谷氨酸对体外培养胎鼠神经干细胞的损伤作用  

Injury of neural stem cells cultured in vitro induced by glutamate of embryonic rats

作　　者：崔俐[1] 张巨[2] 林卫红[1] 马涤辉[1] 齐亚丽[1] 付海龙[1] 孙强[1] 常明[1] 吕晓红[1] 

机构地区：[1]吉林大学第一医院神经内科,吉林长春130021 [2]吉林大学中日联谊医院手外科,吉林长春130033

出　　处：《吉林大学学报（医学版）》2009年第3期451-455,579,共6页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科研基金资助课题(200300422)

摘　　要：目的:探讨谷氨酸对体外培养胎鼠神经干细胞的损伤作用,为开发神经干细胞保护药物的研究提供理论依据。方法:取妊娠15 d的Wistar大鼠胚胎脑组织行体外细胞培养,采用免疫组化方法检测证实为神经干细胞。以正常培养的神经干细胞为对照组,谷氨酸处理组加入不同浓度的谷氨酸,MTT比色法观察神经干细胞的存活率。结果:从神经干细胞球内分化的细胞既有神经元又有神经胶质细胞,免疫组化结果显示,神经元特异性烯醇化酶(NSE)染色阳性和胶质纤维酸性蛋白(GFAP)染色阳性。谷氨酸处理组给药24 h后,与对照组比较神经干细胞存活率明显降低,50μmol.L-1谷氨酸组细胞存活率为86.45%,与对照组比较差异无显著性(P>0.05);100及1 000μmol.L-1谷氨酸组干细胞存活率(分别为71.22%和15.14%)低于对照组(P<0.05,P<0.01)。结论:谷氨酸对神经干细胞有损伤作用,且呈剂量依赖关系,随着谷氨酸剂量的增加,神经干细胞的存活率逐渐降低。Objective To study the injury of the neural stem cells cultivated in vitro induced by glutamate of embryonic rats and provide theoretical basis for development of protective drugs of neural stem cells. Methods The Wistar rat （15 d of gestation） embryonic brain tissues were cultivated in vitro and the cells were confirmed as the neural stem cells with immunohistochemical method. The neural stem cells cultured normally were used as control, the different concentrations of glutamate were added, then the survival rate of neural stem cells was observed with MTT colorimetry. Results The differentiation cells from neural stem cells had both neurons and neuroglial cells that presented neuron specific enolase （NSE） and glial fibrillary acidic protein （GFAP） dying masculine respectively. After treated with glutamate for 24 h, the survival rates of neural stem cells in glutamate groups were obviously reduced compared with control group. The survival rate in 50 μmol·L^-1 glutamate group was 86. 45%, compared with control group, there was no significant difference （P〉0.05）. The survival rates of the stem cells in 100 and 1 000μmol·L^-1 glutamate groups （71.22%, 15.14%） were significantly lower than that in control group （P〈0.05, P〈0.01）. Conclusion Glutamate has function of damaging on the neural stem cells in a dosedependent manner; when the dose of glutamate increases, the survival rate of the neural stem cells decreases gradually.

关 键 词：神经干细胞 细胞培养 谷氨酸 

分 类 号：R-332[医药卫生] R971