Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels

Autores
Echevarria, Maria Sol; Tenconi, Paula Estefania; Bermúdez, Vicente; Calandria, Jorgelina; Bazan, Nicolas Guillermo; Mateos, Melina Valeria
Año de publicación
2024
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Introduction Together with inflammation, oxidative stress (OS) is involved in the pathogenesis of several retinal diseases. Canonical phos- pholipases D (PLD1 and PLD2) hydrolyze phosphatidylcholine (PC) to release choline and phosphatidic acid (PA), which can be dephosphorylated by lipid phosphate phosphatases (LPPs) to diacylglycerol (DAG). We previously demonstrated that PLD1 and 2 mediate the inflammatory response of retinal pigment epithelium (RPE) cells induced by high glucose (HG) levels. Furthermore, a significant increase in reactive oxygen species (ROS) was observed in RPE cells exposed to HG. Objectives This study explores the modulation of OS mediated by PLD inhibition in RPE cells exposed to HG. Methods RPE cells (ARPE-19 and D407) were exposed to HG (33 mM) or control conditions (NG, 5.5 mM). In order to mimic PLD/LPPs activation induced by HG, cells were treated with 100 μM dilauroyl PA (DLPA), 100 μM dioctanoyl glycerol (DOG) or with 50 μM DLPA + 50 μM DOG. PLD1, PLD2, cyclooxygenase-2 (COX-2) and NADPH oxidases (NOX), were inhibited using VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM), celecoxib (10 μM) or DPI (5 μM), respectively. Immunocytochemistry, fluorescent proves and western blots assays were performed to evaluate reactive oxygen species (ROS), mitochondrial membrane potential (MMP), glutathione (GSH) levels and Nrf-2 pathway Results HG-exposure significantly increased ROS levels and reduced MMP in both RPE lines, with respect to NG. PLD inhibitors prevented both effects in an Nrf2 and COX-2-independent manner. Also, exogenously added DLPA and DOG increased OS and reduced MMP. The NOX inhibitor DPI was able to prevent OS induced in RPE cells exposed to HG and also in cells exposed to DLPA+ DOG. Conclusion Our previous findings together with results presented herein, demonstrate that PLD1 and 2 inhibition not only prevents the in- flammatory response of RPE cells, but also decreases OS generated in RPE cells exposed to HG, possibly through a reduced NOX activity but in an Nrf-2 and COX-2 independent manner. The PLD/LPP pathway constitutes a novel pharmacological target to prevent, at the same time, OS and the inflammatory response, two hallmarks of several retinal diseases.
Fil: Echevarria, Maria Sol. 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: 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
Fil: Bermúdez, 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: Calandria, Jorgelina. Louisiana State University Health Sciences Center New Orleans. School Of Medicine;
Fil: Bazan, Nicolas Guillermo. Louisiana State University Health Sciences Center New Orleans. School Of Medicine;
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
XXVIth biennial meeting of the International Society of Eye Research
Argentina
International Society of Eye Research
Materia
PHOSPHOLIPHASE D
reactive oxygen species
mitochondrial membrane potential
Nrf-2 pathway
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:11336/250678
Acceder
id CONICETDig_db37e7445c40209b72275da62fdb4d00
oai_identifier_str oai:ri.conicet.gov.ar:11336/250678
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levelsEchevarria, Maria SolTenconi, Paula EstefaniaBermúdez, VicenteCalandria, JorgelinaBazan, Nicolas GuillermoMateos, Melina ValeriaPHOSPHOLIPHASE Dreactive oxygen speciesmitochondrial membrane potentialNrf-2 pathwayhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Introduction Together with inflammation, oxidative stress (OS) is involved in the pathogenesis of several retinal diseases. Canonical phos- pholipases D (PLD1 and PLD2) hydrolyze phosphatidylcholine (PC) to release choline and phosphatidic acid (PA), which can be dephosphorylated by lipid phosphate phosphatases (LPPs) to diacylglycerol (DAG). We previously demonstrated that PLD1 and 2 mediate the inflammatory response of retinal pigment epithelium (RPE) cells induced by high glucose (HG) levels. Furthermore, a significant increase in reactive oxygen species (ROS) was observed in RPE cells exposed to HG. Objectives This study explores the modulation of OS mediated by PLD inhibition in RPE cells exposed to HG. Methods RPE cells (ARPE-19 and D407) were exposed to HG (33 mM) or control conditions (NG, 5.5 mM). In order to mimic PLD/LPPs activation induced by HG, cells were treated with 100 μM dilauroyl PA (DLPA), 100 μM dioctanoyl glycerol (DOG) or with 50 μM DLPA + 50 μM DOG. PLD1, PLD2, cyclooxygenase-2 (COX-2) and NADPH oxidases (NOX), were inhibited using VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM), celecoxib (10 μM) or DPI (5 μM), respectively. Immunocytochemistry, fluorescent proves and western blots assays were performed to evaluate reactive oxygen species (ROS), mitochondrial membrane potential (MMP), glutathione (GSH) levels and Nrf-2 pathway Results HG-exposure significantly increased ROS levels and reduced MMP in both RPE lines, with respect to NG. PLD inhibitors prevented both effects in an Nrf2 and COX-2-independent manner. Also, exogenously added DLPA and DOG increased OS and reduced MMP. The NOX inhibitor DPI was able to prevent OS induced in RPE cells exposed to HG and also in cells exposed to DLPA+ DOG. Conclusion Our previous findings together with results presented herein, demonstrate that PLD1 and 2 inhibition not only prevents the in- flammatory response of RPE cells, but also decreases OS generated in RPE cells exposed to HG, possibly through a reduced NOX activity but in an Nrf-2 and COX-2 independent manner. The PLD/LPP pathway constitutes a novel pharmacological target to prevent, at the same time, OS and the inflammatory response, two hallmarks of several retinal diseases.Fil: Echevarria, Maria Sol. 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: 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; ArgentinaFil: Bermúdez, 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: Calandria, Jorgelina. Louisiana State University Health Sciences Center New Orleans. School Of Medicine;Fil: Bazan, Nicolas Guillermo. Louisiana State University Health Sciences Center New Orleans. School Of Medicine;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; ArgentinaXXVIth biennial meeting of the International Society of Eye ResearchArgentinaInternational Society of Eye ResearchInternational Society of Eye Research2024info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/250678Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels; XXVIth biennial meeting of the International Society of Eye Research; Argentina; 2024; 589-590CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://react-profile.org/ebook/ISER2024_AbstractBook/578/Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:02:54Zoai:ri.conicet.gov.ar:11336/250678instacron: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:34982025-10-22 11:02:54.946CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
title Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
spellingShingle Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
Echevarria, Maria Sol
PHOSPHOLIPHASE D
reactive oxygen species
mitochondrial membrane potential
Nrf-2 pathway
title_short Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
title_full Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
title_fullStr Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
title_full_unstemmed Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
title_sort Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels
dc.creator.none.fl_str_mv Echevarria, Maria Sol
Tenconi, Paula Estefania
Bermúdez, Vicente
Calandria, Jorgelina
Bazan, Nicolas Guillermo
Mateos, Melina Valeria
author Echevarria, Maria Sol
author_facet Echevarria, Maria Sol
Tenconi, Paula Estefania
Bermúdez, Vicente
Calandria, Jorgelina
Bazan, Nicolas Guillermo
Mateos, Melina Valeria
author_role author
author2 Tenconi, Paula Estefania
Bermúdez, Vicente
Calandria, Jorgelina
Bazan, Nicolas Guillermo
Mateos, Melina Valeria
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv PHOSPHOLIPHASE D
reactive oxygen species
mitochondrial membrane potential
Nrf-2 pathway
topic PHOSPHOLIPHASE D
reactive oxygen species
mitochondrial membrane potential
Nrf-2 pathway
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Introduction Together with inflammation, oxidative stress (OS) is involved in the pathogenesis of several retinal diseases. Canonical phos- pholipases D (PLD1 and PLD2) hydrolyze phosphatidylcholine (PC) to release choline and phosphatidic acid (PA), which can be dephosphorylated by lipid phosphate phosphatases (LPPs) to diacylglycerol (DAG). We previously demonstrated that PLD1 and 2 mediate the inflammatory response of retinal pigment epithelium (RPE) cells induced by high glucose (HG) levels. Furthermore, a significant increase in reactive oxygen species (ROS) was observed in RPE cells exposed to HG. Objectives This study explores the modulation of OS mediated by PLD inhibition in RPE cells exposed to HG. Methods RPE cells (ARPE-19 and D407) were exposed to HG (33 mM) or control conditions (NG, 5.5 mM). In order to mimic PLD/LPPs activation induced by HG, cells were treated with 100 μM dilauroyl PA (DLPA), 100 μM dioctanoyl glycerol (DOG) or with 50 μM DLPA + 50 μM DOG. PLD1, PLD2, cyclooxygenase-2 (COX-2) and NADPH oxidases (NOX), were inhibited using VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM), celecoxib (10 μM) or DPI (5 μM), respectively. Immunocytochemistry, fluorescent proves and western blots assays were performed to evaluate reactive oxygen species (ROS), mitochondrial membrane potential (MMP), glutathione (GSH) levels and Nrf-2 pathway Results HG-exposure significantly increased ROS levels and reduced MMP in both RPE lines, with respect to NG. PLD inhibitors prevented both effects in an Nrf2 and COX-2-independent manner. Also, exogenously added DLPA and DOG increased OS and reduced MMP. The NOX inhibitor DPI was able to prevent OS induced in RPE cells exposed to HG and also in cells exposed to DLPA+ DOG. Conclusion Our previous findings together with results presented herein, demonstrate that PLD1 and 2 inhibition not only prevents the in- flammatory response of RPE cells, but also decreases OS generated in RPE cells exposed to HG, possibly through a reduced NOX activity but in an Nrf-2 and COX-2 independent manner. The PLD/LPP pathway constitutes a novel pharmacological target to prevent, at the same time, OS and the inflammatory response, two hallmarks of several retinal diseases.
Fil: Echevarria, Maria Sol. 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: 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
Fil: Bermúdez, 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: Calandria, Jorgelina. Louisiana State University Health Sciences Center New Orleans. School Of Medicine;
Fil: Bazan, Nicolas Guillermo. Louisiana State University Health Sciences Center New Orleans. School Of Medicine;
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
XXVIth biennial meeting of the International Society of Eye Research
Argentina
International Society of Eye Research
description Introduction Together with inflammation, oxidative stress (OS) is involved in the pathogenesis of several retinal diseases. Canonical phos- pholipases D (PLD1 and PLD2) hydrolyze phosphatidylcholine (PC) to release choline and phosphatidic acid (PA), which can be dephosphorylated by lipid phosphate phosphatases (LPPs) to diacylglycerol (DAG). We previously demonstrated that PLD1 and 2 mediate the inflammatory response of retinal pigment epithelium (RPE) cells induced by high glucose (HG) levels. Furthermore, a significant increase in reactive oxygen species (ROS) was observed in RPE cells exposed to HG. Objectives This study explores the modulation of OS mediated by PLD inhibition in RPE cells exposed to HG. Methods RPE cells (ARPE-19 and D407) were exposed to HG (33 mM) or control conditions (NG, 5.5 mM). In order to mimic PLD/LPPs activation induced by HG, cells were treated with 100 μM dilauroyl PA (DLPA), 100 μM dioctanoyl glycerol (DOG) or with 50 μM DLPA + 50 μM DOG. PLD1, PLD2, cyclooxygenase-2 (COX-2) and NADPH oxidases (NOX), were inhibited using VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM), celecoxib (10 μM) or DPI (5 μM), respectively. Immunocytochemistry, fluorescent proves and western blots assays were performed to evaluate reactive oxygen species (ROS), mitochondrial membrane potential (MMP), glutathione (GSH) levels and Nrf-2 pathway Results HG-exposure significantly increased ROS levels and reduced MMP in both RPE lines, with respect to NG. PLD inhibitors prevented both effects in an Nrf2 and COX-2-independent manner. Also, exogenously added DLPA and DOG increased OS and reduced MMP. The NOX inhibitor DPI was able to prevent OS induced in RPE cells exposed to HG and also in cells exposed to DLPA+ DOG. Conclusion Our previous findings together with results presented herein, demonstrate that PLD1 and 2 inhibition not only prevents the in- flammatory response of RPE cells, but also decreases OS generated in RPE cells exposed to HG, possibly through a reduced NOX activity but in an Nrf-2 and COX-2 independent manner. The PLD/LPP pathway constitutes a novel pharmacological target to prevent, at the same time, OS and the inflammatory response, two hallmarks of several retinal diseases.
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Congreso
Book
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/11336/250678
Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels; XXVIth biennial meeting of the International Society of Eye Research; Argentina; 2024; 589-590
CONICET Digital
CONICET
url http://hdl.handle.net/11336/250678
identifier_str_mv Pharmacological inhibition of the PLD pathway prevents oxidative stress in retinal pigment epithelium cells expose to high glucose levels; XXVIth biennial meeting of the International Society of Eye Research; Argentina; 2024; 589-590
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://react-profile.org/ebook/ISER2024_AbstractBook/578/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv International Society of Eye Research
publisher.none.fl_str_mv International Society of Eye Research
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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