In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line

Autores
Cardona, Paula; Palacios, Daniel; Alvarez, Anny; Damiano, Alicia Ermelinda; García Robles, Reggie; Ayala Ramírez, Paola
Año de publicación
2026
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Introduction. The placenta is a transient organ essential for fetal development. Abnormal placental development can lead to obstetric syndromes, such as preeclampsia (PE) and fetal growth restriction (FGR). Physiological hypoxia activates intracellular pathways via hypoxia-inducible transcription factors (HIFs), which are crucial for placentation. This study uses an in vitro model to investigate the role of oxygen concentration in the expression of thrombomodulin (THBD), a protein involved in placental hemostasis. Methods. Swan 71, a first-trimester trophoblast cell line, was cultured under normoxic (21 % O2) and physiological hypoxic (2 and 8 % O2) conditions. Hypoxia/reoxygenation assays were also performed. To assess potential modulation of THBD expression by HIF1α, cobalt chloride (CoCl2) was used to stabilize HIF-1α, while zinc chloride (ZnCl2) inhibited its expression. RT-qPCR and Western blot analyses were performed to quantify THBD and KLF4 expression. Results. Significant increases in THBD mRNA and protein levels were observed under 2 % O2 conditions (p = 0.03). CoCl2 treatment further enhanced THBD and KLF4 expression (p < 0.01 in both experiments), while ZnCl2 inhibited these effects (p < 0.01). This suggests a key role for HIF-1α in THBD regulation. However, in silico analysis did not identify any direct hypoxia response elements within the human THBD gene, suggesting the presence of an indirect regulatory mechanism potentially involving KLF4. Discussion. These results suggest that physiological hypoxia positively regulates THBD expression through HIF-1α stabilization and KLF4-mediation. These findings provide insight into the molecular mechanisms of placental adaptation to hypoxia and are relevant for understanding pregnancy complications like PE and intrauterine growth restriction (IUGR).
Fil: Cardona, Paula. Pontificia Universidad Javeriana; Colombia
Fil: Palacios, Daniel. Pontificia Universidad Javeriana; Colombia
Fil: Alvarez, Anny. Pontificia Universidad Javeriana; Colombia
Fil: Damiano, Alicia Ermelinda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Ciencias Biológicas. Cátedra de Biología Celular y Molecular; Argentina
Fil: García Robles, Reggie. Pontificia Universidad Javeriana; Colombia
Fil: Ayala Ramírez, Paola. Pontificia Universidad Javeriana; Colombia
Materia
HYPOXIA
PLACENTA
KRUPPEL-LIKE FACTOR 4
HYPOXIA-INDUCIBLE FACTOR 1
PREGNANCY COMPLICATIONS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/289914
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/289914
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell LineCardona, PaulaPalacios, DanielAlvarez, AnnyDamiano, Alicia ErmelindaGarcía Robles, ReggieAyala Ramírez, PaolaHYPOXIAPLACENTAKRUPPEL-LIKE FACTOR 4HYPOXIA-INDUCIBLE FACTOR 1PREGNANCY COMPLICATIONShttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Introduction. The placenta is a transient organ essential for fetal development. Abnormal placental development can lead to obstetric syndromes, such as preeclampsia (PE) and fetal growth restriction (FGR). Physiological hypoxia activates intracellular pathways via hypoxia-inducible transcription factors (HIFs), which are crucial for placentation. This study uses an in vitro model to investigate the role of oxygen concentration in the expression of thrombomodulin (THBD), a protein involved in placental hemostasis. Methods. Swan 71, a first-trimester trophoblast cell line, was cultured under normoxic (21 % O2) and physiological hypoxic (2 and 8 % O2) conditions. Hypoxia/reoxygenation assays were also performed. To assess potential modulation of THBD expression by HIF1α, cobalt chloride (CoCl2) was used to stabilize HIF-1α, while zinc chloride (ZnCl2) inhibited its expression. RT-qPCR and Western blot analyses were performed to quantify THBD and KLF4 expression. Results. Significant increases in THBD mRNA and protein levels were observed under 2 % O2 conditions (p = 0.03). CoCl2 treatment further enhanced THBD and KLF4 expression (p < 0.01 in both experiments), while ZnCl2 inhibited these effects (p < 0.01). This suggests a key role for HIF-1α in THBD regulation. However, in silico analysis did not identify any direct hypoxia response elements within the human THBD gene, suggesting the presence of an indirect regulatory mechanism potentially involving KLF4. Discussion. These results suggest that physiological hypoxia positively regulates THBD expression through HIF-1α stabilization and KLF4-mediation. These findings provide insight into the molecular mechanisms of placental adaptation to hypoxia and are relevant for understanding pregnancy complications like PE and intrauterine growth restriction (IUGR).Fil: Cardona, Paula. Pontificia Universidad Javeriana; ColombiaFil: Palacios, Daniel. Pontificia Universidad Javeriana; ColombiaFil: Alvarez, Anny. Pontificia Universidad Javeriana; ColombiaFil: Damiano, Alicia Ermelinda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Ciencias Biológicas. Cátedra de Biología Celular y Molecular; ArgentinaFil: García Robles, Reggie. Pontificia Universidad Javeriana; ColombiaFil: Ayala Ramírez, Paola. Pontificia Universidad Javeriana; ColombiaElsevier Science Inc.2026-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/289914Cardona, Paula; Palacios, Daniel; Alvarez, Anny; Damiano, Alicia Ermelinda; García Robles, Reggie; et al.; In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line; Elsevier Science Inc.; Archives Of Medical Research (eletronico); 57; 4; 6-2026; 1-130188-4409CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0188440925001523info:eu-repo/semantics/altIdentifier/doi/10.1016/j.arcmed.2025.103332info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-08-25T15:04:34Zoai:ri.conicet.gov.ar:11336/289914instacron: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:34982026-08-25 15:04:34.562CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
title In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
spellingShingle In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
Cardona, Paula
HYPOXIA
PLACENTA
KRUPPEL-LIKE FACTOR 4
HYPOXIA-INDUCIBLE FACTOR 1
PREGNANCY COMPLICATIONS
title_short In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
title_full In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
title_fullStr In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
title_full_unstemmed In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
title_sort In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line
dc.creator.none.fl_str_mv Cardona, Paula
Palacios, Daniel
Alvarez, Anny
Damiano, Alicia Ermelinda
García Robles, Reggie
Ayala Ramírez, Paola
author Cardona, Paula
author_facet Cardona, Paula
Palacios, Daniel
Alvarez, Anny
Damiano, Alicia Ermelinda
García Robles, Reggie
Ayala Ramírez, Paola
author_role author
author2 Palacios, Daniel
Alvarez, Anny
Damiano, Alicia Ermelinda
García Robles, Reggie
Ayala Ramírez, Paola
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv HYPOXIA
PLACENTA
KRUPPEL-LIKE FACTOR 4
HYPOXIA-INDUCIBLE FACTOR 1
PREGNANCY COMPLICATIONS
topic HYPOXIA
PLACENTA
KRUPPEL-LIKE FACTOR 4
HYPOXIA-INDUCIBLE FACTOR 1
PREGNANCY COMPLICATIONS
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Introduction. The placenta is a transient organ essential for fetal development. Abnormal placental development can lead to obstetric syndromes, such as preeclampsia (PE) and fetal growth restriction (FGR). Physiological hypoxia activates intracellular pathways via hypoxia-inducible transcription factors (HIFs), which are crucial for placentation. This study uses an in vitro model to investigate the role of oxygen concentration in the expression of thrombomodulin (THBD), a protein involved in placental hemostasis. Methods. Swan 71, a first-trimester trophoblast cell line, was cultured under normoxic (21 % O2) and physiological hypoxic (2 and 8 % O2) conditions. Hypoxia/reoxygenation assays were also performed. To assess potential modulation of THBD expression by HIF1α, cobalt chloride (CoCl2) was used to stabilize HIF-1α, while zinc chloride (ZnCl2) inhibited its expression. RT-qPCR and Western blot analyses were performed to quantify THBD and KLF4 expression. Results. Significant increases in THBD mRNA and protein levels were observed under 2 % O2 conditions (p = 0.03). CoCl2 treatment further enhanced THBD and KLF4 expression (p < 0.01 in both experiments), while ZnCl2 inhibited these effects (p < 0.01). This suggests a key role for HIF-1α in THBD regulation. However, in silico analysis did not identify any direct hypoxia response elements within the human THBD gene, suggesting the presence of an indirect regulatory mechanism potentially involving KLF4. Discussion. These results suggest that physiological hypoxia positively regulates THBD expression through HIF-1α stabilization and KLF4-mediation. These findings provide insight into the molecular mechanisms of placental adaptation to hypoxia and are relevant for understanding pregnancy complications like PE and intrauterine growth restriction (IUGR).
Fil: Cardona, Paula. Pontificia Universidad Javeriana; Colombia
Fil: Palacios, Daniel. Pontificia Universidad Javeriana; Colombia
Fil: Alvarez, Anny. Pontificia Universidad Javeriana; Colombia
Fil: Damiano, Alicia Ermelinda. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Fisiología y Biofísica Bernardo Houssay. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Fisiología y Biofísica Bernardo Houssay; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Ciencias Biológicas. Cátedra de Biología Celular y Molecular; Argentina
Fil: García Robles, Reggie. Pontificia Universidad Javeriana; Colombia
Fil: Ayala Ramírez, Paola. Pontificia Universidad Javeriana; Colombia
description Introduction. The placenta is a transient organ essential for fetal development. Abnormal placental development can lead to obstetric syndromes, such as preeclampsia (PE) and fetal growth restriction (FGR). Physiological hypoxia activates intracellular pathways via hypoxia-inducible transcription factors (HIFs), which are crucial for placentation. This study uses an in vitro model to investigate the role of oxygen concentration in the expression of thrombomodulin (THBD), a protein involved in placental hemostasis. Methods. Swan 71, a first-trimester trophoblast cell line, was cultured under normoxic (21 % O2) and physiological hypoxic (2 and 8 % O2) conditions. Hypoxia/reoxygenation assays were also performed. To assess potential modulation of THBD expression by HIF1α, cobalt chloride (CoCl2) was used to stabilize HIF-1α, while zinc chloride (ZnCl2) inhibited its expression. RT-qPCR and Western blot analyses were performed to quantify THBD and KLF4 expression. Results. Significant increases in THBD mRNA and protein levels were observed under 2 % O2 conditions (p = 0.03). CoCl2 treatment further enhanced THBD and KLF4 expression (p < 0.01 in both experiments), while ZnCl2 inhibited these effects (p < 0.01). This suggests a key role for HIF-1α in THBD regulation. However, in silico analysis did not identify any direct hypoxia response elements within the human THBD gene, suggesting the presence of an indirect regulatory mechanism potentially involving KLF4. Discussion. These results suggest that physiological hypoxia positively regulates THBD expression through HIF-1α stabilization and KLF4-mediation. These findings provide insight into the molecular mechanisms of placental adaptation to hypoxia and are relevant for understanding pregnancy complications like PE and intrauterine growth restriction (IUGR).
publishDate 2026
dc.date.none.fl_str_mv 2026-06
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/11336/289914
Cardona, Paula; Palacios, Daniel; Alvarez, Anny; Damiano, Alicia Ermelinda; García Robles, Reggie; et al.; In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line; Elsevier Science Inc.; Archives Of Medical Research (eletronico); 57; 4; 6-2026; 1-13
0188-4409
CONICET Digital
CONICET
url http://hdl.handle.net/11336/289914
identifier_str_mv Cardona, Paula; Palacios, Daniel; Alvarez, Anny; Damiano, Alicia Ermelinda; García Robles, Reggie; et al.; In Vitro Evaluation of the Impact of Oxygen Concentrations on Thrombomodulin Expression in a First-Trimester Trophoblast Cell Line; Elsevier Science Inc.; Archives Of Medical Research (eletronico); 57; 4; 6-2026; 1-13
0188-4409
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0188440925001523
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.arcmed.2025.103332
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science Inc.
publisher.none.fl_str_mv Elsevier Science 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 13.265058

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