摘要
目的探讨经颈动脉神经干细胞（NSCs）移植治疗大鼠脑出血（ICH）后遗症的适宜移植时间窗和疗效。方法48只Wistar大鼠脑出血模型，随机分为实验组和对照组。分别于脑出血后第2、7、14、21、28天经颈动脉行5-溴脱氧尿嘧啶核苷（BrdU）体外标记的神经干细胞移植。对照组大鼠，仅接受颈动脉内注入等量DMEM培养液。每周测试大鼠行为功能评分；2个月后处死所有动物，行组织化学染色，观察神经干细胞在脑内的分布、分化，评定病灶体积大小。结果与对照组比较，NSCs移植组大鼠行为功能改善明显（ANOVA，P〈0．05）；脑出血第7天移植组在移植后第3周和ICH后第9周的行为功能评分显著优于其他时间移植组（SNK，P〈0．05）。ICH后第7天和第14天移植组BrdU阳性细胞计数显著多于ICH后第2、21和第28天移植组。ICH后第2天移植的NSCs绝大部分分化为星形胶质细胞（84．5±7．6）％；而ICH后第21和28天移植的NSCs分化为神经元的比例明显增大（35．4±3．1）％、（37．2±4．1）％；然而，7～14d移植组，分化为神经元的绝对数量显著高于其他治疗组（P〈0．01）。结论经皮颈动脉注射是一种可行的、微侵袭和高效的细胞移植方法。在脑出血后7～14d期间进行神经干细胞移植，能显著提高细胞存活率和移植细胞分化为神经元，明显促进功能改善。
Objective To explore optimal time window and effect of intracarotid transplantation of neural stem cells （ NSCs ） to treat neurological sequelae after experimental intracerebral hemorrhage. Methods NSCs expanded in vitro were prelablled by BrdU （5 μmol/L） , and intracerebral hemorrhage was induced by stereotaxic, intrastriatal administration of VII collagenase/heparin in 48 male Wistar rats. Forty animals were randomly divided into 5 groups to receive cell transplantation through intracarotid injection at 2,7,14,21, or 28 day after injury respectively, and 8 animals subjected to ICH only served as controls, and all rats were killed 2 months after hemorrhage. Single and double immunofluorescent staining was used to observe distribution and differentiation of the grafted NSCs, and lesion size was also measured. Resuits A large portion of BrdU positive cells survived robustly and distributed evenly in the perihematoma areas. ICH-induced neurological deficits were significantly ameliorated in animals receiving NSCs transplantation as compared with those of the untreated controls （ ANOVA, P 〈 0.05 ）. Post hoc analysis revealed that animals receiving NSCs at 7th day after ICH had significantly lower scores at both 3rd week posttransplantation and 9th week after ICH than those treated at other time points （ P 〈 0.05 ）. Animals treated at 7th and 14th day had significantly more survival cells than those treated at other time points. Grafted cells of animals receiving NSCs injection at 2nd day postinjury differentiated to a greater extent into astrocytes （ 84.5 ± 7.6 ） %, while a larger percent of neurons [ （ 35.4 ± 3.1 ） % and （ 37.2 ± 4.1 ） %, respectively] were observed in 21 and 28-day groups. However,animals treated at 7th and 14th day gained the largest number of neurons （ P 〈 0.01 ）. Conclusion The percutaneous intracarotid injection is a minimally invasive and efficient method to deliver cells to the injured hemisphere, and transplantation within 7- 14 days of injury provided significantly greater grafting efficiency and relatively more neurons derived from grafted cells than that of other time points.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2008年第4期474-476,F0003,共4页
Chinese Journal of Experimental Surgery
基金
山东省优秀中青年科学家奖励基金资助项目（0385017）,山东省卫生系统1020人才工程项目
关键词
神经干细胞
脑出血
移植
Neural stem cells
Intracerebral hemorrhage
Transplantation
