脊髓内移植神经干细胞对臂丛神经根性撕脱伤后前角运动神经元生存和修复的影响  

Effects of intraspinal transplantation of neural stem cells on survival and repair of anterior horn motoneurons after brachial plexus avulsion

作　　者：涂致远 张文明[2] 朱维钦[2] 郑志 胡建石[3] 

机构地区：[1]武装警察部队福建总队医院骨科,福建省福州市350005 [2]福建医科大学附属第一医院骨科,福建省福州市350005 [3]福建医科大学分子生物学中心,福建省福州市350004

出　　处：《中国临床康复》2005年第26期82-85,共4页Chinese Journal of Clinical Rehabilitation

基　　金：福建省自然科学基金资助项目(C0210010)~~

摘　　要：目的:观察臂丛根性撕脱伤后脊髓内移植神经干细胞的存活、分化、迁移情况以及对原有前角运动神经元的影响。方法:实验于2004-01/12在福建医科大学分子生物学中心以及福建省骨科研究所完成。将雄性SD大鼠60只随机分为3组,每组20只。①单纯组:将C5～7神经根经后路撕脱制作大鼠臂丛根性撕脱伤模型,不进行细胞移植。②对照组:模型制作后即刻移植灭活神经干细胞(5×108L-1)4μL于C6脊髓前角。③实验组:模型制作后即刻把体外培养的5-溴尿嘧啶标记的神经干细胞(5×108L-1)4μL移植于C6脊髓前角。各组于术后1,2,4,8,12周5个时间点取脊髓标本制备切片,每个时间点4只,分别计算损伤侧脊髓前角运动神经元的存活率。观察移植神经干细胞存活、迁移、分化情况采用免疫组织化学染色方法。结果:60只大鼠全部进入结果分析。①各组大鼠臂丛根性撕脱伤后不同时间前角运动神经元存活率:实验组2,4,8,12周时间点均高于对照组和单纯组犤实验组:(79.3±2.9)%,(66.2±3.8)%,(57.0±5.4)%,(47.6±4.8)%;对照组:(69.4±3.1)%,(45.4±3.7)%,(33.4±6.5)%,(29.2±2.3)%;单纯组:(71.0±3.0)%,(45.9±2.9)%,(35.5±5.4)%,(27.9±1.9)%,t=3.8730～8.5009P<0.01犦。②神经干细胞移植入脊髓后不仅能存活,迁移达一个脊髓节段,并可以进一步分化为神经元以及星型胶质细胞。结论:①臂丛神经根性撕脱伤后脊髓前角内可以提供神经干细胞分化成星型胶质细胞和神经元的适宜微环境,而随时间不同表现出不同的分化趋向。②移植神经干细胞大鼠前角运动神经元存活率增高,说明神经干细胞移植除能分化为神经元而发挥神经元替代作用外,对神经根撕脱引起脊髓运动神经元死亡也具有保护作用,能明显减少神经根撕脱后运动神经元的继发性变性死亡。AIM： To study the survival, differentiation and migration of the transplanted neural stem cells into spinal cord, and the protective effect for the stem cells on anterior horn motoneurons after brachial plexus avulsion. METHODS： This experiment was conducted in the Center for Molecular Biology, Fujian Medical University and the Orthopaedic Research Institute of Fujian Province from January to December 2004. Sixty adult male SD rats were randomly divided into three groups and 20 rats in each group. ① Simplex control group did not receive transplantation after posterior nerve roots（C5 to C7） were avulsed to make rat brachial plexus avulsion model; ② Control group was transplanted with dead stem cells（5×10^8 L^-1, 4 μL） into C6 spinal cord anterior horn immediately after modeling of brachial plexus avulsion; ③Experiment group was transplanted with live neural stem cells （5×10^8 L^-1, 4μL） which had been cultured in vitro and labeled by BrdU before transplantation into C6 spinal cord anterior horn immediately after modeling of braehial plexus avulsion. At 1, 2, 4, 8, 12 weeks after operation, 4 rats in each group were perfused and spinal cords were dissected out to slice. The survival rate of the motoneurons in injured spinal cord anterior horn in each group was calculated in the three groups. The survival, migration and differentiation of the transplanted neural stem cells were observed with immunohistochemical staining. RESULTS： All the 60 rats were involved in the result analysis. ①Tbe survival rate of anterior horn motoneurons after rat brachial plexus avulsion was higher in experiment group than in the simplex control group and control group at 2, 4, 8, 12 weeks[（79.3±2.9）%, （66.2±3.8）%, （57.0±5.4） %, （47.6±4.8）% in the experiment group vs （69.4±3.1）%, （45.4±3.7）%, （33.4±6.5）%, （29.2±2.3）% in the control group and （71.0±3.0）%, （45.9±2.9）%, （35.5±5.4）%, （27.9±1.9）% in the simplex control group, t=3.873 0 to 8.500 9, P 〈 0.

关 键 词：臂丛/损伤 运动神经元 细胞死亡 干细胞 脊髓前角运动神经元 臂丛神经根性撕脱伤 移植神经干细胞 运动神经元生存 脊髓内 神经干细胞分化 

分 类 号：R651.3[医药卫生—外科学] R651.2[医药卫生—临床医学]