Protective effects of breviscapine against cultured rat hippocampal neuronal toxicity induced by glutamate / 药学学报
Acta Pharmaceutica Sinica
; (12): 583-588, 2007.
Article
em Zh
| WPRIM
| ID: wpr-281872
Biblioteca responsável:
WPRO
ABSTRACT
The aim of this study was to investigate the effects of breviscapine on cultured rat hippocampal neuronal toxicity induced by glutamate. Primary hippocampal neurons were prepared from 2 day-old SD rats. After 8 days cultured in vitro, the cultures subjected to 30 min treatment of 0.1, 0.5 and 1.0 mmol x L(-1) L-glutamate, separately. Breviscapine (10, 20 and 40 micromol x L(-1)) was added into the cultures during 30 min treatment of L-glutamate and for the following 24 h respectively. After 24 h of L-glutamate treatment, flow cytometric analysis of Annexin V (marks apoptosis) and PI (propidium iodide, marks necrosis) labeling cells showed that L-glutamate dose-dependently induced hippocampal neuronal apoptosis and necrosis. In agreement with these results, RT-PCR experiments indicated a biphasic regulation of X-chromosome-linked inhibitor of apoptosis protein (XIAP) mRNA after L-glutamate treatment, i. e up-regulation by 0.1 mmol x L(-1) L-glutamate and down-regulation by 0.5 and 1.0 mmol x L(-1) L-glutamate. However, breviscapine markedly reduced apoptosis and necrosis due to toxicity of 0.5 mmol L(-1) L-glutamate. Compared with the vehicle-treated L-glutamate group, the apoptosis was reduced by 30.4% and 40.1%, and necrosis was reduced by 32.5% and 38.8%, after treatment by breviscapine of 20 and 40 micromol x L(-1). Meanwhile, breviscapine obviously reversed the down-regulation of XIAP expression induced by L-glutamate (up-regulation by 45.1% and 54.9% when compared with that of the vehicle-treated glutamate group). The results from the detection of confocal laser scanning microscopy with Fluo-3, a Ca2+ probe showed an obvious increase in intracellular Ca2+ during L-glutamate treatment; and breviscapine of 20 or 40 micromol x L(-1) significantly slowed down glutamate-induced Ca2+ influx and lowered the intracellular Ca2+ peak in hippocampal neurons (P < 0.01). These results suggest that neuroprotective effect of breviscapine against glutamate excitotoxicity was associated with inhibition of the accumulation of intracellular Ca2+ and up-regulation of XIAP expression in hippocampal neurons.
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Base de dados:
WPRIM
Assunto principal:
Farmacologia
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Flavonoides
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RNA Mensageiro
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Células Cultivadas
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Cálcio
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Ratos Sprague-Dawley
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Apoptose
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Fármacos Neuroprotetores
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Ácido Glutâmico
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Biologia Celular
Limite:
Animals
Idioma:
Zh
Revista:
Acta Pharmaceutica Sinica
Ano de publicação:
2007
Tipo de documento:
Article