The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations

Autores
Mateos, Melina Valeria; Tenconi, Paula Estefania; Bermudez, Vicente; Oresti, Gerardo Martin; Salvador, Gabriela Alejandra; Giusto, Norma Maria
Año de publicación
2018
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Purpose: Diabetic retinopathy (DR) is one of the main causes of visual dysfunction and blindness in working-age adults in which chronic hyperglycemia, oxidative stress and inflammation are key pathogenic players. The aim of this work was to study the role of the phospholipase D (PLD) pathway in retinal pigment epithelium (RPE) cells exposed to an in vitro DR model. Methods: Human RPE cell lines (ARPE-19 and D407) were exposed to high glucose (HG) concentrations (16.5 or 33 mM) or to normal glucose concentration (NG, 5.5 mM) for 4, 24 or 72 h. Osmotic controls were performed with mannitol (Man). After experimental treatment western blot (WB), immunocytochemistry and qPCR assays were performed. Cell viability was evaluated using the MTT reduction assay and PLD activity was measured as [3H]-phosphatidylethanol ([3H]-PEth) generation from [3H]-phosphatidylcholine in the presence of 0.4% ethanol. Results: Exposure to HG increased reactive oxygen species levels and caspase-3 cleavage and reduced cell viability after 72 h of incubation. In addition, short term HG exposure (4 h) induced the activation of early events, that involve PLD and ERK1/2 signaling, nuclear factor kappa B (NF-κB) nuclear translocation and IκB phosphorylation. An increment in pro-inflammatory interleukins (IL-6 and IL-8) and cyclooxygenase-2 (COX-2) mRNA levels was observed after 24 h of HG exposure. The effect of selective pharmacological PLD1 (VU0359595) and PLD2 (VU0285655-1) inhibitors demonstrated that ERK1/2 and NF-κB activation were downstream events of both PLD isoforms. The increment in IL-6 and COX-2 mRNA levels induced by HG was reduced to control levels in cells pre-incubated with both PLD inhibitors. Furthermore, the inhibition of PLD1, PLD2, the MEK/ERK pathway (with U0126) and COX-2 (with celecoxib) prevented the loss of cell viability induced by HG. Conclusions : Our results demonstrate that HG exposure induces PLD activation in RPE cells, leading to ERK1/2 activation, IκB degradation, NF-κB nuclear translocation and expression of pro-inflammatory ILs and COX-2 and reduced cell viability. Our findings are the first evidence that classical PLDs participate in the inflammatory response of RPE cells exposed to HG and leads us to postulate these signaling enzymes as potential therapeutic targets for DR treatment
Fil: Mateos, Melina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Tenconi, Paula Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Bermudez, Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Oresti, Gerardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Salvador, Gabriela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Giusto, Norma Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
2018 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
Hawaii
Estados Unidos
Association for Research in Vision and Ophthalmology
Materia
RETINAL PIGMENT EPITHELIUM
DIABETIC RETINOPATHY
PHOSPHOLIPASE D
INFLAMMATION
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:11336/252204
Acceder
id CONICETDig_146b804d63cf2f00c7ef10707bdd02ca
oai_identifier_str oai:ri.conicet.gov.ar:11336/252204
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrationsMateos, Melina ValeriaTenconi, Paula EstefaniaBermudez, VicenteOresti, Gerardo MartinSalvador, Gabriela AlejandraGiusto, Norma MariaRETINAL PIGMENT EPITHELIUMDIABETIC RETINOPATHYPHOSPHOLIPASE DINFLAMMATIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Purpose: Diabetic retinopathy (DR) is one of the main causes of visual dysfunction and blindness in working-age adults in which chronic hyperglycemia, oxidative stress and inflammation are key pathogenic players. The aim of this work was to study the role of the phospholipase D (PLD) pathway in retinal pigment epithelium (RPE) cells exposed to an in vitro DR model. Methods: Human RPE cell lines (ARPE-19 and D407) were exposed to high glucose (HG) concentrations (16.5 or 33 mM) or to normal glucose concentration (NG, 5.5 mM) for 4, 24 or 72 h. Osmotic controls were performed with mannitol (Man). After experimental treatment western blot (WB), immunocytochemistry and qPCR assays were performed. Cell viability was evaluated using the MTT reduction assay and PLD activity was measured as [3H]-phosphatidylethanol ([3H]-PEth) generation from [3H]-phosphatidylcholine in the presence of 0.4% ethanol. Results: Exposure to HG increased reactive oxygen species levels and caspase-3 cleavage and reduced cell viability after 72 h of incubation. In addition, short term HG exposure (4 h) induced the activation of early events, that involve PLD and ERK1/2 signaling, nuclear factor kappa B (NF-κB) nuclear translocation and IκB phosphorylation. An increment in pro-inflammatory interleukins (IL-6 and IL-8) and cyclooxygenase-2 (COX-2) mRNA levels was observed after 24 h of HG exposure. The effect of selective pharmacological PLD1 (VU0359595) and PLD2 (VU0285655-1) inhibitors demonstrated that ERK1/2 and NF-κB activation were downstream events of both PLD isoforms. The increment in IL-6 and COX-2 mRNA levels induced by HG was reduced to control levels in cells pre-incubated with both PLD inhibitors. Furthermore, the inhibition of PLD1, PLD2, the MEK/ERK pathway (with U0126) and COX-2 (with celecoxib) prevented the loss of cell viability induced by HG. Conclusions : Our results demonstrate that HG exposure induces PLD activation in RPE cells, leading to ERK1/2 activation, IκB degradation, NF-κB nuclear translocation and expression of pro-inflammatory ILs and COX-2 and reduced cell viability. Our findings are the first evidence that classical PLDs participate in the inflammatory response of RPE cells exposed to HG and leads us to postulate these signaling enzymes as potential therapeutic targets for DR treatmentFil: Mateos, Melina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Tenconi, Paula Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Bermudez, Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; ArgentinaFil: Oresti, Gerardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Salvador, Gabriela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; ArgentinaFil: Giusto, Norma Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina2018 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)HawaiiEstados UnidosAssociation for Research in Vision and OphthalmologyInvestigative Ophthalmology & Visual Science2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/252204The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations; 2018 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO); Hawaii; Estados Unidos; 2018; 1-10146-04041552-5783CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://iovs.arvojournals.org/article.aspx?articleid=2692736&resultClick=1Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:37:32Zoai:ri.conicet.gov.ar:11336/252204instacron: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:34982025-09-29 09:37:32.338CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
title The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
spellingShingle The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
Mateos, Melina Valeria
RETINAL PIGMENT EPITHELIUM
DIABETIC RETINOPATHY
PHOSPHOLIPASE D
INFLAMMATION
title_short The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
title_full The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
title_fullStr The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
title_full_unstemmed The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
title_sort The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations
dc.creator.none.fl_str_mv Mateos, Melina Valeria
Tenconi, Paula Estefania
Bermudez, Vicente
Oresti, Gerardo Martin
Salvador, Gabriela Alejandra
Giusto, Norma Maria
author Mateos, Melina Valeria
author_facet Mateos, Melina Valeria
Tenconi, Paula Estefania
Bermudez, Vicente
Oresti, Gerardo Martin
Salvador, Gabriela Alejandra
Giusto, Norma Maria
author_role author
author2 Tenconi, Paula Estefania
Bermudez, Vicente
Oresti, Gerardo Martin
Salvador, Gabriela Alejandra
Giusto, Norma Maria
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv RETINAL PIGMENT EPITHELIUM
DIABETIC RETINOPATHY
PHOSPHOLIPASE D
INFLAMMATION
topic RETINAL PIGMENT EPITHELIUM
DIABETIC RETINOPATHY
PHOSPHOLIPASE D
INFLAMMATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Purpose: Diabetic retinopathy (DR) is one of the main causes of visual dysfunction and blindness in working-age adults in which chronic hyperglycemia, oxidative stress and inflammation are key pathogenic players. The aim of this work was to study the role of the phospholipase D (PLD) pathway in retinal pigment epithelium (RPE) cells exposed to an in vitro DR model. Methods: Human RPE cell lines (ARPE-19 and D407) were exposed to high glucose (HG) concentrations (16.5 or 33 mM) or to normal glucose concentration (NG, 5.5 mM) for 4, 24 or 72 h. Osmotic controls were performed with mannitol (Man). After experimental treatment western blot (WB), immunocytochemistry and qPCR assays were performed. Cell viability was evaluated using the MTT reduction assay and PLD activity was measured as [3H]-phosphatidylethanol ([3H]-PEth) generation from [3H]-phosphatidylcholine in the presence of 0.4% ethanol. Results: Exposure to HG increased reactive oxygen species levels and caspase-3 cleavage and reduced cell viability after 72 h of incubation. In addition, short term HG exposure (4 h) induced the activation of early events, that involve PLD and ERK1/2 signaling, nuclear factor kappa B (NF-κB) nuclear translocation and IκB phosphorylation. An increment in pro-inflammatory interleukins (IL-6 and IL-8) and cyclooxygenase-2 (COX-2) mRNA levels was observed after 24 h of HG exposure. The effect of selective pharmacological PLD1 (VU0359595) and PLD2 (VU0285655-1) inhibitors demonstrated that ERK1/2 and NF-κB activation were downstream events of both PLD isoforms. The increment in IL-6 and COX-2 mRNA levels induced by HG was reduced to control levels in cells pre-incubated with both PLD inhibitors. Furthermore, the inhibition of PLD1, PLD2, the MEK/ERK pathway (with U0126) and COX-2 (with celecoxib) prevented the loss of cell viability induced by HG. Conclusions : Our results demonstrate that HG exposure induces PLD activation in RPE cells, leading to ERK1/2 activation, IκB degradation, NF-κB nuclear translocation and expression of pro-inflammatory ILs and COX-2 and reduced cell viability. Our findings are the first evidence that classical PLDs participate in the inflammatory response of RPE cells exposed to HG and leads us to postulate these signaling enzymes as potential therapeutic targets for DR treatment
Fil: Mateos, Melina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Tenconi, Paula Estefania. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Bermudez, Vicente. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina
Fil: Oresti, Gerardo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Salvador, Gabriela Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
Fil: Giusto, Norma Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina
2018 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO)
Hawaii
Estados Unidos
Association for Research in Vision and Ophthalmology
description Purpose: Diabetic retinopathy (DR) is one of the main causes of visual dysfunction and blindness in working-age adults in which chronic hyperglycemia, oxidative stress and inflammation are key pathogenic players. The aim of this work was to study the role of the phospholipase D (PLD) pathway in retinal pigment epithelium (RPE) cells exposed to an in vitro DR model. Methods: Human RPE cell lines (ARPE-19 and D407) were exposed to high glucose (HG) concentrations (16.5 or 33 mM) or to normal glucose concentration (NG, 5.5 mM) for 4, 24 or 72 h. Osmotic controls were performed with mannitol (Man). After experimental treatment western blot (WB), immunocytochemistry and qPCR assays were performed. Cell viability was evaluated using the MTT reduction assay and PLD activity was measured as [3H]-phosphatidylethanol ([3H]-PEth) generation from [3H]-phosphatidylcholine in the presence of 0.4% ethanol. Results: Exposure to HG increased reactive oxygen species levels and caspase-3 cleavage and reduced cell viability after 72 h of incubation. In addition, short term HG exposure (4 h) induced the activation of early events, that involve PLD and ERK1/2 signaling, nuclear factor kappa B (NF-κB) nuclear translocation and IκB phosphorylation. An increment in pro-inflammatory interleukins (IL-6 and IL-8) and cyclooxygenase-2 (COX-2) mRNA levels was observed after 24 h of HG exposure. The effect of selective pharmacological PLD1 (VU0359595) and PLD2 (VU0285655-1) inhibitors demonstrated that ERK1/2 and NF-κB activation were downstream events of both PLD isoforms. The increment in IL-6 and COX-2 mRNA levels induced by HG was reduced to control levels in cells pre-incubated with both PLD inhibitors. Furthermore, the inhibition of PLD1, PLD2, the MEK/ERK pathway (with U0126) and COX-2 (with celecoxib) prevented the loss of cell viability induced by HG. Conclusions : Our results demonstrate that HG exposure induces PLD activation in RPE cells, leading to ERK1/2 activation, IκB degradation, NF-κB nuclear translocation and expression of pro-inflammatory ILs and COX-2 and reduced cell viability. Our findings are the first evidence that classical PLDs participate in the inflammatory response of RPE cells exposed to HG and leads us to postulate these signaling enzymes as potential therapeutic targets for DR treatment
publishDate 2018
dc.date.none.fl_str_mv 2018
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status_str publishedVersion
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dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/252204
The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations; 2018 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO); Hawaii; Estados Unidos; 2018; 1-1
0146-0404
1552-5783
CONICET Digital
CONICET
url http://hdl.handle.net/11336/252204
identifier_str_mv The phospholipase D pathway modulates the inflammatory response of retinal pigment epithelium cells exposed to high glucose concentrations; 2018 Annual Meeting of the Association for Research in Vision and Ophthalmology (ARVO); Hawaii; Estados Unidos; 2018; 1-1
0146-0404
1552-5783
CONICET Digital
CONICET
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dc.publisher.none.fl_str_mv Investigative Ophthalmology & Visual Science
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