Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10316/108002 https://doi.org/10.1016/j.redox.2017.12.018 |
Resumo: | The extracellular pH (pHe) is a key determinant of the cellular (micro)environment and needs to be maintained within strict boundaries to allow normal cell function. Here we used HEK293 cells to study the effects of pHe acidification (24h), induced by mitochondrial inhibitors (rotenone, antimycin A) and/or extracellular HCl addition. Lowering pHe from 7.2 to 5.8 reduced cell viability by 70% and was paralleled by a decrease in cytosolic pH (pHc), hyperpolarization of the mitochondrial membrane potential (Δψ), increased levels of hydroethidine-oxidizing ROS and stimulation of protein carbonylation. Co-treatment with the antioxidant α-tocopherol, the mitochondrial permeability transition pore (mPTP) desensitizer cyclosporin A and Necrostatin-1, a combined inhibitor of Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and Indoleamine 2,3-dioxygenase (IDO), prevented acidification-induced cell death. In contrast, the caspase inhibitor zVAD.fmk and the ferroptosis inhibitor Ferrostatin-1 were ineffective. We conclude that extracellular acidification induces necroptotic cell death in HEK293 cells and that the latter involves intracellular acidification, mitochondrial functional impairment, increased ROS levels, mPTP opening and protein carbonylation. These findings suggest that acidosis of the extracellular environment (as observed in mitochondrial disorders, ischemia, acute inflammation and cancer) can induce cell death via a ROS- and mPTP opening-mediated pathogenic mechanism. |
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Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cellsAcidosisMembrane potentialMitochondriaPermeability transition poreAcidsAnimalsAntimycin ACell SurvivalCellular MicroenvironmentCyclohexylaminesHEK293 CellsHumansHydrogen-Ion ConcentrationMembrane Potential, MitochondrialMitochondria, HeartPhenylenediaminesProtein CarbonylationReactive Oxygen SpeciesReceptor-Interacting Protein Serine-Threonine KinasesRotenoneThe extracellular pH (pHe) is a key determinant of the cellular (micro)environment and needs to be maintained within strict boundaries to allow normal cell function. Here we used HEK293 cells to study the effects of pHe acidification (24h), induced by mitochondrial inhibitors (rotenone, antimycin A) and/or extracellular HCl addition. Lowering pHe from 7.2 to 5.8 reduced cell viability by 70% and was paralleled by a decrease in cytosolic pH (pHc), hyperpolarization of the mitochondrial membrane potential (Δψ), increased levels of hydroethidine-oxidizing ROS and stimulation of protein carbonylation. Co-treatment with the antioxidant α-tocopherol, the mitochondrial permeability transition pore (mPTP) desensitizer cyclosporin A and Necrostatin-1, a combined inhibitor of Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and Indoleamine 2,3-dioxygenase (IDO), prevented acidification-induced cell death. In contrast, the caspase inhibitor zVAD.fmk and the ferroptosis inhibitor Ferrostatin-1 were ineffective. We conclude that extracellular acidification induces necroptotic cell death in HEK293 cells and that the latter involves intracellular acidification, mitochondrial functional impairment, increased ROS levels, mPTP opening and protein carbonylation. These findings suggest that acidosis of the extracellular environment (as observed in mitochondrial disorders, ischemia, acute inflammation and cancer) can induce cell death via a ROS- and mPTP opening-mediated pathogenic mechanism.Elsevier2018-05info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/108002http://hdl.handle.net/10316/108002https://doi.org/10.1016/j.redox.2017.12.018eng22132317Teixeira, JoséBasit, FarhanSwarts, Herman G.Forkink, MarleenOliveira, Paulo J.Willems, Peter H. G. M.Koopman, Werner J. H.info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-08-04T09:03:20Zoai:estudogeral.uc.pt:10316/108002Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T21:24:16.462632Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells |
title |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells |
spellingShingle |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells Teixeira, José Acidosis Membrane potential Mitochondria Permeability transition pore Acids Animals Antimycin A Cell Survival Cellular Microenvironment Cyclohexylamines HEK293 Cells Humans Hydrogen-Ion Concentration Membrane Potential, Mitochondrial Mitochondria, Heart Phenylenediamines Protein Carbonylation Reactive Oxygen Species Receptor-Interacting Protein Serine-Threonine Kinases Rotenone |
title_short |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells |
title_full |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells |
title_fullStr |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells |
title_full_unstemmed |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells |
title_sort |
Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells |
author |
Teixeira, José |
author_facet |
Teixeira, José Basit, Farhan Swarts, Herman G. Forkink, Marleen Oliveira, Paulo J. Willems, Peter H. G. M. Koopman, Werner J. H. |
author_role |
author |
author2 |
Basit, Farhan Swarts, Herman G. Forkink, Marleen Oliveira, Paulo J. Willems, Peter H. G. M. Koopman, Werner J. H. |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Teixeira, José Basit, Farhan Swarts, Herman G. Forkink, Marleen Oliveira, Paulo J. Willems, Peter H. G. M. Koopman, Werner J. H. |
dc.subject.por.fl_str_mv |
Acidosis Membrane potential Mitochondria Permeability transition pore Acids Animals Antimycin A Cell Survival Cellular Microenvironment Cyclohexylamines HEK293 Cells Humans Hydrogen-Ion Concentration Membrane Potential, Mitochondrial Mitochondria, Heart Phenylenediamines Protein Carbonylation Reactive Oxygen Species Receptor-Interacting Protein Serine-Threonine Kinases Rotenone |
topic |
Acidosis Membrane potential Mitochondria Permeability transition pore Acids Animals Antimycin A Cell Survival Cellular Microenvironment Cyclohexylamines HEK293 Cells Humans Hydrogen-Ion Concentration Membrane Potential, Mitochondrial Mitochondria, Heart Phenylenediamines Protein Carbonylation Reactive Oxygen Species Receptor-Interacting Protein Serine-Threonine Kinases Rotenone |
description |
The extracellular pH (pHe) is a key determinant of the cellular (micro)environment and needs to be maintained within strict boundaries to allow normal cell function. Here we used HEK293 cells to study the effects of pHe acidification (24h), induced by mitochondrial inhibitors (rotenone, antimycin A) and/or extracellular HCl addition. Lowering pHe from 7.2 to 5.8 reduced cell viability by 70% and was paralleled by a decrease in cytosolic pH (pHc), hyperpolarization of the mitochondrial membrane potential (Δψ), increased levels of hydroethidine-oxidizing ROS and stimulation of protein carbonylation. Co-treatment with the antioxidant α-tocopherol, the mitochondrial permeability transition pore (mPTP) desensitizer cyclosporin A and Necrostatin-1, a combined inhibitor of Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and Indoleamine 2,3-dioxygenase (IDO), prevented acidification-induced cell death. In contrast, the caspase inhibitor zVAD.fmk and the ferroptosis inhibitor Ferrostatin-1 were ineffective. We conclude that extracellular acidification induces necroptotic cell death in HEK293 cells and that the latter involves intracellular acidification, mitochondrial functional impairment, increased ROS levels, mPTP opening and protein carbonylation. These findings suggest that acidosis of the extracellular environment (as observed in mitochondrial disorders, ischemia, acute inflammation and cancer) can induce cell death via a ROS- and mPTP opening-mediated pathogenic mechanism. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-05 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10316/108002 http://hdl.handle.net/10316/108002 https://doi.org/10.1016/j.redox.2017.12.018 |
url |
http://hdl.handle.net/10316/108002 https://doi.org/10.1016/j.redox.2017.12.018 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
22132317 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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