嗅鞘细胞移植联合跑步训练对改善脊髓损伤大鼠后肢运动功能的研究  

Study of olfactory ensheathing cells transplantation and treadmill training on improving hindlimb motor function of spinal cord injury rats

作　　者：王放[1] 高中洋[1] 张廷[1] 徐思越[1] 王栋[1] 李浩鹏[1] 贺西京[1] 

机构地区：[1]西安交通大学第二附属医院骨科,陕西西安710004

出　　处：《中国骨伤》2016年第10期928-938,共11页China Journal of Orthopaedics and Traumatology

摘　　要：目的 :研究嗅鞘细胞移植联合跑步训练对改善脊髓损伤大鼠后肢运动功能的效果,同时探索嗅鞘细胞移植联合跑步训练改善大鼠运动功能的可能机制。方法:选用雄性SD大鼠80只,随机分为4组,每组20只,均进行脊髓损伤造模后进行如下分组处理:生理盐水注射组(A组)、嗅鞘细胞移植组(B组)、生理盐水注射联合跑步训练组(C组)和嗅鞘细胞移植联合跑步训练组(D组)。建模3 d后C、D两组进行跑步训练,1周后B、D两组进行嗅鞘细胞移植(每只大鼠注射总量为4μl,细胞浓度为1.0×106/μl),A、C两组给予相应剂量盐水。观察时间为4周,每组每周测量BBB评分,利用免疫组化染色观察Bax、Bcl-2及NF-200的表达,采用Mallory磷钨酸苏木素染色观察神经纤维数量,使用TUNEL染色观察神经元凋亡。结果:(1)BBB评分:嗅鞘细胞联合跑步训练组与其他3组在第4周差异有统计学意义(P<0.05)。(2)在Bcl-2蛋白表达上,嗅鞘细胞移植联合跑步训练有交互作用,两者相互促进(P<0.05);在Bax蛋白的表达上,嗅鞘细胞移植可以明显降低其表达,与跑步训练交互作用不明显(P>0.05);TUNEL染色显示,嗅鞘细胞移植、跑步训练与时间因素三者具有交互作用,显著抑制细胞凋亡(P<0.05)。(3)在Mallory磷钨酸苏木素染色及NF-200免疫组化染色上,嗅鞘细胞移植、跑步训练与时间因素三者具有交互作用(P<0.05),促进神经再生。结论 :嗅鞘细胞移植联合跑步训练能够显著改善大鼠后肢运动功能,这可能通过以下途径实现:嗅鞘细胞移植与跑步训练能够相互协同,明显上调Bcl-2基因的表达,从而显著抑制神经元凋亡;同时能明显促进神经元轴索再生,提高神经纤维数量,并且这种效果可以随时间的延长更加显著。Objective：To investigate the effects of olfactory ensheathing cells （OECs） transplantation and treadmill training in improving hindlimb motor function of spinal cord injury （SCI） rats and explore its possible mechanism.Methods：A total of 80 male SD rats were randomly divided into four groups：media-untrained （group A）,OECs-untrained （group B）,mediatrained （group C） and OECs-trained（group D）.Each group had 20 rats and all rats were made the model of spinal cord injury.The rats of group C and D underwent treadmill exercise in 3 days after operation,the rats of group B and D underwent OECs transplantation in 7 days after operation （each rat was injected a total of 4 μl,cell concentration was 1.0× 106/μl）,at the same time,the rats of group A received the corresponding dose of saline.Then observed was continued for 4 weeks totally.BBB scores in each group were measured weekly.The expression of Bax,Bcl-2,NF-200 were observed by immunohistochemisty staining.Mallory staining was used to the regeneration of nerve fibers and TUNEL staining was used to observe neuronal apoptosis.Results：（1）The BBB scores in group D in the 4th week was significantly higher than of other three groups （P＜0.05）.（2） On the Bcl-2 protein expression,there was interaction between OECs and treadmill training,the two mutually reinforcing;while OECs transplantation can significantly reduce the Bax protein expression without significant interaction with treadmill training （P＞0.05）.TUNEL staining showed that OECs transplantation,treadmill training,the time factor had an interaction and significantly inhibited the apoptosis （P＜0.05）.（3）It was showed in immunohistochemisty staining of NF-200 and Mallory staining that OECs transplantation,treadmill training with the time factor were an interaction among the three with significant regeneration of nerve fibers （P＜O.05）.Conclusion：OECs transplantation combined with treadmill training can significantly improve hindlimb motor function o

关 键 词：脊髓损伤 嗅鞘细胞移植 跑步训练 细胞凋亡 神经再生 

分 类 号：R651.2[医药卫生—外科学]