促红细胞生成素在脑红蛋白基因修饰的骨髓间充质干细胞向大鼠脊髓损伤部位迁移中的作用  

Role of erythropoietin in the migration of Ngb gene - modified bone marrow mesenchymal stem cells to the sites of spinal cord injury

作　　者：余化龙[1] 李军[1] 何宁[1] 刘志刚[1] 王志勇[1] 熊敏[1] 

机构地区：[1]湖北医药学院附属东风总医院骨外科,十堰442008

出　　处：《中华实验外科杂志》2016年第1期185-187,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省教育厅指导项目（B2015470）

摘　　要：目的观察促红细胞生成素（EPO）在脑红蛋白（Ngb）基因修饰的骨髓间充质干细胞（BMSCs）向脊髓损伤部位迁移中的作用。方法将96只成年SD大鼠随机分为4组：A：培养基组；B：EPO组；C：Ngb基因修饰的BMSCs组；D：EPO＋Ngb基因修饰的BMSCs组。脊髓损伤后A组静脉内注射DMEM培养基1ml，B组肌肉注射rhEPO（5000U／kg），C组静脉注射Ng4）基因修饰的BMSCs悬液1ml，D组肌肉注射rhEPO（5000U／kg）及静脉注射Ngb基因修饰的BMSCs悬液1ml。分别于术后第1、7、14、21天行Basso．Beatti．Bresnahan（BBB）运动功能评估，利用实时定量聚合酶链反应（Real-time PCR）检测Nsb基因的表达，通过原位缺口末端标记法（TUNEL）试剂盒检测损伤骨髓中细胞凋亡。结果术后第7天各组BBB评分分别为：5．14±0．63、8．36±0．37、8．45±0．52、9．92±0．73；术后第14天各组BBB评分分别为：7．58±0．94、10．39±0．89、11．12±0．78、14．21±1．06；术后第21天各组BBB评分分别为：8．59±0．77、14．12±0．99、14．33±1．61、18．02±1．78，D组运动功能改善显著优于其他各组，差异有统计学意义（P〈0．05）。损伤后各个时间点D组与C组NgbmRNA表达水平高于A组和B组，差异有统计学意义（P〈0．05）；C组与D组NgbmRNA表达水平差异无统计学意义（P〉0．05）。D组各个时间点细胞凋亡率低于B组和C组，差异有统计学意义（P〈0．05）；D组细胞凋亡率明显低于DMEM组，差异有统计学意义（P〈0．01）。结论促红细胞生成素能够促进脑红蛋白基因修饰的骨髓问充质干细胞的向脊髓损伤部位迁移，抑制细胞凋亡并促进脊髓损伤后的神经功能恢复。Objective To investigate the role of erythropoietin （EPO） in the migration of Ngb gene -modified bone marrow mesenchymal stem cells （BMSCs） to the sites of spinal cord injury. Methods Ninety - six adult SD rats were randomly divided into 4 groups ： A ： DMEM group; B ： EPO group; C ： Ngb gene - modified BMSCs group ; D ： EPO ± Ngb gene - modified BMSCs group. After spinal cord injury, 1 ml DMEM medium was intravenously injected in group A, rhEPO （5 000 U/kg） was intramuscularly injected in group B, 1 ml Ngb gene -modified BMSCs suspension was intravenously injected in group C, and rhEPO and Ngb gene - modified BMSCs suspension was injected in group D. Movement function was e- valuated through Basso- Beatti- Bresnahan （BBB） score on the postoperative day 1,7, 14, 21. The expression of Ngb gene was detected using real - time quantitative polymerase chain reaction （ Real - time PCR）. Cell apoptosis in the bone marrow tissue was detected using TdT - mediated dUTP nick end labeling （TUNEL） kit. Results At 7th day after spinal cord injury, BBB scores in group D were higher than those in group B and group C at each time point with the difference being statistically significant （P 〈 0. 05 ）. BBB scores in group D were Significantly higher than those in group A with significant difference （P 〈 0. 01 ）. BBB scores in groups A, B, C and D were 5.14 ±0. 63, 8. 36 ±0. 37, 8.45 ±0. 52, 9. 92 ±0. 73 seperately on the 7th day post - injury, 7.58 ±0. 94, 10. 39 ± 0. 89, 11.12 ±0. 78, 14. 21 ± 1.06 seperately on the 14th day and 8. 59 ±0. 77, 14. 12 ±0. 99, 14. 33 ± 1.61, 18.02 ± 1.78 seperately on the 28th day. Significant improvement was detected in group D as compared with other groups （P 〈 0. 05 ）. Ngh mRNA expression levels were higher in group D and group C than that in group A and group B at each time point with the difference being statistically significant （ P 〈 0.05 ） , and no significant difference was detected between group C and group D （ P 〉 0

关 键 词：促红细胞生成素 脑红蛋白 骨髓间充质干细胞 脊髓损伤 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]