摘要
[目的]在大鼠急性脊髓损伤后不同时间硬膜外腔注射促红细胞生成素,观察并探讨其对脊髓神经功能的保护作用以及对脊髓神经细胞凋亡的影响。[方法]将48只体重(270±10)g成年雄性Sprague-Dawley大鼠随机分为正常组(n=6),假手术组(仅切除椎板,n=6),对照组(1、6、24h组分别给与等量生理盐水硬膜外注射,n=18),实验组(1、6、24h组分别给与5000u/kg-bw的rhEPO硬膜外注射,n=18)。按改良Allen打击法建立大鼠脊髓不完全损伤模型,通过动物神经运动功能BBB评分及斜板试验评价神经损伤程度;分别采用TUNEL法及免疫组化法检测脊髓神经细胞凋亡指数和Caspase-3,fas-fasl死亡蛋白的表达。光镜下观察统计分析结果。[结果]与对照组相比,脊髓损伤后促红细胞生成素1、6、24h干预的实验组神经运动功能均有改善,脊髓神经细胞凋亡指数均明显下降(P<0.01);实验组中,fas-fasl死亡蛋白较对照组表达减少。2组中神经细胞凋亡数与Caspase-3、fas-fasl死亡蛋白表达细胞数呈线性正相关(P<0.01),相关系数分别为r=0.941(Caspase-3)和r=0.929(fas-fasl)。[结论]早期应用外源性EPO对脊髓不完全损伤具有保护作用,可明显改善神经运动功能恢复情况,此种保护作用可能与EPO能够阻断Caspase-3、fas-fasl死亡蛋白途径的神经细胞凋亡有关。
[ Objective] To investigate the protective effects of rhEPO on neuromotor function and apoptosis of neural cell, after it was peritoneally injected at different time in rats with traumatic spinal cord injury（SCI）. [ Method] A total of 48 adult male Sprague-Dawley rats weighting 270 ＋ 10 g were randomly divided into four groups. Rats in normal group（ n = 6 ）and sham group （n = 6） underwent laminectomy producure only. In control group, rats （n = 18 ） received normal saline epidurally at 1,6 and 24 hours after injury. Treatment group （ n = 18 ） received 5000 iu/kg body weight of recombinant humane erythropoietin administered epidurally at l ,6 and 24 hours after injury. SCI was induced with 70g/cm impact according to the improved Allen method . Behavioral evaluation of the rats was made 48 hours after trauma using the Basso, Beattie, and Bresnahan （BBB） scoring system and Rivlin＇ s tihboard experiment. Injured spinal cord tissue cell apoptosis was examined by the terminal deoxynucleotidal transferasemediated dUTP-biotin nick end abeling （TUNEL） reaction at 4g hour. Fasl and Caspase-3 expression was determined by immunohistochemical analysis at 48 hours after injury. The results were observed by light microscope and analyzed by SPSS statistics software. [ Result] Compared to the control group, neuromotor function was significantly improved at 1,6 ,and 24 hours after injury in the experiment group. The indexes of neural cell decreased significantly （ P 〈 0.01 ）. Fasl and Caspase-3 expressed less in the experiment group. The number of neural cells was posibively correlated with the number of Caspase-3 （ r = 0.941 ）and Fasl （ r =0. 929） expressed cells（ P 〈 0.01 ）. [ Conclusion] Analysis of the results indicated that apoptosis plays an important role in the secondary spinal cord injury. Activation of Fasl and Caspase-3 may be related to cells apoptosis in the injured spinal cord. Early application of rhEPO could improve the neuromotor function. The protection is partially due to the reduction of neural cell apoptosis.
出处
《中国矫形外科杂志》
CAS
CSCD
北大核心
2008年第18期1419-1423,共5页
Orthopedic Journal of China
基金
江苏省卫生厅医学医学科研项目基金(编号:H200718)
苏州市社会发展项目基金(编号ssy0625)
