Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod

Autores
Rocco, Rodrigo; Wainstok, Rosa; Cochon, Adriana; Gazzaniga, Silvina Noemí
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
The off-label use of imiquimod (IQ) for hemangioma treatment has shown clinical benefits. We have previously reported a selective di- rect IQ-cytotoxic effect on transformed H5V endothelial cells (EC) (hemangioma model) vs normal 1G11 EC. We observed a severe imbalance in antioxidant defense and apoptosis in H5V but not in 1G11. To further address this issue, we studied the possibility of IQ being a mitochondrial toxicant. H5V and 1G11 cells were treated with IQ (0-50 μg/mL) for 2, 4, 12 or 24 h and analyzed for reac- tive oxygen species (ROS) with DCFH2-DA probe and mitochondrial stress by MitoTrackerTM Red CMXRos fluorescence. Viability assays were performed using the standard culture medium with 5.5 mM glucose (regular) or media containing 25 mM glucose (high) or 25 mM galactose (depleted). IQ treatment increased ROS level in H5V after 2 h (35-60%; p<0.05) but in 1G11 only at 4 h (̴50%; p<0.05). Mitochondrial membrane potential in H5V cells was affected after 4 and 12 h treatment, revealed by a decreased in MitoTracker fluores- cence (≈50%; p<0.05). In contrast, 1G11 cells were unaffected and only presented a significant 30%-decrease in fluorescence after 12 h with 50 μg/mL IQ (p<0.05). Cells grown in a high glucose medium can adapt to a glycolytic phenotype. By assessing the effect of IQ in this medium, both cell lines became significantly less affected than with the regular culture medium. On the contrary, by forcing cells to respiration with galactose instead of glucose-containing medium, IQ treatment enhanced cell death in both cell lines, being fully cytotoxic for H5V (p<0.05) but leaving ≈32% 1G11 cells still alive at the high- est IQ concentrations.These results provide more evidences about the higher suscepti- bility of transformed EC to IQ, where an early ROS production and mitochondrial dysfunction drove H5V cells to death. By shifting cells towards diminished respiration in absence of glucose, we proved IQ acts as a mitochondrial toxicant in both EC lines.
Fil: Rocco, Rodrigo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Wainstok, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Cochon, Adriana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Fil: Gazzaniga, Silvina Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
LXVI Reunión anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión anual de la Sociedad Argentina de Immunología; LIII Reunión anual de la Asociación Argentina de Farmacología Experimental y XI Reunión anual de la Asociación Argentina de Nanomedicinas
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Asociación Argentina de Farmacología Experimental
Asociación Argentina de Nanomedicinas
Materia
IMIQUIMOD
HEMANGIOMA
ENDOTELIO
MITOCONDRIA
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:11336/249759
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimodRocco, RodrigoWainstok, RosaCochon, AdrianaGazzaniga, Silvina NoemíIMIQUIMODHEMANGIOMAENDOTELIOMITOCONDRIAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The off-label use of imiquimod (IQ) for hemangioma treatment has shown clinical benefits. We have previously reported a selective di- rect IQ-cytotoxic effect on transformed H5V endothelial cells (EC) (hemangioma model) vs normal 1G11 EC. We observed a severe imbalance in antioxidant defense and apoptosis in H5V but not in 1G11. To further address this issue, we studied the possibility of IQ being a mitochondrial toxicant. H5V and 1G11 cells were treated with IQ (0-50 μg/mL) for 2, 4, 12 or 24 h and analyzed for reac- tive oxygen species (ROS) with DCFH2-DA probe and mitochondrial stress by MitoTrackerTM Red CMXRos fluorescence. Viability assays were performed using the standard culture medium with 5.5 mM glucose (regular) or media containing 25 mM glucose (high) or 25 mM galactose (depleted). IQ treatment increased ROS level in H5V after 2 h (35-60%; p<0.05) but in 1G11 only at 4 h (̴50%; p<0.05). Mitochondrial membrane potential in H5V cells was affected after 4 and 12 h treatment, revealed by a decreased in MitoTracker fluores- cence (≈50%; p<0.05). In contrast, 1G11 cells were unaffected and only presented a significant 30%-decrease in fluorescence after 12 h with 50 μg/mL IQ (p<0.05). Cells grown in a high glucose medium can adapt to a glycolytic phenotype. By assessing the effect of IQ in this medium, both cell lines became significantly less affected than with the regular culture medium. On the contrary, by forcing cells to respiration with galactose instead of glucose-containing medium, IQ treatment enhanced cell death in both cell lines, being fully cytotoxic for H5V (p<0.05) but leaving ≈32% 1G11 cells still alive at the high- est IQ concentrations.These results provide more evidences about the higher suscepti- bility of transformed EC to IQ, where an early ROS production and mitochondrial dysfunction drove H5V cells to death. By shifting cells towards diminished respiration in absence of glucose, we proved IQ acts as a mitochondrial toxicant in both EC lines.Fil: Rocco, Rodrigo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Wainstok, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Cochon, Adriana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Gazzaniga, Silvina Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaLXVI Reunión anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión anual de la Sociedad Argentina de Immunología; LIII Reunión anual de la Asociación Argentina de Farmacología Experimental y XI Reunión anual de la Asociación Argentina de NanomedicinasArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de InmunologíaAsociación Argentina de Farmacología ExperimentalAsociación Argentina de NanomedicinasFundación Revista Medicina2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttp://hdl.handle.net/11336/249759Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod; LXVI Reunión anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión anual de la Sociedad Argentina de Immunología; LIII Reunión anual de la Asociación Argentina de Farmacología Experimental y XI Reunión anual de la Asociación Argentina de Nanomedicinas; Argentina; 2021; 105-105CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://medicinabuenosaires.com/revistas/vol81-21/s3/Mv81s3.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:43:18Zoai:ri.conicet.gov.ar:11336/249759instacron: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 09:43:18.657CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
title Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
spellingShingle Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
Rocco, Rodrigo
IMIQUIMOD
HEMANGIOMA
ENDOTELIO
MITOCONDRIA
title_short Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
title_full Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
title_fullStr Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
title_full_unstemmed Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
title_sort Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod
dc.creator.none.fl_str_mv Rocco, Rodrigo
Wainstok, Rosa
Cochon, Adriana
Gazzaniga, Silvina Noemí
author Rocco, Rodrigo
author_facet Rocco, Rodrigo
Wainstok, Rosa
Cochon, Adriana
Gazzaniga, Silvina Noemí
author_role author
author2 Wainstok, Rosa
Cochon, Adriana
Gazzaniga, Silvina Noemí
author2_role author
author
author
dc.subject.none.fl_str_mv IMIQUIMOD
HEMANGIOMA
ENDOTELIO
MITOCONDRIA
topic IMIQUIMOD
HEMANGIOMA
ENDOTELIO
MITOCONDRIA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The off-label use of imiquimod (IQ) for hemangioma treatment has shown clinical benefits. We have previously reported a selective di- rect IQ-cytotoxic effect on transformed H5V endothelial cells (EC) (hemangioma model) vs normal 1G11 EC. We observed a severe imbalance in antioxidant defense and apoptosis in H5V but not in 1G11. To further address this issue, we studied the possibility of IQ being a mitochondrial toxicant. H5V and 1G11 cells were treated with IQ (0-50 μg/mL) for 2, 4, 12 or 24 h and analyzed for reac- tive oxygen species (ROS) with DCFH2-DA probe and mitochondrial stress by MitoTrackerTM Red CMXRos fluorescence. Viability assays were performed using the standard culture medium with 5.5 mM glucose (regular) or media containing 25 mM glucose (high) or 25 mM galactose (depleted). IQ treatment increased ROS level in H5V after 2 h (35-60%; p<0.05) but in 1G11 only at 4 h (̴50%; p<0.05). Mitochondrial membrane potential in H5V cells was affected after 4 and 12 h treatment, revealed by a decreased in MitoTracker fluores- cence (≈50%; p<0.05). In contrast, 1G11 cells were unaffected and only presented a significant 30%-decrease in fluorescence after 12 h with 50 μg/mL IQ (p<0.05). Cells grown in a high glucose medium can adapt to a glycolytic phenotype. By assessing the effect of IQ in this medium, both cell lines became significantly less affected than with the regular culture medium. On the contrary, by forcing cells to respiration with galactose instead of glucose-containing medium, IQ treatment enhanced cell death in both cell lines, being fully cytotoxic for H5V (p<0.05) but leaving ≈32% 1G11 cells still alive at the high- est IQ concentrations.These results provide more evidences about the higher suscepti- bility of transformed EC to IQ, where an early ROS production and mitochondrial dysfunction drove H5V cells to death. By shifting cells towards diminished respiration in absence of glucose, we proved IQ acts as a mitochondrial toxicant in both EC lines.
Fil: Rocco, Rodrigo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Wainstok, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Cochon, Adriana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina
Fil: Gazzaniga, Silvina Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
LXVI Reunión anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión anual de la Sociedad Argentina de Immunología; LIII Reunión anual de la Asociación Argentina de Farmacología Experimental y XI Reunión anual de la Asociación Argentina de Nanomedicinas
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Asociación Argentina de Farmacología Experimental
Asociación Argentina de Nanomedicinas
description The off-label use of imiquimod (IQ) for hemangioma treatment has shown clinical benefits. We have previously reported a selective di- rect IQ-cytotoxic effect on transformed H5V endothelial cells (EC) (hemangioma model) vs normal 1G11 EC. We observed a severe imbalance in antioxidant defense and apoptosis in H5V but not in 1G11. To further address this issue, we studied the possibility of IQ being a mitochondrial toxicant. H5V and 1G11 cells were treated with IQ (0-50 μg/mL) for 2, 4, 12 or 24 h and analyzed for reac- tive oxygen species (ROS) with DCFH2-DA probe and mitochondrial stress by MitoTrackerTM Red CMXRos fluorescence. Viability assays were performed using the standard culture medium with 5.5 mM glucose (regular) or media containing 25 mM glucose (high) or 25 mM galactose (depleted). IQ treatment increased ROS level in H5V after 2 h (35-60%; p<0.05) but in 1G11 only at 4 h (̴50%; p<0.05). Mitochondrial membrane potential in H5V cells was affected after 4 and 12 h treatment, revealed by a decreased in MitoTracker fluores- cence (≈50%; p<0.05). In contrast, 1G11 cells were unaffected and only presented a significant 30%-decrease in fluorescence after 12 h with 50 μg/mL IQ (p<0.05). Cells grown in a high glucose medium can adapt to a glycolytic phenotype. By assessing the effect of IQ in this medium, both cell lines became significantly less affected than with the regular culture medium. On the contrary, by forcing cells to respiration with galactose instead of glucose-containing medium, IQ treatment enhanced cell death in both cell lines, being fully cytotoxic for H5V (p<0.05) but leaving ≈32% 1G11 cells still alive at the high- est IQ concentrations.These results provide more evidences about the higher suscepti- bility of transformed EC to IQ, where an early ROS production and mitochondrial dysfunction drove H5V cells to death. By shifting cells towards diminished respiration in absence of glucose, we proved IQ acts as a mitochondrial toxicant in both EC lines.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/249759
Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod; LXVI Reunión anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión anual de la Sociedad Argentina de Immunología; LIII Reunión anual de la Asociación Argentina de Farmacología Experimental y XI Reunión anual de la Asociación Argentina de Nanomedicinas; Argentina; 2021; 105-105
CONICET Digital
CONICET
url http://hdl.handle.net/11336/249759
identifier_str_mv Replacing glucose media with galactose to evaluate mitochondrial toxicity of imiquimod; LXVI Reunión anual de la Sociedad Argentina de Investigación Clínica; LXIX Reunión anual de la Sociedad Argentina de Immunología; LIII Reunión anual de la Asociación Argentina de Farmacología Experimental y XI Reunión anual de la Asociación Argentina de Nanomedicinas; Argentina; 2021; 105-105
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://medicinabuenosaires.com/revistas/vol81-21/s3/Mv81s3.pdf
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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application/vnd.openxmlformats-officedocument.wordprocessingml.document
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dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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