RESUMEN
Parkinson's disease (PD) is one of the most important neurodegenerative worldwide disorders. The potential cytoprotective effects of aqueous extract of Valeriana officinalis on rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells were demonstrated. The cytotoxicity, cell viability and analysis of cellular morphology were performed by MTT-tetrazole (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and phase contrast microscopy, respectively. Significant changes in the cellular morphology, and condensation of the cell body could be observed when cells were treated with 300 nM rotenone for 48 h. Three different concentrations of Valeriana officinalis extract were used (0.049, 0.098 and 0.195 mg/mL). These extracts brought about an increase of 7.0 +/- 1.3%, 14.5 +/- 1.3% and 14.5 +/- 3.2% in cell viability. Our results indicated that neuroprotector action of the Valeriana officinalis extract provides support for later studies as they help understanding this drug for the development of cytoprotective various therapies in PD.
Asunto(s)
Citoprotección , Enfermedad de Parkinson/tratamiento farmacológico , Extractos Vegetales/uso terapéutico , Valeriana/química , Línea Celular Tumoral , HumanosRESUMEN
Cellular and subcellular organization and distribution of actin filaments have been studied with various techniques. The use of fluorescence photo-oxidation combined with phalloidin conjugates with eosin has allowed the examination of the precise cellular and subcellular location of F-actin. Correlative fluorescence light microscopy and transmission electron microscopy studies of F-actin distribution are facilitated with this method for morphological and physiological studies. Because phalloidin-eosin is smaller than other markers, this method allows the analysis of the three-dimensional location of F-actin with high-resolution light microscopy, three-d serial sections reconstructions, and electron tomography. The combination of selective staining and three-dimensional reconstructions provide a valuable tool for revealing aspects of the synaptic morphology that are not available when conventional electron microscopy is used. By applying this selective staining technique and three-dimensional imaging, we uncovered the structural organization of actin in the postsynaptic densities in physiological and pathological conditions.
Asunto(s)
Citoesqueleto de Actina/ultraestructura , Sistema Nervioso Central/ultraestructura , Colorantes Fluorescentes/farmacología , Imagenología Tridimensional/métodos , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Animales , Sistema Nervioso Central/metabolismo , Eosina Amarillenta-(YS)/farmacología , Humanos , Microscopía Fluorescente/métodos , Modelos Moleculares , Oxidación-Reducción , Faloidina/farmacología , Fotones , Coloración y Etiquetado/métodosRESUMEN
Parkinson's disease (PD) is one of the most important neurodegenerative worldwide disorders. It is characterized by a selective and progressive degeneration of dopaminergic neurons, causing a series of symptoms which might ultimately induce programmed cell death. The potential cytoprotective effects of one of the commercial extracts of Anemopaegma mirandum (Catuaba), a Brazilian tree, on Rotenone-induced apoptosis in human neuroblastomas SH-SY5Y cells was demonstrated. The cell viability, analysis of cellular morphology, nuclei morphology and ultra structural research were done by MTT-tetrazole (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, phase contrast microscopy, stained with Hoechst 33258 and electron microscopy transmission, respectively. Three different concentrations of Catuaba extract were used (0.312, 0.625 and 1.250 mg/mL). These extracts promoted an increase of 22.3+/-3.6%, 22.0+/-2.1% and 15.8+/-0.7% on the cell viability. Notable changes in the cellular morphology, condensation of the cell body, nuclear fragmentation and condensation into discrete dense chromatin clumps were observed when the cells were treated with 300 nM Rotenone for 48 h. These effects were partially altered when the extract of A. mirandum was added to the Rotenone treatment. Ultra structural analysis by electron microscopy demonstrated that citoplasmatic membranes and mitochondria membrane were also clearly preserved in the group treated with the extract. Therefore, in this study, our findings indicated that extracts of A. mirandum have cytoprotective effects on Rotenone-induced apoptosis in human neuroblastomas SH-SY5Y cells.