骨髓间充质干细胞动脉移植对缺血性脑损伤小鼠行为学的影响

Effect of bone-marrow mesenchymal stem cell transplantation on behavioral changes in mice after ischemic brain injury

目的研究骨髓间充质干细胞动脉移植对缺血性脑损伤小鼠行为学的影响。方法采用大脑中动脉栓塞（MCAO）模型，60只C57BL／6小鼠随机分成假手术组（sham组）、创伤组（MCAO组）和干细胞治疗组（BMSC组），后两组在造模手术拔除线栓后，由颈总动脉注入200μl PBS或间充质干细胞混悬液，假手术组只分离颈总动脉。采用TFC染色计算缺血梗死体积大小。采用神经功能局部缺损评分量表、Morris水迷宫实验、旋转平衡杆实验及加速转轮实验等评价各组小鼠的行为学改变。结果脑缺血损伤后2d，TTC染色显示BMSC组梗死体积[（34．98±12．49）％]较MCAO组缺血梗死体积小[（42．36±9．41）％]，差异有统计学意义（P〈0．05）。BMSC组与MCAO组比较，神经功能局部缺损评分在脑缺血损伤后3d起有明显改善，损伤后5d差异最显著（P〈0．01）；Morris水迷宫测试中逃避潜伏期在损伤后7d、14d显著缩短（P〈0．05），目标象限时间、游程百分比及穿越站平台次数脑损伤后均增加，脑损伤后14d差异显著（P〈0．05）；Rotarod转轮上的运动时间在脑损伤后各时间点均增高，其中脑损伤后14d差异最显著（P〈0．01）；旋转平衡杆实验中，小鼠在平衡杆上的行走速度及距离均增高，并分别于脑损伤后14d以及脑损伤后7d、10d差异最显著（P〈0．01）。结论间充质干细胞颈总动脉移植明显改善小鼠缺血脑损伤后的神经功能，促进了学习、平衡及运动功能的恢复，为临床治疗脑损伤提供了新的思路。

Objective To study the effect of bone-marrow mesenchymal stem cell （BMSC） via artery transplantation on behavior changes after ischemic brain injury in mice. Methods 60 mice （C57BL/6） were divided randomly into sham group, brain isehemia group （MCAO group） and stem cell therapy group （BMSC group）. The latter two groups were injected respectively with 200 pJ PBS or BMSC suspension into common carotid arteries namely when removal suture after middle cerebral artery occlusion model, while sham group was only isolated carotid artery. Infarct size of brain tissue was measured by TTC staining. Focal deficit score, Morris water maze test, the rotating beam test and Rotarod test were resepectively made to evaluate the animal behavior after injury. Results Adequate amounts of BMSCs were harvested by adherence screening method and subculture. Ischemic area of BMSC group （ （34.98± 12.49）% ） was significantly smaller than that of MCAO group （ （ 42.36± 9.41 ） % ） at 2nd day after injury （P〈 0.05 ）. Compared with MCAO group,focal deficit score of BMSC group reduced significantly at 3rd day after injury,and got to the most significant differences at 5th day after injury （P〈0.01） ; escape latency of BMSC group was significantly shortened at 7th day and 14th day after injury in Morris water maze test （P〈 0.05） , meanwhile time percentage, distance percentage in the target quadrant and the times corssing the platform were increased gradually after injury, and reached significant differences at 14th day after injury （P〈0.05） ;exercise time in Rotarod runner increased at every time point after injury, and reached most significant differences at 14th day after injury（P〈0.01 ） ; walking speed in the rotating beam test increased most significantly at 14th day after injury,meanwhile walking distance at 5th and 10th day after injury（P〈0.01 ）. Conclusion BMSC transplantation via carotid artery can significantly improve neural function, learning, balance and motor function after brain injury, which will be a new way of TBI therapy.