人脐带间充质干细胞移植对急性脊髓损伤修复的实验效果观察

The transplantation of human umbilical cord mesenchymal stem cells has neuroreparable effect to injured spinal cord

目的通过GFAP的表达证明人脐带间充质干细胞在损伤脊髓内具有神经修复作用。方法制作36只wistar雌性大鼠脊髓损伤模型,随机为2组,每组18只:A组为对照组,行脊髓暴露手术,进行打击损伤,不做任何的移植; B组为干细胞移植组,脊髓损伤后经蛛网膜下腔进行人脐带干细胞移植。术后2M(月)行免疫荧光染色和免疫组织化学染色,每只老鼠切取损伤脊髓中心冰冻切片6片,荧光染色和组织化学染色间隔选片各3片,观察胶质纤维酸性蛋白(glial fibrillary acidic protein:GFAP)在大鼠损伤脊髓内表达以及迁移和分化等,两组分别在术后第1d、1周、3周、5周、7周、9周进行BBB行为学运动功能评分及GFAP在损伤脊髓内的表达,并利用统计学进行分析,了解损伤后的脊髓神经修复情况。结果 (1)术后各时段BBB评分,A组为(0. 00±0. 00); B组为(18. 33±0. 79)。(2)免疫组化显示:A组见GFAP少量阳性表达; B组见GFAP明显阳性表达。荧光染色显示:A组见GFAP少量阳性表达,B组见GFAP明显阳性表达。(3)统计学分析,A、B两组有显著差异(P <0. 05)。结论通过GFAP在损伤脊髓内的明显表达,表明人脐带间充质干细胞对急性脊髓损伤有较好的神经修复作用。

Objective：To prove that Human umbilical cord Mesenchymal Stem Cells has the faction of neurorepair through the expression of the GFAP in the injured spinal cord. Methods ： Making 36 models of wistar rat with injured spinal cord, then divide it into 2 groups at random, with every one 18 wistar rat. Group A is the control group with spinal cord undergoing exposure surgery, doing strike damage without any transplantation . Group B is the experimental group given the Human umbilical Stem Cells by Subarachnoid space transplantation after strike. All models were carried out Immunofluorescent staining and hnmunohistochemical staining two months later. Every rat was cut 6 chops of the frozen section out injured spinal cord at random with 3 pieces in hnmunofluorescent staining and the others in hnmunohistochemical staining. We judge the repair function of the injured spinal through the expression, transference and differentiation of the glial fibrillary acidic protein（GFAP）. Every group was Carried out BBB behavioral science movement function score and observe the expression in injured spinal cord after postoperative 1 d, 1 w, 3 w, 5 w, 7 w, 9 w to estimate the neural function recovery and analyse the data by the statistics. Results：（1）BBB score in each period , group A （00. 00 ±0. 00） ,there is no change. Group B （ 18.33 ± 0. 79）. （2）Immunohistochemical display： we find a small number of positive expression in group A while GFAP express clearly positively in group B. Fluorescent staining showed： group A express a small number of positive, with group B founding a clearly positive expression. （3）Statistical analysis demonstrate there is a significant difference （P 〈 0. 05 ） between Group A and B. Conclusion ： GFAP express positively in the injured spinal cord, it proves that Human um bilical cord mesenchymal stem cells has better effect of neurorepair for acute injured spinal cord.