estradiol-treated HUVEC
dc.creator.none.fl_str_mv Powazniak, Yanina Paola
Kempfer, Ana Catalina
Calderazzo, Julio César
Paiva Palomino, Juvenal Hernán
Lazzari, María Ángela
author Powazniak, Yanina Paola
author_facet Powazniak, Yanina Paola
Kempfer, Ana Catalina
Calderazzo, Julio César
Paiva Palomino, Juvenal Hernán
Lazzari, María Ángela
author_role author
author2 Kempfer, Ana Catalina
Calderazzo, Julio César
Paiva Palomino, Juvenal Hernán
Lazzari, María Ángela
author2_role author
author
author
author
dc.subject.none.fl_str_mv 17Β-ESTRADIOL
ADAMTS13
ENDOTHELIAL CELLS
VON WILLEBRAND FACTOR
topic 17Β-ESTRADIOL
ADAMTS13
ENDOTHELIAL CELLS
VON WILLEBRAND FACTOR
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Objectives: In this study, the role of 17β-estradiol (E2) in the regulation of von Willebrand factor (VWF) and ADAMTS13 synthesis, storage, and secretion was investigated in cultured human umbilical vein endothelial cells (HUVEC).Methods: HUVEC were grown to 80-90% confluence and replaced with fresh medium containing E2 (1 nm) or vehicle for 24 h, after which the supernatant medium and cell lysates were collected to measure VWF and ADAMTS13. VWF was evaluated by VWF:Ag and multimeric analysis. ADAMTS13 was evaluated by SDS-PAGE. VWF and ADAMTS13 mRNA were quantified by real-time PCR after E2 or vehicle exposure for 18 h. A functional effect of ADAMTS13 on HUVEC VWF protein synthesis was further evaluated using a short hairpin RNA (shRNA) to knockdown the expression of endogenous ADAMTS13.Results: E2 did not increase the release or intracellular VWF levels in HUVEC. However, E2 increased the production of intracellular ADAMTS13, although there was no evidence of significant effects of their release into culture medium. Incubation of HUVEC with E2 resulted in a significantly increased expression of VWF and ADAMTS13 mRNA. ADAMTS13 gene inactivation upregulates release and intracellular VWF levels in E2-treated HUVEC.Conclusion: The results demonstrated that E2 may play a role in the regulation of VWF and ADAMTS13 gene expression and in its production in human endothelial cells. The mechanism of the protective effects of E2 on the cardiovascular system could be explained by the intracellular regulation of VWF produced by ADAMTS13.
Fil: Powazniak, Yanina Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Kempfer, Ana Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Calderazzo, Julio César. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Paiva Palomino, Juvenal Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
Fil: Lazzari, María Ángela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Ministerio de Ciencia, Tecnología e Innovación Productiva. Agencia Nacional de Promoción Científica y Tecnológica. Fondo para la Investigación Científica y Tecnológica; Argentina. Academia Nacional de Medicina de Buenos Aires; Argentina
description Objectives: In this study, the role of 17β-estradiol (E2) in the regulation of von Willebrand factor (VWF) and ADAMTS13 synthesis, storage, and secretion was investigated in cultured human umbilical vein endothelial cells (HUVEC).Methods: HUVEC were grown to 80-90% confluence and replaced with fresh medium containing E2 (1 nm) or vehicle for 24 h, after which the supernatant medium and cell lysates were collected to measure VWF and ADAMTS13. VWF was evaluated by VWF:Ag and multimeric analysis. ADAMTS13 was evaluated by SDS-PAGE. VWF and ADAMTS13 mRNA were quantified by real-time PCR after E2 or vehicle exposure for 18 h. A functional effect of ADAMTS13 on HUVEC VWF protein synthesis was further evaluated using a short hairpin RNA (shRNA) to knockdown the expression of endogenous ADAMTS13.Results: E2 did not increase the release or intracellular VWF levels in HUVEC. However, E2 increased the production of intracellular ADAMTS13, although there was no evidence of significant effects of their release into culture medium. Incubation of HUVEC with E2 resulted in a significantly increased expression of VWF and ADAMTS13 mRNA. ADAMTS13 gene inactivation upregulates release and intracellular VWF levels in E2-treated HUVEC.Conclusion: The results demonstrated that E2 may play a role in the regulation of VWF and ADAMTS13 gene expression and in its production in human endothelial cells. The mechanism of the protective effects of E2 on the cardiovascular system could be explained by the intracellular regulation of VWF produced by ADAMTS13.
publishDate 2011
dc.date.none.fl_str_mv 2011-02
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/92216
Powazniak, Yanina Paola; Kempfer, Ana Catalina; Calderazzo, Julio César; Paiva Palomino, Juvenal Hernán; Lazzari, María Ángela; VWF and ADAMTS13 behavior in estradiol-treated HUVEC; Wiley Blackwell Publishing, Inc; European Journal Of Haematology; 86; 2; 2-2011; 140-147
0902-4441
CONICET Digital
CONICET
url http://hdl.handle.net/11336/92216
identifier_str_mv Powazniak, Yanina Paola; Kempfer, Ana Catalina; Calderazzo, Julio César; Paiva Palomino, Juvenal Hernán; Lazzari, María Ángela; VWF and ADAMTS13 behavior in estradiol-treated HUVEC; Wiley Blackwell Publishing, Inc; European Journal Of Haematology; 86; 2; 2-2011; 140-147
0902-4441
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1111/j.1600-0609.2010.01545.x
info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1111/j.1600-0609.2010.01545.x
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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