β-淀粉样蛋白致胚胎鼠大脑神经干细胞凋亡作用的研究  

Study on apoptosis by amyloid-β protein in embryo-rat cerebral neural stem cells

作　　者：苏心[1] 张文治[1] 

机构地区：[1]天津市环湖医院神经细胞研究室,300060

出　　处：《中国现代神经疾病杂志》2005年第4期248-251,共4页Chinese Journal of Contemporary Neurology and Neurosurgery

摘　　要：目的探讨β-粉样蛋白(amyloid-βprotein,Aβ)对胚胎鼠神经干细胞凋亡作用的影响。淀方法体外培养7d的胚胎鼠神经干细胞,分为正常对照组和实验组,实验组根据Aβ1～40作用终浓度的不同,又分为0.1,1.0,5.0,10.0和20.0μmol/L等5个亚组,分别培养12h、24h、48h及72h。然后经四唑盐实验筛选Aβ1～40致神经干细胞发生凋亡的最佳浓度,再与神经干细胞共同培养。通过Annexin-V磷脂酰丝氨酸荧光标记染色法观察神经干细胞早期凋亡情况;用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记技术(TUNEL)进行染色,观察神经干细胞中晚期凋亡情况。结果(1)经四唑盐筛选实验确定,以终浓度为5.0μmol/L的Aβ1～40作为β-淀粉样蛋白致神经干细胞发生凋亡的最佳诱导剂量。(2)神经干细胞与Aβ1～40(5.0μmol/L)共同培养至6h时,神经干细胞发生早期凋亡,细胞内出现大量绿色荧光标记物,而对照组则未见绿色荧光标记物。(3)神经干细胞与Aβ1～40(5.0μmol/L)共同培养24h时,神经干细胞发生中晚期凋亡,细胞内可见绿色荧光标记物。结论β-粉样蛋白可导致胚胎鼠大脑神经干细胞发生凋亡。Objective To study the apoptotic effect of amyloid-β protein （Aβ） on neural stem cells（ NSCs） in rat fetal cerebrum in vitro. Methods The NSCs of rat fetal cerebrum cultured up to 7d were divided into control and experimental groups. In experimental group 5 subgroups were divided according to the end concentration of Aβ1-40 as 0.1, 1.0, 5.0, 10.0 and 20.0μmol/L and cultured 12h, 24h, 48h and 72h respectively. The optimal concentration of Aβ1-40 for inducing appotosis of NSCs was screened by MTT test,then NSCs were co-cuhivated with Aβ1-40, at the optimal concentration. The early apoptosis of NSCs was observed by Annexin-V-Fluorescence while the late apoptosis of NSCs was observed by TUNEL fluorescence staining method. Results 1） It was demonstrated by MTT screening experiment, the optimal end concentration of Aβ1-40 for inducing NSCs apoptosis was 5 μmol/L of Aβ1-40. 2） The early apoptosis of NSCs appeared when NSCs co-cultivated with 5μmol/L Aβ1-40 up to 6h. A lot of green fluorescence markers were found in apoptotic cells by Annexin-V-fluorescence staining and it was not found in the control. 3） The advanced apoptosis of NSCs appeared when NSCs co-cultivated with Aβ1-40 （5μmol/L） up to 24h and the green markers were found in cells by TUNEL fluorescence staining. Conclusion It is demonstrated that Amyloid-β protein can induce apoptosis of NSCs in rat fetal cerebrum.

关 键 词：Β-淀粉样蛋白 胚胎鼠 大脑 神经干细胞凋亡 AΒ 脑组织 

分 类 号：R749.16[医药卫生—神经病学与精神病学]