脑源性神经营养因子预处理神经干细胞移植对急性缺血性脑卒中小鼠的效果

doi:10.3969/j.issn.1006-9771.2017.11.006

摘要/Abstract

摘要： 目的比较脑源性神经营养因子(BDNF)预处理对神经干细胞(NSCs)移植修复急性缺血性脑卒中的作用。方法出生1 d C57BL/6小鼠,分离NSCs体外培养及鉴定。10周龄健康C57BL/6小鼠150只随机分为5组,A组(n=20)为假手术组,B (n=20)、C (n=20)、D (n=45)、E (n=45)组采用光化学诱导法建立急性脑缺血模型。造模24 h后,D组行单纯NSCs移植,E组行BDNF预处理的NSCs移植,C组移植等量溶剂。移植前1 d,移植后3 d、7 d、14 d、21 d、28 d,行加速转棒测试及前肢抓力测试,移植后28 d,D、E组取5只小鼠行微管相关蛋白2 (MAP-2)、神经胶质纤维酸性蛋白(GFAP)免疫荧光染色。结果移植后3 d、7 d、14 d、21 d、28 d,加速转棒时间由长到短依次为E组、D组、B组(P<0.05);移植后14 d、21 d、28 d,前肢抓力由大到小依次为E组、D组、B组(P<0.05)。移植后28 d,D组、E组均发现Edu/GFAP双阳性细胞及Edu/MAP-2双阳性细胞。结论 NSCs移植可促进缺血性脑卒中后行为功能恢复,经BDNF预处理后有更好效果。

关键词: 缺血性脑卒中, 神经干细胞, 脑源性神经营养因子, 体外培养, 移植

Abstract: Objective To explore the effect of transplantation of neural stem cells (NSCs) pretreated with brain-derived neurotrophic factor (BDNF) on acute ischemic stroke in mice.Methods NSCs from a newborn (one day) C57BL/6 mouse were isolated and cultured in vitro. A total of 150 healthy C57BL/6 mice, ten-week-old, were randomly divided into five groups, that group A accepted sham operation, and groups B, C, D and E were subjected to focal ischemia by photothrombosis. Group D was transplanted NSCs 24 hours after ischemia, while group E transplanted NSCs pretreated with BDNF and group C accepted same volume of medium. All the groups were tested with rotarod test and grip strength one day before transplantation and three, seven, 14, 21 and 28 days after transplantation. The differentiation of NSCs in groups D and E were observed immunofluorescence staining of microtubule-associated protein-2 (MAP-2) and glial fibrillary acidic protein (GFAP).Results The time on rotarod arranged from more to less was groups E, D and B (P<0.05) three, seven, 14, 21 and 28 days after transplantation, as well as grip strength 14, 21 and 28 days after transplantation (P<0.05). The Edu/GFAP positive cells and Edu/MAP-2 positive cells were found in both groups D and E.Conclusion Transplanting NSCs can promote the behavioral recovery after ischemic stroke, and it is more effective as pretreated with BDNF.

Key words: ischemic stroke, neural stem cells, brain-derived neurotrophic factor, in vitro, transplantation

中图分类号:

R743.3

王栋, 杨文桢, 侯博儒, 康军林, 任海军. 脑源性神经营养因子预处理神经干细胞移植对急性缺血性脑卒中小鼠的效果[J]. 《中国康复理论与实践》, 2017, 23(11): 1263-1272.

WANG Dong, YANG Wen-zhen, HOU Bo-ru, KANG Jun-lin, REN Hai-jun. Effect of Neural Stem Cell Transplantation Pretreated with Brain-derived Neurotrophic Factor on Acute Ischemic Stroke in Mice[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2017, 23(11): 1263-1272.

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