【作者】 陈威；

【导师】 王冰；

【作者基本信息】 中南大学 ， 外科学， 2011， 硕士

【摘要】 目的探讨骨髓间充质干细胞移植治疗大鼠急性脊髓损伤的作用机制。方法1.运用改良Allen氏打击器制备大鼠T10脊髓损伤模型,假手术组48只,损伤组96只随机分为对照组和移植组,各组再分为移植术后1天、3天、7天、14天4个亚组,每亚组12只大鼠。对照组和移植组于损伤后7天分别经尾静脉分别注射磷酸盐缓冲液(PBS)1ml和含转绿色荧光蛋白的大鼠骨髓基质干细胞(GFP-BMSCs)单细胞PBS悬液1ml(1x106个BMSCs)。2.移植术后1天、3天、7天、14天,应用BBB评分法评价治疗前后损伤大鼠运动功能改善状况。3.移植术后14天,应用光镜及电子显微镜观察损伤脊髓组织形态学变化；移植术后3天、7天,应用荧光激发光技术观察BMSCs在体内迁移、存活情况。4.移植术后1天、3天、7天、14天,应用免疫组织化学技术检测损伤大鼠Nogo-A及SYN的表达。结果1.移植组与对照组损伤大鼠的运动功能均有不同程度的恢复。移植组损伤区脊髓结构与对照组相比,相对较完整,表现出较明显的运动功能改善(P<0.05)。2.移植组可见GFP标记的细胞在损伤区及其周边区明显聚集并存活。3.移植术后第3天、第7天和第14大,移植组脊髓损伤区周边组织中Nogo-A表达较对照组明显降低(P<0.05)。4.移植术后第3天、第7天和第14天,移植组脊髓损伤区周边组织中SYN表达较对照组明显升高(P<0.05)。结论1.BMSCs通过外周血管移植能够穿透血脑屏障,向脊髓受损区域发生迁徙,并在其内存活、分化促进大鼠急性脊髓损伤的神经功能的恢复。2.BMSCs移植治疗急性脊髓损伤可抑制受损局部Nogo-A蛋白的表达,从而减少胶质瘢痕的形成,促进轴突的再生。3.BMSCs移植治疗急性脊髓损伤有利于受损局部突触素(SYN)表达增加,促进突触功能的恢复。更多还原

【Abstract】 Objectives To study therapeutic benefit and mechanism of intravenous administration of bone marrow mesenchymal stem cells after spinal cord injury in adult rats.Methods .Rat’s acute SCI model was established with modified Allen’s method。There are 48 in the sham-operated group and 96 rats in the injuried group. Then the injuried group was randomly divided into transplantation group and control group.There were 4 time points in each group,n=12 in each point. At 7days after SCI,5×l06 BMSCs (1ml) were injected into the transplantation group by tail vein and 1ml (PBS) were given to the control group by tail vein. BMSCs were labeled by GFP before transplantation。2.Functional out come meaurements using the Basso-Beatti-Bresneh-an Score were performed at the 1st d、3rd d,7th d and 14th d after transplantation.3.Light microscopy and electron microscopy were used to identify the morphology of the sections of injured spinal cord tissue at 14th d after transplantation. Laser scanning microscopy were used to identify migrati-on and survival BMSCs in vivo at the 3rd d and 7th d after transplantation.4.The expressions of Nogo-A and SYN were detected by immunohistoc-hemical method at the 1st d、3rd d,7th d and 14th d after transplantation.Results 1.neurological functions of transplantation group and control group all were recoveried in varying degrees,but neurological function in the transplantation group was higher than it in the control group (P＜0.05).2.GFP staining positive cells were obviously gathered and survived in the injury zone and its surrounding area at the transplantation group.3.the expressions of Nogo-A in the transplantation group was down-regulated more significantly than the control group at the 3rd d、7th d、14th d after transplantation.(P<0.05)4.the expressions of SYN in the transplantation group was higher significantly than the control group at the 3rd d、7th d、14th d after transplantation.(P<0.05)Conclusion1.BMSCs transplanted by vein can penetrate through the blood-brain barrier, to spinal cord injury, and regional occurred in vitro migrating live, differentiation promote rat acute spinal cord injury nerve function recovery o2.BMSCs transplantation in the treatment of acute spinal cord injury can inhibit the damaged local Nogo-A protein expression, thus reducing glial scar formation, promote the regeneration of axons.3.Transplantation in the treatment of acute spinal cord injury BMSCs to local SYN expressing increasing, promote synaptic function recovery.更多还原