Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 3 de 3
Filtrar
Mais filtros











Base de dados
Intervalo de ano de publicação
1.
Graefes Arch Clin Exp Ophthalmol ; 258(3): 557-564, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31938854

RESUMO

PURPOSE: To evaluate the effect of autophagy inducers on damage caused by vital dye in adult human RPE (ARPE) cells and in a rat model. METHODS: ARPE-19 cells were exposed to ICG or BBG (0.05 mg/ml) with rapamycin (200 nM) or metformin (2 mM) for 30 min and treated with or without 20 µM chloroquine (CQ) to identify the protein levels of LC3 and SQSTM1 by immunoblotting. In vivo study was performed by injecting 10 µl 0.05% ICG and 0.25% BBG into the subretinal space of the rat eyes, and/or co-treated them with metformin and rapamycin. The retinas were used to determine autophagy with the LC3-II level and apoptosis with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) assay. RESULTS: In this study, both ICG and BBG inhibited autophagy flux in adult human retinal pigment epithelium cells (ARPE-19), whereas only ICG consistently reduced autophagy in the retina of rats. Moreover, rapamycin and metformin induced autophagic flux in ARPE-19 cells and increased the LC3-II level in retinal tissues exposed to vital dyes. Both ICG and BBG increased apoptosis in the retina of rats. However, both rapamycin and metformin induced autophagy and reduced the apoptosis caused by vital dyes. CONCLUSION: Taken together, these results suggest that rapamycin and metformin may diminish vital dye-induced retinal damage in vivo through activation of autophagy.


Assuntos
Apoptose/efeitos dos fármacos , Metformina/farmacologia , Doenças Retinianas/diagnóstico , Epitélio Pigmentado da Retina/patologia , Sirolimo/farmacologia , Adulto , Animais , Sobrevivência Celular , Células Cultivadas , Corantes/toxicidade , Modelos Animais de Doenças , Humanos , Hipoglicemiantes/farmacologia , Imunossupressores/farmacologia , Ratos , Doenças Retinianas/induzido quimicamente , Doenças Retinianas/tratamento farmacológico , Epitélio Pigmentado da Retina/efeitos dos fármacos
2.
PLoS One ; 14(3): e0213932, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30870514

RESUMO

Age-related macular degeneration (AMD) is an ocular disease with retinal degeneration. Retinal pigment epithelium (RPE) degeneration is mainly caused by long-term oxidative stress. Kinase activity could be either protective or detrimental to cells during oxidative stress; however, few reports have described the role of kinases in oxidative stress. In this study, high-throughput screening of kinome siRNA library revealed that erb-b2 receptor tyrosine-protein kinase 2 (ERBB2) knockdown reduced reactive oxygen species (ROS) production in ARPE-19 cells during oxidative stress. Silencing ERBB2 caused an elevation in microtubule associated protein light chain C3-II (MAP1LC3B-II/I) conversion and sequesterone (SQSTM)1 protein level. ERBB2 deprivation largely caused an increase in autophagy-regulating protease (ATG4B) expression, a protease that negatively recycles MAP1LC3-II at the fusion step between the autophagosome and lysosome, suggesting ERBB2 might modulate ATG4B for autophagy induction in oxidative stress-stimulated ARPE-19 cells. ERBB2 knockdown also caused an accumulation of nuclear factor erythroid 2-related factor 2 (NRF2) and enhanced its transcriptional activity. In addition, ERBB2 ablation or treatment with autophagy inhibitors reduced oxidative-induced cytotoxic effects in ARPE-19 cells. Furthermore, ERBB2 silencing had little or no additive effects in ATG5/7-deficient cells. Taken together, our results suggest that ERBB2 may play an important role in modulating autophagic RPE cell death during oxidative stress, and ERBB2 may be a potential target in AMD prevention.


Assuntos
Morte Celular Autofágica/fisiologia , Proteínas Relacionadas à Autofagia/metabolismo , Cisteína Endopeptidases/metabolismo , Receptor ErbB-2/metabolismo , Epitélio Pigmentado da Retina/citologia , Epitélio Pigmentado da Retina/metabolismo , Linhagem Celular , Técnicas de Silenciamento de Genes , Humanos , Degeneração Macular/metabolismo , Degeneração Macular/patologia , Modelos Biológicos , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , RNA Interferente Pequeno/genética , Espécies Reativas de Oxigênio/metabolismo , Receptor ErbB-2/antagonistas & inibidores , Receptor ErbB-2/genética
3.
PLoS One ; 12(3): e0174736, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28358857

RESUMO

Indocyanine green (ICG) and brilliant blue G (BBG) are commonly used vital dyes to remove internal limiting membrane (ILM) in vitreoretinal surgery. The vital dyes have shown cytotoxic effects in ocular cells. Autophagy is a stress responsive pathway for either protecting cells or promoting cell death. However, the role of autophagy in ocular cells in response to the vital dyes remains unknown. In this study, we found that ICG and BBG reduced cell viability in both human retinal pigment epithelial ARPE-19 and mouse photoreceptor 661W cells. ICG and BBG induced lipidated GFP-LC3-II and LC3-II in ARPE-19 and 661W cells. Combination treatment with the autophagy inhibitor chloroquine indicated that ICG and BBG reduced autophagic flux in ARPE-19 cells, whereas the vital dyes induced autophagic flux in 661W cells. Moreover, genetic and pharmacological ablation of autophagy enhanced vital dyes-induced cytotoxicity in ocular cells. Dietary supplements, including resveratrol, lutein, and CoQ10, induced autophagy and diminished the cytotoxic effects of ICG and BBG in ocular cells. These results suggest that autophagy may protect ARPE-19 and 661W cells from vital dyes-induced damage.


Assuntos
Autofagia/efeitos dos fármacos , Corantes/efeitos adversos , Verde de Indocianina/efeitos adversos , Epitélio Pigmentado da Retina/efeitos dos fármacos , Corantes de Rosanilina/efeitos adversos , Animais , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cloroquina , Humanos , Luteína/administração & dosagem , Camundongos , Substâncias Protetoras/administração & dosagem , Resveratrol , Epitélio Pigmentado da Retina/fisiopatologia , Epitélio Pigmentado da Retina/cirurgia , Estilbenos/administração & dosagem , Ubiquinona/administração & dosagem , Ubiquinona/análogos & derivados , Vitrectomia/efeitos adversos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA