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目的探讨脐血间充质干细胞(MSCs)静脉移植治疗新生鼠缺氧缺血性脑损伤(HIBD)的可行性及其时效性。方法脐血MSCs移植使用前以4′,6-二脒基-2-苯吲哚盐酸(DAPI)体外标记。实验选用7日龄SD大鼠38只制备HIBD模型,死亡3只,余35只共分3组:空白对照组(n=11);移植1组(n=12),在HIBD后第2天经鼠尾静脉注入脐血MSCs;移植2组(n=12),在HIBD后1周开始移植。两组均于移植后第2天以及HIBD后2周分别随机将鼠处死、取脑,用于脑组织病理形态学观察,并取海马回区相同部位的缺血脑组织切片,荧光显微镜下观察DAPI阳性细胞数。结果移植2组,1周后缺血脑组织细胞外间隙缩小,细胞数明显增加,脑组织水肿已明显减轻,在大鼠病灶侧脑内,可见大量的DAPI阳性细胞向病灶区及周围迁移和扩散,没有明显的界限。而移植1组于移植后病灶侧脑内很少见到DAPI阳性脐血MSCs分布,其脑组织水肿程度及细胞外间隙的改善和细胞数目的增加也不明显。结论脐血MSCs移植治疗新生大鼠HIBD,能有效透过血脑屏障,在病灶脑组织周围迁移、扩散、整合;移植时间选择HIBD后1周时有良好疗效。移植治疗过程中未见植入反应和其他不良反应。
Objective To investigate the feasibility and timeliness of intravenous transplantation of cord blood mesenchymal stem cells (MSCs) for neonatal hypoxic-ischemic brain damage (HIBD). Methods Cord blood MSCs were labeled in vitro with 4 ’, 6-diamidino-2-phenylindole hydrochloride (DAPI) before use. Thirty-eight SD rats aged 7 days were used to prepare HIBD model. There were 3 deaths, of which 35 were divided into 3 groups: blank control group (n = 11), transplantation group 1 (n = 12) Umbilical cord blood MSCs were injected into the caudal vein of rats. Two groups were transplanted (n = 12) and transplanted one week after HIBD. The rats were sacrificed on the 2nd day after transplantation and 2 weeks after HIBD, respectively. The brains were removed for histopathological observation. The ischemic brain sections of the same site in the hippocampus were observed. The results were observed under a fluorescence microscope DAPI positive cells. Results After transplanted into the two groups, the extracellular space of the ischemic brain tissue was reduced and the number of cells was significantly increased after 1 week. Edema of the brain tissue was significantly reduced. In the side of the rat lesion, a large number of DAPI positive cells migrated to and around the lesion area Diffusion, no obvious boundaries. In transplantation group 1, the distribution of MSCs in DAPI-positive cord blood was seldom seen in the lesioned brain, and the degree of edema and the improvement of extracellular space and the increase of the number of cells were also not obvious. Conclusion The transplantation of cord blood MSCs for the treatment of HIBD in neonatal rats can effectively migrate through the blood-brain barrier and migrate, diffuse and integrate around the brain tissue of the lesion. The transplantation of HIBD has good curative effect after one week of HIBD. Implantation and other adverse reactions were not observed during the transplantation.