不同途径移植HUCB-MSCs治疗脑血管病大鼠microPET-CT评价及其治疗机制研究

Currently, cerebral vascular disease is one of the three major causes to death in human disease, disability rates as high as 50%, and complications, and serious threat to human health. The human umbilical cord blood mesenchymal stem cells were studied in this topic. In vitro, amplification, marked respectively by the artery, vein, three-dimensional positioning after local way transplanted into ventricles of brain vascular diseases in rats through PET-CT observation on UCB-MSCs before and after transplantation and transplantation at different time points in rats with cerebral vascular disease model rats with brain injury changes of parts of blood flow and metabolism and nerve function score change scientific evaluation of curative effect of UCB-MSCs transplantation for treatment of cerebral vascular disease in rats. The UCB-MSCs in brain injury settlement, survival, differentiation, and apoptosis induced by secretion were detected. Expected results: The best way to clear cord blood stem cell transplantation was cleared; allogeneic MSCs during the wound healing process show a total of sources; UCB-MSCs may secrete a variety of nerve nutrition, growth factor; UCB-MSCs can protect neurons and receptors in brain tissue induced by neural stem cells differentiation, repair itself.

脑血管病其致残率高达50%-80%。本课题拟从脑细胞再生角度入手，研究人脐血间充质干细胞(human cord blood mononuclear cells UCB-MSCs)体外培养、扩增、标记后，分别经动、静脉及立体定位脑局部途径移植入脑血管病大鼠体内，通过小动物正电子发射断层显像（animal Positron emission tomography animal PET）观察UCB-MSCs移植前后及不同时间点该模型脑损伤部位血流及代谢变化、神经功能评分变化，评价UCB-MSCs移植的疗效；经免疫荧光组化、酶联免疫实验、rt-PCR检测UCB-MSCs在脑损伤部位定居、存活、分化、分泌、诱导、凋亡等情况，为临床细胞治疗提供依据。预期结果：明确移植的最佳途径；异基因MSCs在损伤修复过程中具有共源性；UCB-MSCs可分泌多种神经营养、生长因子及诱导自身神经干细胞分化、修复作用。

本课题从脑细胞再生角度入手，研究人脐血间充质干细胞(HUCB-MSCs)体外提纯、标记后，分别经动、静脉及脑局部途径移植入脑出血大鼠体内，通过micro PET-CT观察HUCB-MSCs移植前后不同时间点脑损伤部位代谢变化、神经功能评分变化及细胞归巢情况评估移植疗效；经ELISA、rt-PCR探讨其作用机制。结果：经各移植途径的模型大鼠在治疗后第三天可见HUCB-MNCS归巢至脑出血病灶及周边，三种途径HUCB-MSCs移植均可提高脑损伤部位代谢情况、改善大鼠的神经功能缺损情况。通过局部移植后rt-PCR检测人源及鼠源性NSE、MAP2发现细胞移植后表达量增加，说明HUCB-MSCs移植后可以促进内源性神经干细胞的再生。通过动脉移植后ELISA检测发现HUCB-MSCs移植后可提高脑内Ang-1表达水平，表明HUCB-MSCs可通过调节血管生成过程促进脑出血损伤区的恢复；通过静脉移植HUCB-MSCs后ELISA法检测炎性因子TNF-a及IL-6发现可抑制其表达，表明HUCB-MSCs可能通过减少炎性因子表达、抑制炎性反应发挥治疗作用。结论：细胞移植的最佳途径是立体定位脑局部途径，作用机制可能与促进内源性神经再生、血管新生及具有抗炎作用有关。