| 王 颖,冯世庆,赵 鹏,焦新旭,魏志坚,韩明远.大鼠脊髓损伤后不同时期移植人脐带间充质干细胞的修复效果观察[J].中国脊柱脊髓杂志,2010,20(11):918-925. |
| 大鼠脊髓损伤后不同时期移植人脐带间充质干细胞的修复效果观察 |
| 中文关键词: 脊髓损伤 脐带间充质干细胞 细胞移植 大鼠 |
| 中文摘要: |
| 【摘要】 目的：观察大鼠脊髓损伤后不同时期局部移植人脐带间充质干细胞（human umbilical cord mesenchy?鄄mal stem cells，hUCMSCs）修复脊髓损伤的效果，探讨hUCMSCs局部移植治疗脊髓损伤（spianl cord injury， SCI）的最佳移植时机。方法：雌性Wistar大鼠80只，随机均分为A、B、C、D组，以Impactor Model-Ⅱ打击器制备脊髓胸10（T10）损伤模型。A组为单纯损伤组，B组损伤后3d移植hUCMSCs，C组损伤后1w移植hUCM?鄄SCs，D组损伤后3w移植hUCMSCs，分别于造模后24h、移植前1d及移植后8w（A组与D组损伤后相应时间点相同）每周对各组大鼠后肢功能进行行为学评分（BBB评分）；移植后8周免疫组化染色观察移植细胞的存活、分化和血管再生情况，10周时用生物素葡聚糖胺（biotin dextran amine，BDA）示踪皮质脊髓束（corticospinal tract，CST）观察轴突的再生情况。结果：SCI后24h各组大鼠BBB评分无显著性差异（P>0.05），D组在移植前和移植后1周与A组相应时间点的BBB评分无显著性差异（P>0.05），移植后2w起各时间点评分均明显高于A组相应时间点，差异有统计学意义（P<0.05）；A组在SCI后9w时到达平台期，与实验结束时相比差异无统计学意义（P>0.05）。移植各组移植后BBB评分逐渐上升，后一个时间点与同组前一时间点比较差异有显著性意义（P<0.05），移植后6w开始C组明显高于B、D组（P<0.05）。免疫组化染色A组脊髓中未见Brdu阳性细胞，损伤区血管形态欠完整，未见新生血管；损伤周围星形胶质细胞大量增生，细胞肥大、突起增多、排列不规则，形成胶质瘢痕；未见BDA标记的皮质脊髓束通过损伤区。B、C、D组脊髓中均有Brdu阳性标记的细胞存活，C、D组中细胞数量明显多于B组，但三组中均未见到移植细胞向神经元或神经胶质样细胞分化；B、D组损伤周围的星形胶质细胞较A组减少，但排列仍欠规则；C组损伤周围星形胶质细胞形态趋于正常，排列规则，且有大量星形胶质细胞通过损伤区，血管再生数目及通过损伤区的皮质脊髓束神经纤维也明显多于B、D组。结论：大鼠SCI后局部移植的hUCMSCs能长期存活，抑制胶质瘢痕形成，改善后肢运动功能；SCI后1w移植的效果优于SCI后3d和3w时移植的效果。 |
Repairing effect of human umbilical cord mesenchymal stem cells transplantation at different time points after spinal cord injury in adult rats |
| 英文关键词:Umbilical cord mesenchymal stem cells Spinal cord injury Cell transplantation Rat |
| 英文摘要: |
| 【Abstract】 Objective：To observe the therapeutic effect of the human umbilical cord mesenchymal stem cells（hUCMSCs） transplantion at different time points after spinal cord injury in Wistar rats，and to explore the optimal time point of hUCMSCs transplantation for repairing spinal cord injury.Method：Eighty female Wistar rats were randomly divided into control group（group A），hUCMSCs transplantation 3d after injury（group B），hUCMSCs transplantation 1w after injury（group C） and hUCMSCs transplantation 3w after injury（group D）.The SCI animal model were induced by Impactor Model-Ⅱ at T10.Behavioral testing was performed 24 hours after injury，1 day pre-transplantation and post-transplantation weekly for 2 months according to Basso-Beattie-Bresnahan（BBB） open-field locomotor Rating Scale.Spinal cord tissues were subjected to immunohistochemical staining to detect the vascular regeneration and survival，and differentiation and migration of hUCMSCs.Result：BBB scores were no significant difference among all four groups 24 hours after SCI（P>0.05）.Before transplantation and 1 week after transplantation，there was also no significant difference between group D and group A，but group D had significantly higher score than group A at each time points from 2 weeks after transplantation（P<0.05）.The score of group A reached to a plateau at 9 weeks after SCI，which showed no statistical significance（P>0.05） compared with that at the end of experiment.BBB scores of group B，C and D increased gradually after transplantation，with significance noted between the consecutive time points in any group（P<0.05），and the score of group C was significantly higher than that of group B and D 6 weeks after transplantation（P<0.05）.Immunohistochemistry results showed absence of Brdu positive cell，abnormality of vascular shape absence of newborn vascular in group A；proliferation of glial cells with the characteristics of hypertrphy，disorder and scar formation around lesion site was evidenced.No BDA marked corticospinal tract was seen across the lesion site.Brdu positive cells were noted in group B，C and D，with the number of cells in group C and D more than that of group B.However，no differentiation to glial cell was evidenced in 3 groups.Group B and D had less and more disordered glial cells around lesion site than group A；the glial cells aroud lession site in group C was ordered and normal，with more regenerated vascular crossing corticospinal tract than that of group B and D.Conclusion：hUCMSCs transplantation after SCI can survive at least 2 months，which can inhibit glial scars formation，improve microcirculation of lesion site，and finally promote axonal regeneration as well as improving functional recovery.One week after SCI was the optimal time point. |
| 投稿时间：2010-06-07 修订日期：2010-08-03 |
| DOI：10.3969/j.issn.1004-406X.2010.[issue].918.7 |
| 基金项目:基金项目：国家自然科学基金（30872603）、天津市卫生局科技重点基金（07KG2）、天津市卫生局科研基金（09KZ104） |
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