摘要
目的探讨不同浓度谷氨酸对海马神经元的损伤作用,流式细胞仪在建立理想的兴奋性毒性模型中的应用。方法在体外原代培养10d的海马神经元中分别加入不同浓度的谷氨酸(100、200、400、600、800μmol/L),24h后相差显微镜下观察细胞形态变化,用MTT法检测存活率和流式细胞仪检测凋亡率以评定谷氨酸对海马神经元的损伤程度。结果不同浓度的谷氨酸组与对照组的细胞存活率差异有显著性(P<0.01),并呈浓度依赖性,随着谷氨酸浓度的升高,神经元的存活率降低;谷氨酸终浓度(100、200、400μmol/L)组的细胞凋亡率与对照组比较差异有显著性(P<0.01);600和800μmol/L谷氨酸组的细胞凋亡率与对照组比较差异无显著性(P>0.05),但细胞凋亡率随着谷氨酸浓度的增加而降低。结论过高浓度的谷氨酸导致细胞急性坏死而非迟发性凋亡,运用流式细胞仪检测体外培养海马神经元凋亡率是一种特异性高的检测方法,值得推广。
Objective To study the different levels of glutamate in hippocampal neurons and the application of the flow cytometry in establishing the ideal excitotoxicity model. Methods The different levels of hippocampal neurons （100,200,400,600,800 μmol/L） were added in the primary cultured hippocampal neurons and 24 hours later, we tested the neuron survival rate by using MTT method and investigated the neuron apoptosis rate by flow eytometry to evaluate the degree of injury on hippocampal neurons. Results In neuron survival rate, there was a significant differece in glutamate groups with different levels compared with the control group and there was concentration dependent （P〈0.01 ）. Three level glutamate （ 200,200,400 μmol/L） groups were compared with the control group： the significant of difference in the neuron apoptosis rate （P〉0.01）, and the subsequent two groups（600,800 μtool/L） were compared with the control group; no significant difference was fount （P〉0.05）. But the neuron apoptosis rate decreased along with the increase of the concentration of glutamate. Conclusions The higher level of glutamate can induce acute excitotoxic necrosis and not delayed apoptosis. The method of testing the hippocampal neuron apoptosis rate by flow cytometry is a specific and propagable approach.
出处
《卒中与神经疾病》
2006年第3期138-141,共4页
Stroke and Nervous Diseases
