Sodium formate induces autophagy and apoptosis via the JNK signaling pathway of photoreceptor cells.
Mol Med Rep
; 13(2): 1111-8, 2016 Feb.
Article
em En
| MEDLINE
| ID: mdl-26676762
ABSTRACT
Incidents associated with methanol intoxication resulting from the consumption of fake wine occur not infrequently worldwide. Certain individuals are made blind due to methanol poisoning. The present study aimed to investigate the effects of sodium formate exposure on photoreceptor cells (661W cells). The 661W cells were exposed to sodium formate for 624 h and cell viability was determined using a 3(4,5dimethylthiazol2yl)2,5diphenyl2Htetrazolium bromide (MTT) assay. Subsequently, the 661W cells were exposed to 15 or 30 mM sodium formate for 24 h. The level of apoptosis was determined using Hoechst 33342/propidium iodide staining, visualizing the cells under a fluorescence microscope, and annexin Vfluorescein isothiocyanate staining, using flow cytometric analysis. Intracellular reactive oxygen species (ROS) were measured using 2',7'dichlorofluorescein diacetate (DCFHDA) staining, followed by flow cytometric analysis. Autophagy of the 661W cells was measured by monodansylcadaverine staining. The activation of phosphorylated cJun Nterminal kinase (pJNK), Bcell lymphoma (Bcl2), Bcl2associated X protein, cleaved caspase3, cleaved caspase9 and microtubuleassociated protein 1A/1Blight chain 3 (LC3) was assessed by western blotting. The effects of ZVADfmk (a pancaspase inhibitor) and SP600125 (a JNK inhibitor) on the viability of the sodium formateinduced 661W cells were determined using an MTT assay. Sodium formate treatment induced a decrease in the viability of the 661W cells in a time and a dosedependent manner. In addition, sodium formate at concentrations of 15 or 30 mM markedly increased the level of apoptosis and the ROS levels, as measured by DCFHDA staining of the 661W cells. Additionally, 661W cells exposed to sodium formate for 24 h exhibited increased levels of pJNK, Bax, cleaved caspase3, cleaved caspase9 and LC3II (the phosphatidylethanolaminemodified form of LC3), although the level of Bcl2 was decreased. Furthermore, cell cytotoxicity and autophagy were induced upon treatment with sodium formate. ZVADfmk and SP600125 were able to effectively circumvent the effects of sodium formate on cell viability. These results suggested that the cytotoxicity induced by sodium formate induces the activation of the JNK signaling pathway, leading to caspasedependent apoptosis. Increased levels of autophagy were also observed during the process of 661W cell damage induced by sodium formate.
Texto completo:
1
Coleções:
01-internacional
Temas:
Geral
Base de dados:
MEDLINE
Assunto principal:
Células Fotorreceptoras
/
Autofagia
/
Apoptose
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Sistema de Sinalização das MAP Quinases
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Proteínas Quinases JNK Ativadas por Mitógeno
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Formiatos
Limite:
Humans
Idioma:
En
Revista:
Mol Med Rep
Ano de publicação:
2016
Tipo de documento:
Article