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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/250678
Ver los metadatos del registro completo
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 Técnicas2025-09-29T09:34:52Zoai: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:34982025-09-29 09:34:52.814CONICET 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 |
_version_ |
1844613082306838528 |
score |
13.070432 |