Extracellular acidification induces ROS- and mPTP-mediated death in HEK293 cells

Detalhes bibliográficos
Autor(a) principal: Teixeira, José
Data de Publicação: 2018
Outros Autores: Basit, Farhan, Swarts, Herman G., Forkink, Marleen, Oliveira, Paulo J., Willems, Peter H. G. M., Koopman, Werner J. H.
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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spelling 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
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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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