摘要
目的:本实验建立谷氨酸(Glu)诱导神经细胞的损伤模型,观察Glu对神经细胞的兴奋毒性作用,探索Glu诱导神经细胞损伤模型的最佳浓度,为进一步研究Glu与神经系统疾病之间的关系奠定基础。方法:原代培养新生小鼠皮层神经细胞,鉴定成功后,采用不同浓度的Glu诱导神经细胞损伤,酶标仪测定乳酸脱氢酶(LDH)漏出率,用流式细胞仪检测细胞凋亡率和死亡率,以获得Glu诱导神经细胞损伤模型的最佳浓度。结果:成功培养新生小鼠皮层神经细胞,Glu诱导神经细胞损伤呈浓度依赖性。实验中Glu浓度=100μmol/L,细胞凋亡率(%)为44.34±6.19而细胞死亡率仅为4.6±0.90说明在Glu浓度=100μmol/L诱导神经细胞,能得到较大的凋亡率和较小的死亡率。结论:成功建立Glu诱导的神经细胞损伤模型,验证Glu=100μmol/L为诱导神经凋亡的最佳浓度。
Objective： In this study, we constructed neurons injury model that induced by glutamate, observed gluta- mate-induced excitatory toxicity of neurons, explored the optimal concentration for glutamate-induced neurons injury. The purpose of this study is to lay the foundation for further study of the relationship between glutamate and nervous system diseases. Methods： cultured neonatal mouse cortical neurons and cultured 7 days for identification and then neurons were exposed to different concentration glutamate solution to induced apoptosis. The LDH leakage rate was examined by micro- plate reader and apoptosis rate were examined by flow cytometry to obtain optimal glutamate concentration for induced neu- ron injury. Results： successful cultured neonatal mouse cortical neurons, glutamate induced neurons injury with concen- tration-independent. In the study, when the concentration of glutamate was 100 μmol/L, the cell apoptosis rate was 44.34 ±6.19% while the cell death rate was only 4.6 ±0.90% implying that we could got more higher neurons apoptosis rate and lower death rate when the concentration of glutamate was 100 μmol/L, Conclusions：glutamate-induced neurons injury model was constructed successfully, proving that 100 μmol/L was the best concentration of glutamate to induce neuron cells apoptosis.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2012年第4期363-367,共5页
Chinese Journal of Neuroanatomy
基金
贵州省科技厅攻关项目(黔科合sy[2009]3054)
贵州省优秀科技教育人才省长专项资金项目(黔省专合字[2010]86号)
贵州省科学技术基金项目(黔科合J字[2011]2281号)
