Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels

Autores
Tenconi, Paula Estefania; Bermudez, Vicente; Echevarria, Maria Sol; Asatryan, Aram; Calandria, Jorgelina; Giusto, Norma Maria; Bazan, Nicolas G.; Mateos, Melina Valeria
Año de publicación
2022
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Objective: Inflammation and oxidative stress are a key factor in retinal and ocular diseases. We previously described the participation of classical phospholipase D isoforms (PLD1 and PLD2) in the inflammatory response of human retinal pigment epithelium (RPE) cells exposed to high glucose concentrations (HG). The aim of this work was to study the role of the PLD pathway in RPE phagocytic function. Materials and Methods: Two human RPE cell lines were used, ARPE-19 and ABC cells. ARPE-19 cells were exposed to HG (33 mM) or to normal glucose concentration (NG, 5.5 mM, control condition) for 24, 48 or 72 h and then non-specific phagocytosis was measured using pHrodo™ green bioparticles®. Also, photoreceptor outer segments (POS) phagocytosis was measured. To inhibit PLD1 and PLD2 cells were treated with 0.5 or 5 μM pharmacological PLD1 (VU0359595 or PLD1i) or PLD2 (VU0285655-1 or PLD2i) selective inhibitors. Also, PLD1 and PLD2 siRNA were used. Cell viability was measured by flow cytometry using propidium iodide (PI). Results: HG exposure for 48 and 72 h reduced phagocytic function of ARPE-19 cells. The loss of function induced by HG was prevented when cells were treated with 5 μM PLD1i or PLD2i. Furthermore, PLD1i and PLD2i did not affect RPE phagocytic function under physiological conditions and were able to prevent the increase in reactive oxygen species (ROS) induced by HG in RPE cells. In addition, RNAseq analyses showed for the first time PLD1 and PLD2 expression in hRPE49 and ABC cells, a novel human RPE cell line. Under physiological conditions, PLD1i and PLD2i did not affect ABC cells viability and partial silencing of both PLDs did not affect ABC cells POS phagocytosis. Conclusion: Our findings demonstrate that PLD1i and PLD2i prevent the loss of phagocytic function of RPE cells exposed to HG, without affecting RPE function or viability under non-inflammatory conditions, pointing to their potential use as therapeutic agents for retinal inflammatory diseases.
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: 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: 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: Asatryan, Aram. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;
Fil: Calandria, Jorgelina. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;
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
Fil: Bazan, Nicolas G.. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;
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
Reunión Conjunta SAIC SAI & FAIC SAFIS 2022; LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad Argentina de Inmunología & 3er Congreso Franco Argentino de Inmunología; Reunión Anual 2022 de la Sociedad Argentina de Fisiología
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Sociedad Argentina de Fisiología
Materia
FOSFOLIPASAS D
RETINA
FAGOCITOSIS
ALTA GLUCOSA
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:11336/228219
Acceder
id CONICETDig_6befd9f999d70dd488b66a60e318d3ce
oai_identifier_str oai:ri.conicet.gov.ar:11336/228219
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levelsTenconi, Paula EstefaniaBermudez, VicenteEchevarria, Maria SolAsatryan, AramCalandria, JorgelinaGiusto, Norma MariaBazan, Nicolas G.Mateos, Melina ValeriaFOSFOLIPASAS DRETINAFAGOCITOSISALTA GLUCOSAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Objective: Inflammation and oxidative stress are a key factor in retinal and ocular diseases. We previously described the participation of classical phospholipase D isoforms (PLD1 and PLD2) in the inflammatory response of human retinal pigment epithelium (RPE) cells exposed to high glucose concentrations (HG). The aim of this work was to study the role of the PLD pathway in RPE phagocytic function. Materials and Methods: Two human RPE cell lines were used, ARPE-19 and ABC cells. ARPE-19 cells were exposed to HG (33 mM) or to normal glucose concentration (NG, 5.5 mM, control condition) for 24, 48 or 72 h and then non-specific phagocytosis was measured using pHrodo™ green bioparticles®. Also, photoreceptor outer segments (POS) phagocytosis was measured. To inhibit PLD1 and PLD2 cells were treated with 0.5 or 5 μM pharmacological PLD1 (VU0359595 or PLD1i) or PLD2 (VU0285655-1 or PLD2i) selective inhibitors. Also, PLD1 and PLD2 siRNA were used. Cell viability was measured by flow cytometry using propidium iodide (PI). Results: HG exposure for 48 and 72 h reduced phagocytic function of ARPE-19 cells. The loss of function induced by HG was prevented when cells were treated with 5 μM PLD1i or PLD2i. Furthermore, PLD1i and PLD2i did not affect RPE phagocytic function under physiological conditions and were able to prevent the increase in reactive oxygen species (ROS) induced by HG in RPE cells. In addition, RNAseq analyses showed for the first time PLD1 and PLD2 expression in hRPE49 and ABC cells, a novel human RPE cell line. Under physiological conditions, PLD1i and PLD2i did not affect ABC cells viability and partial silencing of both PLDs did not affect ABC cells POS phagocytosis. Conclusion: Our findings demonstrate that PLD1i and PLD2i prevent the loss of phagocytic function of RPE cells exposed to HG, without affecting RPE function or viability under non-inflammatory conditions, pointing to their potential use as therapeutic agents for retinal inflammatory diseases.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; 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: 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: Asatryan, Aram. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;Fil: Calandria, Jorgelina. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;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; ArgentinaFil: Bazan, Nicolas G.. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;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; ArgentinaReunión Conjunta SAIC SAI & FAIC SAFIS 2022; LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad Argentina de Inmunología & 3er Congreso Franco Argentino de Inmunología; Reunión Anual 2022 de la Sociedad Argentina de FisiologíaMar del PlataArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de InmunologíaSociedad Argentina de FisiologíaFundación Revista Medicina2022info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/228219Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels; Reunión Conjunta SAIC SAI & FAIC SAFIS 2022; LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad Argentina de Inmunología & 3er Congreso Franco Argentino de Inmunología; Reunión Anual 2022 de la Sociedad Argentina de Fisiología; Mar del Plata; Argentina; 2022; 307-3070025-76801669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.saic.org.ar/revista-medicinaNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:03:50Zoai:ri.conicet.gov.ar:11336/228219instacron: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 10:03:50.485CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
title Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
spellingShingle Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
Tenconi, Paula Estefania
FOSFOLIPASAS D
RETINA
FAGOCITOSIS
ALTA GLUCOSA
title_short Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
title_full Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
title_fullStr Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
title_full_unstemmed Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
title_sort Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels
dc.creator.none.fl_str_mv Tenconi, Paula Estefania
Bermudez, Vicente
Echevarria, Maria Sol
Asatryan, Aram
Calandria, Jorgelina
Giusto, Norma Maria
Bazan, Nicolas G.
Mateos, Melina Valeria
author Tenconi, Paula Estefania
author_facet Tenconi, Paula Estefania
Bermudez, Vicente
Echevarria, Maria Sol
Asatryan, Aram
Calandria, Jorgelina
Giusto, Norma Maria
Bazan, Nicolas G.
Mateos, Melina Valeria
author_role author
author2 Bermudez, Vicente
Echevarria, Maria Sol
Asatryan, Aram
Calandria, Jorgelina
Giusto, Norma Maria
Bazan, Nicolas G.
Mateos, Melina Valeria
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv FOSFOLIPASAS D
RETINA
FAGOCITOSIS
ALTA GLUCOSA
topic FOSFOLIPASAS D
RETINA
FAGOCITOSIS
ALTA GLUCOSA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Objective: Inflammation and oxidative stress are a key factor in retinal and ocular diseases. We previously described the participation of classical phospholipase D isoforms (PLD1 and PLD2) in the inflammatory response of human retinal pigment epithelium (RPE) cells exposed to high glucose concentrations (HG). The aim of this work was to study the role of the PLD pathway in RPE phagocytic function. Materials and Methods: Two human RPE cell lines were used, ARPE-19 and ABC cells. ARPE-19 cells were exposed to HG (33 mM) or to normal glucose concentration (NG, 5.5 mM, control condition) for 24, 48 or 72 h and then non-specific phagocytosis was measured using pHrodo™ green bioparticles®. Also, photoreceptor outer segments (POS) phagocytosis was measured. To inhibit PLD1 and PLD2 cells were treated with 0.5 or 5 μM pharmacological PLD1 (VU0359595 or PLD1i) or PLD2 (VU0285655-1 or PLD2i) selective inhibitors. Also, PLD1 and PLD2 siRNA were used. Cell viability was measured by flow cytometry using propidium iodide (PI). Results: HG exposure for 48 and 72 h reduced phagocytic function of ARPE-19 cells. The loss of function induced by HG was prevented when cells were treated with 5 μM PLD1i or PLD2i. Furthermore, PLD1i and PLD2i did not affect RPE phagocytic function under physiological conditions and were able to prevent the increase in reactive oxygen species (ROS) induced by HG in RPE cells. In addition, RNAseq analyses showed for the first time PLD1 and PLD2 expression in hRPE49 and ABC cells, a novel human RPE cell line. Under physiological conditions, PLD1i and PLD2i did not affect ABC cells viability and partial silencing of both PLDs did not affect ABC cells POS phagocytosis. Conclusion: Our findings demonstrate that PLD1i and PLD2i prevent the loss of phagocytic function of RPE cells exposed to HG, without affecting RPE function or viability under non-inflammatory conditions, pointing to their potential use as therapeutic agents for retinal inflammatory diseases.
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: 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: 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: Asatryan, Aram. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;
Fil: Calandria, Jorgelina. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;
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
Fil: Bazan, Nicolas G.. School Of Medicine ; Louisiana State University Health Sciences Center New Orleans;
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
Reunión Conjunta SAIC SAI & FAIC SAFIS 2022; LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad Argentina de Inmunología & 3er Congreso Franco Argentino de Inmunología; Reunión Anual 2022 de la Sociedad Argentina de Fisiología
Mar del Plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Sociedad Argentina de Fisiología
description Objective: Inflammation and oxidative stress are a key factor in retinal and ocular diseases. We previously described the participation of classical phospholipase D isoforms (PLD1 and PLD2) in the inflammatory response of human retinal pigment epithelium (RPE) cells exposed to high glucose concentrations (HG). The aim of this work was to study the role of the PLD pathway in RPE phagocytic function. Materials and Methods: Two human RPE cell lines were used, ARPE-19 and ABC cells. ARPE-19 cells were exposed to HG (33 mM) or to normal glucose concentration (NG, 5.5 mM, control condition) for 24, 48 or 72 h and then non-specific phagocytosis was measured using pHrodo™ green bioparticles®. Also, photoreceptor outer segments (POS) phagocytosis was measured. To inhibit PLD1 and PLD2 cells were treated with 0.5 or 5 μM pharmacological PLD1 (VU0359595 or PLD1i) or PLD2 (VU0285655-1 or PLD2i) selective inhibitors. Also, PLD1 and PLD2 siRNA were used. Cell viability was measured by flow cytometry using propidium iodide (PI). Results: HG exposure for 48 and 72 h reduced phagocytic function of ARPE-19 cells. The loss of function induced by HG was prevented when cells were treated with 5 μM PLD1i or PLD2i. Furthermore, PLD1i and PLD2i did not affect RPE phagocytic function under physiological conditions and were able to prevent the increase in reactive oxygen species (ROS) induced by HG in RPE cells. In addition, RNAseq analyses showed for the first time PLD1 and PLD2 expression in hRPE49 and ABC cells, a novel human RPE cell line. Under physiological conditions, PLD1i and PLD2i did not affect ABC cells viability and partial silencing of both PLDs did not affect ABC cells POS phagocytosis. Conclusion: Our findings demonstrate that PLD1i and PLD2i prevent the loss of phagocytic function of RPE cells exposed to HG, without affecting RPE function or viability under non-inflammatory conditions, pointing to their potential use as therapeutic agents for retinal inflammatory diseases.
publishDate 2022
dc.date.none.fl_str_mv 2022
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
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/11336/228219
Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels; Reunión Conjunta SAIC SAI & FAIC SAFIS 2022; LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad Argentina de Inmunología & 3er Congreso Franco Argentino de Inmunología; Reunión Anual 2022 de la Sociedad Argentina de Fisiología; Mar del Plata; Argentina; 2022; 307-307
0025-7680
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/228219
identifier_str_mv Targeting phospholipase D pharmacologicallly prevents phagocytic function loss of retinal pigment epithelium cells exposed to high glucose levels; Reunión Conjunta SAIC SAI & FAIC SAFIS 2022; LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad Argentina de Inmunología & 3er Congreso Franco Argentino de Inmunología; Reunión Anual 2022 de la Sociedad Argentina de Fisiología; Mar del Plata; Argentina; 2022; 307-307
0025-7680
1669-9106
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://www.saic.org.ar/revista-medicina
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eu_rights_str_mv openAccess
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dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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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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