Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels

Autores
Echevarria, Maria Sol; Tenconi, Paula Estefania; Mateos, Melina Valeria
Año de publicación
2022
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Objective: Oxidative stress (OE) and inflammation are involved in the pathogenesis of several retinal diseases. We previously demonstrated that classical phospholipase D isoforms (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. The aim of the present work was to study the relationship between OE and PLD activation observed in HG- treated RPE cells. Methods: RPE cells (ARPE-19) were exposed to HG (33 mM) or to normal glucose levels (NG, 5.5 mM) for 24 h. To inhibit PLD1, PLD2 and cyclooxygenase-2 (COX-2) VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM) or celecoxib (10 μM) were used, respectively. ROS production was assessed using the probe DCDCDHF. Immunocytochemistry assays (ICC) and western blots were performed to evaluate nuclear factor erythroid 2–related factor2 (Nrf-2) pathway. Results: HG-exposure for 24 h increased ROS levels (148%) in ARPE-19 cells with respect to NG. The incubation with PLD1i and PLD2i prevented HG-induced ROS generation in RPE cells. On the contrary, the inhibition of COX-2 was not able to prevent OE induced by HG. ICC showed Nrf-2 nuclear translocation in cells exposed to HG and this effect was not observed when cells were pre-treated with PLD1i and PLD2i. Nrf-2 activation correlated with and increased heme oxygenase-1 (HO-1) expression (42%) in HG-exposed cells but no differences were observed in cells treated with PLD1i or PLD2i with respect to NG. Conclusions: Our results demonstrate that PLD1 and PLD2 inhibition not only prevents the inflammatory response of RPE cells, but also decreases OE generated in RPE cells exposed to HG in a Nrf-2 and COX-2 independent manner. Further experiments are needed to fully elucidate the mechanisms by which the PLD pathway mediates OE in RPE cells exposed to inflammatory injury.
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. 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: 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 Anual de Sociedades de Biociencias 2022: LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad argentina de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología
Mar de plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Sociedad Argentina de Fisiología
Materia
PHOSPHOLIPASE D
RETINAL PIGMENT EPITHELIUM
OXIDATIVE STRESS
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:11336/228497

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network_name_str CONICET Digital (CONICET)
spelling Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levelsEchevarria, Maria SolTenconi, Paula EstefaniaMateos, Melina ValeriaPHOSPHOLIPASE DRETINAL PIGMENT EPITHELIUMOXIDATIVE STRESShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Objective: Oxidative stress (OE) and inflammation are involved in the pathogenesis of several retinal diseases. We previously demonstrated that classical phospholipase D isoforms (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. The aim of the present work was to study the relationship between OE and PLD activation observed in HG- treated RPE cells. Methods: RPE cells (ARPE-19) were exposed to HG (33 mM) or to normal glucose levels (NG, 5.5 mM) for 24 h. To inhibit PLD1, PLD2 and cyclooxygenase-2 (COX-2) VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM) or celecoxib (10 μM) were used, respectively. ROS production was assessed using the probe DCDCDHF. Immunocytochemistry assays (ICC) and western blots were performed to evaluate nuclear factor erythroid 2–related factor2 (Nrf-2) pathway. Results: HG-exposure for 24 h increased ROS levels (148%) in ARPE-19 cells with respect to NG. The incubation with PLD1i and PLD2i prevented HG-induced ROS generation in RPE cells. On the contrary, the inhibition of COX-2 was not able to prevent OE induced by HG. ICC showed Nrf-2 nuclear translocation in cells exposed to HG and this effect was not observed when cells were pre-treated with PLD1i and PLD2i. Nrf-2 activation correlated with and increased heme oxygenase-1 (HO-1) expression (42%) in HG-exposed cells but no differences were observed in cells treated with PLD1i or PLD2i with respect to NG. Conclusions: Our results demonstrate that PLD1 and PLD2 inhibition not only prevents the inflammatory response of RPE cells, but also decreases OE generated in RPE cells exposed to HG in a Nrf-2 and COX-2 independent manner. Further experiments are needed to fully elucidate the mechanisms by which the PLD pathway mediates OE in RPE cells exposed to inflammatory injury.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. 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: 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 Anual de Sociedades de Biociencias 2022: LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad argentina de inmunología; Reunión anual 2022 de la Sociedad Argentina de FisiologíaMar de plataArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de InmunologíaSociedad Argentina de FisiologíaFundación Revista MedicinaAlonso, Daniel FernandoMalchiodi, Emilio LuisVila Petroff, Martin GerardeLamb, Caroline Ana2022info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/228497Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels; Reunión Anual de Sociedades de Biociencias 2022: LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad argentina de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología; Mar de 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 Técnicas2025-09-29T10:21:08Zoai:ri.conicet.gov.ar:11336/228497instacron: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 10:21:09.211CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
title Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
spellingShingle Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
Echevarria, Maria Sol
PHOSPHOLIPASE D
RETINAL PIGMENT EPITHELIUM
OXIDATIVE STRESS
title_short Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
title_full Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
title_fullStr Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
title_full_unstemmed Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
title_sort Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels
dc.creator.none.fl_str_mv Echevarria, Maria Sol
Tenconi, Paula Estefania
Mateos, Melina Valeria
author Echevarria, Maria Sol
author_facet Echevarria, Maria Sol
Tenconi, Paula Estefania
Mateos, Melina Valeria
author_role author
author2 Tenconi, Paula Estefania
Mateos, Melina Valeria
author2_role author
author
dc.contributor.none.fl_str_mv Alonso, Daniel Fernando
Malchiodi, Emilio Luis
Vila Petroff, Martin Gerarde
Lamb, Caroline Ana
dc.subject.none.fl_str_mv PHOSPHOLIPASE D
RETINAL PIGMENT EPITHELIUM
OXIDATIVE STRESS
topic PHOSPHOLIPASE D
RETINAL PIGMENT EPITHELIUM
OXIDATIVE STRESS
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: Oxidative stress (OE) and inflammation are involved in the pathogenesis of several retinal diseases. We previously demonstrated that classical phospholipase D isoforms (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. The aim of the present work was to study the relationship between OE and PLD activation observed in HG- treated RPE cells. Methods: RPE cells (ARPE-19) were exposed to HG (33 mM) or to normal glucose levels (NG, 5.5 mM) for 24 h. To inhibit PLD1, PLD2 and cyclooxygenase-2 (COX-2) VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM) or celecoxib (10 μM) were used, respectively. ROS production was assessed using the probe DCDCDHF. Immunocytochemistry assays (ICC) and western blots were performed to evaluate nuclear factor erythroid 2–related factor2 (Nrf-2) pathway. Results: HG-exposure for 24 h increased ROS levels (148%) in ARPE-19 cells with respect to NG. The incubation with PLD1i and PLD2i prevented HG-induced ROS generation in RPE cells. On the contrary, the inhibition of COX-2 was not able to prevent OE induced by HG. ICC showed Nrf-2 nuclear translocation in cells exposed to HG and this effect was not observed when cells were pre-treated with PLD1i and PLD2i. Nrf-2 activation correlated with and increased heme oxygenase-1 (HO-1) expression (42%) in HG-exposed cells but no differences were observed in cells treated with PLD1i or PLD2i with respect to NG. Conclusions: Our results demonstrate that PLD1 and PLD2 inhibition not only prevents the inflammatory response of RPE cells, but also decreases OE generated in RPE cells exposed to HG in a Nrf-2 and COX-2 independent manner. Further experiments are needed to fully elucidate the mechanisms by which the PLD pathway mediates OE in RPE cells exposed to inflammatory injury.
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. 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: 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 Anual de Sociedades de Biociencias 2022: LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad argentina de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología
Mar de plata
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Inmunología
Sociedad Argentina de Fisiología
description Objective: Oxidative stress (OE) and inflammation are involved in the pathogenesis of several retinal diseases. We previously demonstrated that classical phospholipase D isoforms (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. The aim of the present work was to study the relationship between OE and PLD activation observed in HG- treated RPE cells. Methods: RPE cells (ARPE-19) were exposed to HG (33 mM) or to normal glucose levels (NG, 5.5 mM) for 24 h. To inhibit PLD1, PLD2 and cyclooxygenase-2 (COX-2) VU0359595 (PLD1i, 0.5 μM), VU0285655-1 (PLD2i, 0.5 μM) or celecoxib (10 μM) were used, respectively. ROS production was assessed using the probe DCDCDHF. Immunocytochemistry assays (ICC) and western blots were performed to evaluate nuclear factor erythroid 2–related factor2 (Nrf-2) pathway. Results: HG-exposure for 24 h increased ROS levels (148%) in ARPE-19 cells with respect to NG. The incubation with PLD1i and PLD2i prevented HG-induced ROS generation in RPE cells. On the contrary, the inhibition of COX-2 was not able to prevent OE induced by HG. ICC showed Nrf-2 nuclear translocation in cells exposed to HG and this effect was not observed when cells were pre-treated with PLD1i and PLD2i. Nrf-2 activation correlated with and increased heme oxygenase-1 (HO-1) expression (42%) in HG-exposed cells but no differences were observed in cells treated with PLD1i or PLD2i with respect to NG. Conclusions: Our results demonstrate that PLD1 and PLD2 inhibition not only prevents the inflammatory response of RPE cells, but also decreases OE generated in RPE cells exposed to HG in a Nrf-2 and COX-2 independent manner. Further experiments are needed to fully elucidate the mechanisms by which the PLD pathway mediates OE in RPE cells exposed to inflammatory injury.
publishDate 2022
dc.date.none.fl_str_mv 2022
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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/11336/228497
Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels; Reunión Anual de Sociedades de Biociencias 2022: LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad argentina de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología; Mar de plata; Argentina; 2022; 307-307
0025-7680
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/228497
identifier_str_mv Phospholipase D 1 and 2 inhibition prevents oxidative stress in retinal pigment epithelium cells exposed to high glucose levels; Reunión Anual de Sociedades de Biociencias 2022: LXVII Reunión Anual de la Sociedad Argentina de Investigación Clínica; LXX Reunión Anual de la Sociedad argentina de inmunología; Reunión anual 2022 de la Sociedad Argentina de Fisiología; Mar de plata; Argentina; 2022; 307-307
0025-7680
1669-9106
CONICET Digital
CONICET
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language eng
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publisher.none.fl_str_mv Fundación Revista Medicina
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