背景:外源性神经干细胞具有神经修复作用,可能对脑出血后的神经功能恢复起到一定的作用。目的:观察胎鼠神经干细胞的体外生长、分化及移植到脑出血大鼠后的存活、迁徙、分化情况,探讨神经干细胞对脑出血模型大鼠受损神经功能的修复作用。设计:完全随机分组设计,对照动物实验。单位:复旦大学附属华山医院神经外科材料:选用健康雄性成年SD大鼠18只为受体,体质量280~320g,由中国科学院上海实验动物中心提供。实验用鼠抗BrdU为Neomarkers产品,鼠抗胶质纤维酸性蛋白和兔抗微管相关蛋白2为Chemicon产品。方法:实验于2006-02/12在复旦大学附属上海医学院解剖组胚实验室完成。从胎龄14d的胎鼠海马中分离、培养、鉴定神经干细胞。16只受体SD大鼠被随机分为3组:对照组,PBS组和移植神经干细胞组。均通过尾状核内注射自体动脉血制作大鼠脑出血模型。移植NSC组在造模后30min在血肿腔周围四点分别移植浓度为2×1011L-1神经干细胞悬液5μL;PBS组于相同时间点在脑内相同部位注射PBS;PBS和神经干细胞的移植方法同自体血的移植方法。对照组大鼠在造模后30min只造成四点损伤,不注射任何物质。主要观察指标:在造模后立即,1,3,5,14,21,28d采用前肢评分和转身评分对大鼠神经功能进行评估。大鼠于造模后28d麻醉后取脑,并通过双标胶质纤维酸性蛋白、微管相关蛋白2、BrdU免疫组化来检测移植入脑的神经干细胞在体内的分化情况。结果:①神经功能评分:造模后5d,各组差异无显著性意义(P>0.05)。造模后14~28d,干细胞移植组较其他3组明显改善(P<0.05)。②脑组织切片双免疫组织学双标染色结果:干细胞移植组血肿周围凋亡细胞少于PBS组。受体大鼠脑组织切片显示有BrdU,微管相关蛋白2,胶质纤维酸性蛋白阳性细胞,说明神经干细胞可以在宿主脑内存活、迁徙和分化,可以分化为神经元样细胞和神经胶质样细胞。结论:神经干细胞移植可能通过分化为神经元样细胞和神经胶质细胞促进大鼠脑出血的神经功能恢复。 BACKGROUND: Exogenous neural stem cells have the function of nerve repair and may play a role in the recovery of neural function after intracerebral hemorrhage. OBJECTIVE: To observe the survival, migration and differentiation of fetal rat NSCs in vitro after their differentiation, growth, differentiation and transplantation into intracerebral hemorrhage (ICH) rats, and to investigate the repair effect of neural stem cells on damaged neurological function in ICH rats. Design: Complete randomized block design, control animal experiments. SETTING: Department of Neurosurgery, Huashan Hospital affiliated to Fudan University MATERIALS: Eighteen healthy adult male Sprague-Dawley rats were used as recipients with body weight 280-320 g and were provided by Shanghai Experimental Animal Center, Chinese Academy of Sciences. Experimental mouse anti-BrdU was Neomarkers product, mouse anti-glial fibrillary acidic protein and rabbit anti-microtubule-associated protein 2 were Chemicon products. METHODS: The experiment was performed at the Anatomy Laboratory of Shanghai Medical College affiliated to Fudan University from February to February 2006. The neural stem cells were isolated, cultured and identified from fetal rat hippocampus 14 days old. Sixteen recipients of SD rats were randomly divided into three groups: control group, PBS group and transplantation of neural stem cells group. Rat models of intracerebral hemorrhage were made by injection of autologous arterial blood through caudate nucleus. Transplanted NSC group were injected 5μL of 2 × 1011L-1 neural stem cells suspension at 4 points around the hematoma cavity at 30min after modeling respectively. The PBS group was injected with the same site of brain at the same time point. PBS and neural stem cell transplantation With autologous blood transplantation method. Rats in the control group only caused four-point injury 30 min after modeling, and no substance was injected. MAIN OUTCOME MEASURES: Neurological function of rats was assessed by using forelimb score and turn score immediately after modeling. After anesthesia, rats were sacrificed at 28 days after modeling, and the differentiation of neural stem cells transplanted into the brain was detected by double-labeled glial fibrillary acidic protein, microtubule-associated protein 2 and BrdU immunohistochemistry. Results: ①Nerve function score: 5d after modeling, there was no significant difference between each group (P> 0.05). From 14 to 28 days after model establishment, the stem cell transplantation group was significantly improved compared with the other three groups (P <0.05). ② double immunohistochemistry of brain tissue sections double staining: stem cell transplantation group less apoptotic cells around the hematoma than PBS group. Receptor rat brain sections showed BrdU, microtubule-associated protein 2, glial fibrillary acidic protein-positive cells, indicating that neural stem cells can survive in the host brain, migration and differentiation, can differentiate into neuron-like cells and glia Like cells. CONCLUSION: Neural stem cell transplantation may promote neurological recovery from intracerebral hemorrhage in rats by differentiating into neuron-like cells and glial cells.