中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (14): 2549-2555.doi: 10.3969/j.issn.1673-8225.2010.14.016

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

骨髓间充质干细胞立体定向移植治疗大鼠脊髓损伤

管雅琳1，孔繁明2，王世民3，吴  盛3，汪皖君3，唐  帆2，张文治4   

  1. 1天津医科大学研究生院，天津市   300070；天津市环湖医院，2病理科，3神经内科，  4神经外科研究所细胞室，天津市   300060
  • 出版日期:2010-04-02 发布日期:2010-04-02
  • 通讯作者: 王世民，教授，硕士生导师，天津市环湖医院神经内科，天津市 300060 wangxinpingtj@sina.com
  • 作者简介:管雅琳★，女，1982年生，山东省诸城市人，汉族，天津医科大学在读硕士，主要从事干细胞方面的研究。 Yalin82@qq.com
  • 基金资助:

    天津市卫生局科技基金支持项目(05KY17),课题名称：骨髓间充质干细胞移植治疗大鼠脊髓损伤的实验研究。

Stereotaxic transplantation of bone marrow mesenchymal stem cells in treatment of spinal cord injury in rats

Guan Ya-lin1, Kong Fan-ming2, Wang Shi-min3, Wu Sheng3, Wang Wan-jun3, Tang Fan2, Zhang Wen-zhi4   

  1. 1 Postgraduate College, Tianjin Medical University, Tianjin  300070, China; 2 Department of Pathology, 3 Department of Neurology, 4 Cell Room of Institute of Neurosurgery, Huanhu Hospital, Tianjin   300060, China
  • Online:2010-04-02 Published:2010-04-02
  • Contact: Wang Shi-min, Professor, Master’s supervisor, Department of Neurology, Huanhu Hospital, Tianjin 300060, China wangxinpingtj@sina.com
  • About author:Guan Ya-lin★, Studying for master’s degree, Postgraduate College, Tianjin Medical University, Tianjin 300070, China Yalin82@qq.com
  • Supported by:

    the Science and Technology Foundation Supporting Program of Health Bureau of Tianjin City, No. 05KY17*

PDF

591

被引次数

20

摘要：

背景：干细胞治疗神经组织损伤的关键在于移植具有再生能力的干细胞，通过多种作用机制，重建中枢神经系统的结构和功能。
目的：立体定向移植骨髓间充质干细胞观察其对大鼠脊髓损伤修复的影响并探讨其机制。
方法：密度梯度离心法结合贴壁筛选法分离骨髓间充质干细胞，于细胞移植前掺入10 mg/L的BrdU进行标记。将成年雌性Wistar大鼠以动脉瘤夹闭法制备大鼠脊髓损伤模型后随机分为对照组、生理盐水组与移植组。移植组大鼠致伤后第7天，通过立体定向途径移植骨髓间充质干细胞到脊髓损伤中心，生理盐水组给予等量生理盐水，对照组大鼠不做处理。于大鼠脊髓损伤前及损伤后第7，14，30，60，90天进行BBB评分；损伤后第90天处死大鼠，观察其脊髓组织中有无BrdU阳性细胞、Brdu+神经元特异性烯醇化酶、Brdu+胶质纤维酸性蛋白、Brdu+碱性成纤维细胞生长因子、Brdu+脑源性神经生长因子免疫组织化学双染阳性细胞及单染阳性细胞。
结果与结论： ①骨髓间充质干细胞移植组大鼠BBB后肢功能评分恢复优于对照组(P < 0.05)；生理盐水组BBB评分在损伤后30 d内恢复速度慢于对照组(P < 0.05)，至第90天与对照组比较差异无显著性意义(P > 0.05)。②移植组大鼠脊髓内在损伤中心及头、尾端距离脊髓损伤中心1 cm处均可见BrdU染色阳性细胞及免疫组织化学双染阳性细胞。移植组神经元特异性烯醇化酶、胶质纤维酸性蛋白、碱性成纤维细胞生长因子、脑源性神经生长因子单染阳性细胞数明显高于对照组和生理盐水组(P < 0.05)。结果提示，骨髓间充质干细胞移植可以促进脊髓损伤大鼠的神经功能的恢复，其机制可能与移植细胞分化为神经元样和神经胶质细胞样细胞，并分泌或促进宿主分泌神经营养因子有关。

关键词: 脊髓损伤, 骨髓间充质干细胞, 立体定向, 细胞移植, 大鼠

Abstract:

BACKGROUND: The key of stem cells for treating nervous tissue injury is the transplantation of stem cells that have regeneration capacity. The structure and function of central nervous system were re-established by multiple action mechanisms.
OBJECTIVE: To explore the effects and mechanisms of bone marrow mesenchymal stem cells (BMSCs) locally transplanted into rats with spinal cord injury on neurological recovery.
METHODS: BMSCs were separated with density gradient centrifugation and cell attachment. 10 mg/L BrdU was used for labeling before cell transplantation. Adult female Wistar rats were used to establish spinal cord injury models using an aneurysm clip, and they were then randomly divided into control group, saline group and transplantation group. In the transplantation group, BMSCs were transplanted into the damaged spinal cord by stereotaxis at day 7 following damage. In the saline group, an equal volume of saline was utilized. In the control group, the rats were left intact. Basso, Beattie and Bresnahan (BBB) locomotor rating scale was used before and at 7, 14, 30, 60 and 90 days following damage. Rats were sacrificed at day 90. BrdU-positive cells, Brdu+neuron specific enolase, Brdu+glial fibrillary acidic protein (GFAP), Brdu+basic fibroblast growth factor (bFGF), and Brdu+brain-derived nerve growth factor (BDNF) immunohistochemistry double-staining cells and simple staining positive cells were observed.
RESULTS AND CONCLUSION: The recovery of BBB function score was better in the transplantation group than in the control group (P < 0.05). The recovery speed of BBB function score was slower in the saline group than in the control group at 30 days following damage (P < 0.05). No significant difference was determined at day 90 compared with the control group (P > 0.05). BrdU-positive cells and double-staining cells of immunohistochemistry could be found at the center of damage site and 1 cm from caudal end to damaged site in rats of the transplantation group. The number of NSE, GFAP, bFGF and BDNF simple staining cells was significantly greater in the transplantation group than in the control and saline groups (P < 0.05). Results indicated that BMSC transplantation can improve the recovery of nervous function of rats with spinal cord injury. Its mechanism may be correlated with the differentiation of transplanted cells into neuron-like and glial cell-like cells, secretion or promoting secretion of neurotrophic factors in host.

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