摘要
目的探讨脑源性神经营养因子(brain derived neurotrophic factor,BDNF)对缺氧培养条件下神经干细胞(neural stem cells,NSCs)的增殖和分化的影响。方法利用MTT实验确定NSCs缺氧时间,时间范围为6、12、24、36和48 h;实验分为4组:对照组、损伤组、BDNF保护组、BDNF预保护组。对照组正常培养,其他组缺氧培养,测定各组MTT值(n=8)和乳酸脱氢酶含量(n=4);各组继续分化72 h后进行神经微管相关蛋白单抗、胶质纤维酸性蛋白多抗免疫荧光染色(n=4),计算分化率。结果由MTT法确定NSCs缺氧时间为36 h;BDNF保护组和BDNF预保护组MTT值和乳酸脱氢酶含量与损伤组比较,差异有显著性意义(P<0．01);BDNF的加入可提高NSCs分化为神经元的比率。结论BDNF对缺氧NSCs有保护作用。
Objective To study the effect of BDNF on the proliferation and differentiation of hypoxic cultured NSCs. Methods MTT method was used to determine the time of hypoxia of NSCs cultured under hypoxic condition from 6 h to 48 h. The NSCs were divided into 4 groups： the control group, the injured group, the protective group with BDNF, the pre-protective group with BDNF. The NSCs of control group were cultured under normal condition and other groups under hypoxic condition, then the MTT value and LDH content were measured after NSCs differentiated for 72 h in every group. Immunohistochemical analysis was used to detect the expression of microtubule-associated protein-2（MAP-2） and glial fibrillary acidic protein（GFAP）. The differentiation ratio was calculated. Results The hypoxic culture time of NSCs was 36 h measured by MTT method. There were significant differences in the MTT value and LDH content of the BDNF protective group and the BDNF pre-protective group as compared with the injured group（ P 〈 0.01 ）. The ratio of differentiated neural stem cells in the BDNF group was higher than that in other groups. Conclusion BDNF had protective effect on hypoxic cultured NSCs.
出处
《中华老年心脑血管病杂志》
CAS
北大核心
2007年第3期199-202,共4页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
关键词
脑源性神经营养因子
缺氧
神经干细胞
细胞保护
brain-derived neurotrophic factor
anoxia
neural stem cell
cytoprotection
