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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: To establish a glutamate (Glu) -induced neuronal injury model and to observe the excitotoxicity of Glu on neurons and to explore the optimal concentration of glutamate-induced neuronal cell damage model. To further investigate the relationship between Glu and neurological diseases The foundation of the relationship. Methods: Primary cultured neonatal mouse cortical neurons were cultured. After successful identification, different concentrations of Glu were used to induce neuronal injury. Lactate dehydrogenase (LDH) leakage rate was measured by microplate reader. The apoptosis rate was detected by flow cytometry And mortality in order to obtain the optimal concentration of Glu-induced neuronal cell damage model. Results: Cortical neurons were successfully cultured in neonatal rats. Glu-induced neuronal damage was dose-dependent. Glu concentration = 100μmol / L, cell apoptosis rate (%) 44.34 ± 6.19 and cell death rate 4.6 ± 0.90 in the experiment, which showed that the apoptosis rate was higher when the concentration of Glu was 100μmol / L Smaller mortality. Conclusion: The model of neuronal injury induced by Glu was successfully established. It was demonstrated that Glu = 100 μmol / L was the best concentration for inducing neuronal apoptosis.