血管内皮祖细胞移植治疗脊髓损伤的实验研究

目的 观察人的血管内皮祖细胞移植,对大鼠脊髓损伤的神经功能恢复的影响及移植细胞的存活和分化.方法 取由上海市发育生物学重点实验室提供的人血管内皮祖细胞,收集细胞悬液.将40只SD大鼠制备脊髓全横断模型,将存活的30只大鼠随机分为三组:假手术组(A组),10只,未做任何处理;手术/细胞组(B组),lO只,于距损伤区域1 cm椎管内注射7.5 μl,细胞总数为6×106个;手术/DMEM组(C组),10只,按B组方法注射等量DMEN.术后1、2,4、6和8周采用BBB后肢功能评分观察大鼠脊髓神经功能恢复情况,取出损伤的脊髓行免疫组织化学和原位杂交检测移植细胞在宿主脊髓内的存活和分化情况,再用SAS.1.3统计软件,分析行为学的变化,用秩和的Kruskal-Wallis Test和Nemenyi法检验,P<0.05为差异具有统计学意义.结果 BBB后肢功能评分显示术后各时间点A组与B、C组比较,差异均有统计学意义(P<0.05),B组与C组比较,在2、4、6和8周显示B组明显优于C组,但明显低于A组,上述差异均有统计学意义(P<0.05).原位杂交和免疫组化检测显示移植的内皮祖细胞不仅能在宿主体内存活而且一些移植的细胞嵌合到宿主脊髓并分化为血管.结论 人内皮祖细胞移植后,能够在脊髓内存活、并分化为血管内皮细胞并能促进损伤脊髓功能的恢复.

Experimental study of transplantation of endothelial progenitor cells for treating spinal cord injury

Objective To investigate the effects of transplantatation human endothelial progenitor cells onneurological functional recovery from spinal cord injury model in rats,survival of transplanting cells and differentia-tion. Method The human endothelial progenitor cells were provided by Shanghai Developmental Biology Labora-tory.Forty SD rats were made for the animal model of spinal cord complete transection.Thirty survival SD rats wererandomly divided into three groups:sham operation gronp(group A, n = 10),operation/cell group(greup B, n =10) and operation/DMEM group (group C, n = 10).Suspension containing (hEPCs 6×106) was transplanted in-to the vertebral canal around injured spinal cord. In group C, equal volume of DMEM was injected insbead in the same way as in the group B. The BBB score was obtained 2,4,6,8 weeks after injection, Immunohistochemistry andin-situ hybridization were used to observe the cells survival and differentiation in the spinal cord. The BBB test wasperformed to study the functional improvement of cells. The SAS version. 1.3 software for statistics was to studyethology and functional improvement. The sum of ranks was checlced with Kruskal-Wallis Test and Nemenyi test.Results There were statistically significant differences in BBB scoring between group A and group B as well asgroup C after operation (P<0.05). The BBB score in group B was higher than that in group C after 2,4,6 and8 weeks,but lower than in group A. The hybridization in-situ and immunohistochemistry showed that transplantedcells survived for 8 weeks after transplantation and expressed specific characteristics for ancestral cell and differentiated into vascular endothelial cell (VEC). Conclusions After transplantation, hEPCs can survive, differenti-ate into vascular endothelial cell,and improve spinal cord function as compared with control group.