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目的探讨脑缺血后,外源性植入的骨髓基质干细胞(bone marrow stromal cells,BMSCs)在脑内的迁移和分化,及脑缺血后自体神经干细胞(neural stromal cells,NSCs)的动员情况。方法体外培养、扩增BMSCs 后,用绿色荧光染料PKH67标记BMSCs,通过静脉和脑内局部移植等途径将BMSCs植入大脑中动脉闭塞的SD大鼠脑内,10d后取材,免疫荧光检测BMSCs的分化、迁移情况;并比较假手术和脑缺血时脑内自体NSCs的数量。结果 (1)静脉移植组两侧脑半球中PKH67+细胞数分别是46．4±2．92和21．8±1．13／100mm2(P<0．05);(2)移植的BMSCs中40．31％表达神经元特异性标志;(3)在第8天时,脑缺血半球和非脑缺血半球Nestin+细胞数分别为 19．5±10．08和7±1．41,差异显著(P<0．05)。结论脑缺血可激活脑内自身的NSCs,使它们迁移至脑缺血半球并促进其修复;外源性BMSCs通过局部或静脉移植后可迁移到缺血脑组织附近,并且部分分化为神经元样细胞,表达神经元的特异性标志。
Objective To investigate the migration and differentiation of exogenous bone marrow stromal cells (BMSCs) after cerebral ischemia and the mobilization of autologous neural stem cells (NSCs) after cerebral ischemia . Methods After BMSCs were expanded in vitro, BMSCs were labeled with green fluorescent dye PKH67. BMSCs were implanted into the middle cerebral artery occluded SD rats by intravenous and intracerebral transplantation. After 10 days, BMSCs were harvested and immunofluorescently detected Differentiation and migration. The numbers of autologous NSCs in the brain during sham operation and cerebral ischemia were also compared. Results (1) The numbers of PKH67 + cells in the bilateral hemispheres of vein graft group were 46.4 ± 2.92 and 21.8 ± 1.13 / 100mm2, respectively (P <0.05); (2) .31% expressed neuron-specific markers; (3) At day 8, the number of Nestin + cells in cerebral ischemia and non-ischemic hemispheres were 19.5 ± 10.08 and 7 ± 1.41 respectively P <0.05). Conclusion Cerebral ischemia can activate its own NSCs, make them migrate to the ischemic hemisphere and promote their repair. Exogenous BMSCs can migrate to the vicinity of ischemic brain tissue by local or intravenous transplantation and partially differentiate into neurons Like cells, specific markers of neurons.