中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (14): 2545-2550.doi: 10.3969/j.issn.1673-8225.2011.14.017

• 干细胞移植 stem cell transplantation • 上一篇    下一篇

侧脑室移植骨髓基质细胞对脑缺血再灌注大鼠运动及认知功能的影响

顾  平1，2，张忠霞1，张宝华2，王彦永2，张振清2，崔冬生1，耿  媛1，王铭维1，2   

  1. 1河北省脑老化与认知神经科学实验室，河北省石家庄市  050031
    2河北医科大学第一医院神经内科，河北省石家庄市  050031
  • 收稿日期:2010-11-05 修回日期:2010-12-31 出版日期:2011-04-02 发布日期:2013-11-02
  • 通讯作者: 王铭维，博士，主任医师，教授，博士生导师，河北省脑老化与认知神经科学实验室，河北医科大学第一医院神经内科，河北省石家庄市 050031
  • 作者简介:顾平☆，女，1970年生，江苏省无锡市人，汉族，1993年河北医科大学毕业，博士，主任医师，教授，硕士生导师，主要从事神经病学方面的研究。 gpwh2000@yahoo.com.cn
  • 基金资助:

    河北省应用基础研究计划重点基础研究项目(08966102D)。课题名称：骨髓干细胞的不同方法移植治疗大鼠缺血性脑梗死疗效的比较研究。

Effects of lateral ventricle transplantation of bone marrow stromal cells on the movement and cognitive function of cerebral ischemic reperfusion rats

Gu Ping1, 2 , Zhang Zhong-xia1, Zhang Bao-hua2, Wang Yan-yong1, 2, Zhang Zhen-qing2, Cui Dong-sheng1, Geng Yuan1, Wang Ming-wei1, 2    

  1. 1Brain Aging and Cognitive Neuroscience Laboratory of Hebei Province, Shijiazhuang  050031, Hebei Province, China
    2Department of Neurology, First Hospital of Hebei Medical University, Shijiazhuang  050031, Hebei Province, China
  • Received:2010-11-05 Revised:2010-12-31 Online:2011-04-02 Published:2013-11-02
  • Contact: Wang Ming-wei, Doctor, Chief physician, Professor, Doctoral supervisor, Brain Aging and Cognitive Neuroscience Laboratory of Hebei Province, Shijiazhuang 050031, Hebei Province, China; Department of Neurology, First Hospital of Hebei Medical University, Shijiazhuang 050031, Hebei Province, China
  • About author:Gu Ping☆, Doctor, Chief physician, Professor, Master’s supervisor, Brain Aging and Cognitive Neuroscience Laboratory of Hebei Province, Shijiazhuang 050031, Hebei Province, China; Department of Neurology, First Hospital of Hebei Medical University, Shijiazhuang 050031, Hebei Province, China
  • Supported by:

    the Key Basic Research Project of Hebei Provincial Applied Basic Research Planning, No. 08966102D*

PDF

383

被引次数

5

摘要：

背景：将骨髓基质细胞经侧脑室移植治疗大脑中动脉阻断缺血所致脑梗死模型鼠已取得一定效果，但尚未见对其认知功能的影响。
目的：观察骨髓基质细胞侧脑室移植对大脑中动脉阻断缺血模型大鼠行为认知功能的影响，并观察脑梗死灶大小的变化和植入骨髓基质细胞的迁移路径。
方法：制作SD大鼠大脑中动脉阻断缺血2 h再灌注模型后随机分为3组，骨髓基质细胞组及磷酸缓冲液组分别于梗死侧侧脑室注射骨髓基质细胞悬液5 μL(含约1.0×106个细胞)或等量的磷酸缓冲液，模型组和不造模的正常组不作任何处理。应用平衡木实验观察大鼠运动协调能力，水迷宫实验观察其游泳速度及空间学习记忆能力；苏木精-伊红染色观察梗死灶大小的变化，免疫组化观察BrdU阳性细胞的迁移路径。
结果与结论：骨髓基质细胞组在移植后第3，7，14天平衡木评分均有显著改善(P < 0.05)。移植后第7~10天，骨髓基质细胞组大鼠的游泳速度均快于磷酸缓冲液组(P  < 0.05)。骨髓基质细胞组大鼠寻找平台潜伏期的时间明显缩短(P < 0.05)，在原平台象限所占时间百分比和路程百分比及穿越平台次数明显增加(P < 0.05)。移植后7，14 d，各模型组大鼠苏木精-伊红染色均可见典型的脑缺血梗死病灶，梗死面积百分比无明显差异。BrdU染色显示，移植后第1天，阳性细胞都聚集在移植侧侧脑室，以紧密排列的细胞团形式存在于脑室壁；第3天大部分细胞穿越脑室壁以单个细胞形式向周围缺血区迁移；第14天移植细胞在梗死的纹状体、皮质可见。提示大脑中动脉阻断缺血后24 h侧脑室移植骨髓基质细胞可明显改善运动协调能力及空间学习记忆能力；移植骨髓基质细胞未能减小梗死灶大小，但侧脑室移植的骨髓基质细胞可存活并定向性地迁移至缺血的纹状体和皮质。

关键词: 骨髓基质细胞, 侧脑室, 移植, 脑缺血, 运动, 认知, 大鼠

Abstract:

BACKGROUND: Some results of improved motor function have been achieved by the transplantation of bone marrow stromal cells (BMSCs) into lateral cerebral ventricle to treat middle cerebral artery obstruction (MCAO) rats, while the cognitive function has not been reported.
OBJECTIVE: To observe the movement, cognitive function, infarction size of the MCAO rats, and the migration way of the transplanted BMSCs.    
METHODS: MCAO 2 hours reperfusion cerebral ischemia models were manufactured and were randomly divided into 3 groups: BMSCs group, phosphate buffered solution (PBS) group, and model group. The suspension of BMSCs (5 μL, 1.0x106/μL) labled with BrdU and the same volume of phosphate buffered solution were injected into the infarction side lateral cerebral ventricle of BMSCs group and PBS group respectively, while model group and normal group received no treatment. The ability of motor coordination of the MCAO rats was observed by balancing beam experiment, the swimming speed and the ability of space learning and memory were detected by morris water maze test. The infarction size was detected by hematoxylin-eosin staining, and the migration way of the BrdU positive cells was detected by immunohistochemistry staining.
RESULTS AND CONCLUSION: The score of the balance beam test in BMSCs group was significantly improved on the 3rd, 7th, 14th day after transplantation (P  < 0.05). On the 7th-10th days after transplantation, the swimming speed in the BMSCs group was significantly faster than that of PBS group (P < 0.05). The escape lantency in BMSCs group was significantly decreased (P  < 0.05), while the percentage of swimming time (T1%), swimming distance (D1%) in the plantform quadrant and the crossing times in BMSCs group were significantly increased (P < 0.05). Typical pathomorph of cerebral infarction could be seen on 7th and 14th day after transplantation in hematoxylin-eosin staining of model group rats. There was no obvious difference among the percentage of infarction area. BrdU staining showed that on the 1st day after transplantation, the positive cells were gathered in the transplanted lateral cerebral ventricle, resided in the wall of the cerebral ventricle in the style of tigh disposed cell clump. On the 3rd day, the most positive cells passed through the cerebral ventricle parietes directily migrated to the ischemic zone in the form of single cell. On the 14th day, the most positive cells were found in the striatum and cortex of the ischemia hemisphere. Results indicated that BMSCs lateral ventricle transplantation 24 hours after MCAO operation can significantly improve the motor coordination ability and the function of space learning and memory of MCAO rats. The infarction size of MCAO rats was not diminished, while the BMSCs with lateral ventricle transplantation can survive and migrate to the striatum and cortical area of cerebral ischemia zone directily.

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