Regeneration of neurons and vessels using bone marrow cells and gene delivery

我们初步将年轻小鼠的骨髓细胞静脉移植到老年加速衰老小鼠（SAM）体内。我们通过水迷宫评估记忆功能，但是没有观察到记忆功能的改善。因此，我们认为，静脉注射年轻骨髓对防止衰老和进一步加强治疗（如有用的基因诱导和向脑或骨髓靶向给药）的效果甚微。然后，我们通过一种新的治疗策略，用FGF-2基因转染的间充质干细胞（MSCs）检测了大鼠短暂性大脑中动脉闭塞（MCAO）后神经功能改善的效果。单纯疱疹病毒1型（HSV-1）载体。与其他组相比，接受FGF-2修饰的MSC的中风动物表现出显著的功能恢复。MCAO后14天，仅FGF-2修饰的MSC治疗组的梗死体积显著减少。FGF-2修饰的MSC-t中的FGF-2产生 ...更多信息 MCAO后第3、7天，再灌注组脑组织中MDA含量显著高于其他组。施用FGF-2修饰的MSC强烈表达FGF-2蛋白，这通过ELISA证实。结论：与单纯MSCs移植相比，FGF-2基因修饰的MSCs与HSV-1载体联合移植能促进脑卒中后功能的恢复，肝细胞生长因子（HGF）也是促进脑卒中后组织修复的有效基因之一。我们介绍了一种新的策略，在大鼠短暂MCAO模型中将MSC和离体HGF基因转移与多突变HSV-1载体相结合。超急性处理时，仅MSC-HGF组与PBS组在第14天梗死面积差异有统计学意义（P < 0. 05），而两组在第14天梗死面积差异均有统计学意义（P < 0. 05）。超急性期移植后，MSC-HGF组缺血脑组织中HGF蛋白表达较其他组明显增强，并持续2周以上。此外，我们确定HGF表达的增加来自于基因修饰的MSC中转移的HGF基因。与其他组相比，MSC-HGF组缺血边界区（IBZ）凋亡阳性细胞百分比明显减少，而IBZ皮质区残存神经元百分比明显增加。目前的研究表明，联合治疗比单独MSC细胞治疗更有效，并且可以将治疗时间窗从超急性期延长到急性期。这些实验表明，改善效果主要是由于抗凋亡和神经元保护作用，没有证据表明神经元生成。另一种有效的治疗方法如骨髓内注射骨髓细胞等值得进一步研究。少

We preliminarily intravenously implanted bone marrow cells from young mouse to old senescence accelerated mouse (SAM). We evaluate the memory function by water maze, however, no improvement in memory function was observed. Therefore, intravenous administration of young bone marrow has little effect on preventing aging and further enhanced treatment such as useful gene induction and targeting administration to the brain or bone marrow.Then we examined the effects of neurological improvement after transient middle cerebral artery occlusion (MCAO) in rats by a novel therapeutic strategy with FGF-2 gene-transferred mesenchymal stromal cells (MSCs) by the herpes simplex virus type 1 (HSV-1) vector. The stroke animals receiving FGF-2-modified MSCs demonstrated significant functional recovery compared with the other groups. Fourteen days after the MCAO, there was a significant reduction in infarction volume only in FGF-2-modified MSC-treated group. FGF-2 production in the FGF-2-modified MSC-t … More reated brain was significantly higher compared with the other groups at 3 and 7 days after MCAO. Administrated FGF-2-modified MSCs strongly expressed the FGF-2 protein, which was proven by ELISA. In conclusion these datum suggested that the FGF-2 gene-modified MSCs with the HSV-1 vector can contribute to remarkable functional recovery after stroke compared with MSCs transplantation alone.The hepatocyte growth factor (HGF) is also one of the useful gene for tissue restoration. we introduce a new strategy combining MSCs and ex vivo HGF gene transferring with a multimutated HSV-1 vector in a rat transient MCAO model. The significant difference of infarction areas on day was detected only between the MSC-HGF group and the PBS group with the superacute treatment, but was detected among each group on day 14 with both transplantations. After the superacute transplantation, we detected abundant expression of HGF protein in the ischemic brain of the MSC-HGF group compared with others on day 1 after treatment, and it was maintained for at least 2 weeks. Furthermore, we determined that the increased expression of HGF was derived from the transferred HGF gene in gene-modified MSCs. The percentage of apoptosis-positive cells in the ischemic boundary zone (IBZ) was significantly decreased, while that of remaining neurons in the cortex of the IBZ was significantly increased in the MSC-HGF group compared with others. The present study shows that combined therapy is more therapeutically efficient than MSC cell therapy alone, and it may extend the therapeutic time window from superacute to acute phase.Following these experiments, the improving effect is mainly due to anti-apoptotic and neuron-protective effects and there were no evidence for neuron-generation. Another effective methods such as intra-bone marrow injection of bone marrow cell should be examined in the future. Less