嗅鞘细胞和雪旺细胞混合移植对大鼠脊髓损伤修复的影响  

Effect of mixed transplantation of olfactory ensheathing cells and Schwanne cells on spinal cord repair after spinal cord injury in rats

作　　者：叶超群[1] 孙天胜[1] 蔡艳华[2] 岳峰[2] 刘彦[3] 

机构地区：[1]北京军区总医院骨科,北京市100700 [2]首都体育学院研究生部,北京市100088 [3]北京市虹天济神经科学研究院,100045

出　　处：《中国脊柱脊髓杂志》2009年第9期676-681,共6页Chinese Journal of Spine and Spinal Cord

基　　金：国家自然科学基金项目(批准号:30872604);北京市委组织部优秀人才培养计划项目(编号:20071D0502000266);北京市虹天济神经科学研究院科研基金资助项目

摘　　要：目的:探讨嗅鞘细胞和雪旺细胞混合移植对大鼠脊髓损伤修复的影响。方法:75±1d雌性SD大鼠35只,随机取30只分为实验组和对照组,每组15只,剩下5只作正常备用。实验组和对照组大鼠均制作成T10段脊髓损伤模型,实验组在造模后2周接受嗅鞘细胞和雪旺细胞混合移植,移植部位为距离损伤中心0.5mm处的头侧和尾侧的脊髓中线上,每个部位注射4个点,深度为1.75mm、1.25mm、1mm、0.5mm;每点移植0.5μl含嗅鞘细胞和雪旺细胞各0.5×105个的DMEM悬液;对照组在相同部位注射等量DMEM。造模后每周应用BBB评分观察大鼠后肢运动功能,移植后6周每组大鼠在大脑运动皮层后肢代表区注射10%生物素化葡聚糖胺(BDA),移植后8周取包含损伤部位(对照组)和移植部位(实验组)的脊髓进行HE染色、胶质纤维酸性蛋白(GFAP)/神经丝(NF)免疫荧光双染、生长相关蛋白-43(GAP-43)的检测及显微荧光照相。结果:造模后1～4周两组大鼠后肢BBB评分未见明显差异,造模后5～8周实验组大鼠后肢BBB评分较对照组高(P<0.05),9、10周时两组无明显差异。伤后10周(移植后8周)时,实验组大鼠脊髓损伤部位瘢痕和损伤范围较对照组小;实验组脊髓损伤部位NF阳性纤维计数为31±8.12根/切片,明显多于对照组的17.80±2.58根/切片(P<0.01);实验组脊髓损伤部位GAP-43的相对表达量为1.27±0.12,明显高于对照组的1.20±0.58(P<0.05);两组损伤部位尾侧脊髓均未见BDA标记的皮质脊髓束。结论:嗅鞘细胞和雪旺细胞混合移植具有一定程度的促进大鼠损伤脊髓结构修复的作用,但未能促进大鼠后肢运动功能改善。Objective：To investigate the effect of mixed transplantation of olfactory ensheathing cells（OECs） and Schwanne eells（SCs） on spinal cord repair after spinal cord injury（SCI） in rats.Method：Aduh female SD rats（n=35,75±1d） were assigned into two groups（test group and control） randomly with each group 15,T10 SCI model was introduced by NYU impaetor（H=25mm）,and animals in test group received mixed transplantation of OECs and SCs（ 1：1 ） at four points located at midline of spinal cord 0.5mm rostrally and caudally to injured site at 2th weeks after injury （cell density ： 1 × 10^5/μl,injection depth ： 1.75mm, 1.5mm, 1.0mm, 0.5mm,injection dosage ：0.5μl cell suspension/per depth）（n=15 ）,while animals in control group received same doses of DMEM injection in the same site（n=15）.The open-field loeomotors rating scale of Basso,Beattie and Bresna- han（BBB score） was performed weekly from 1 week after injury to determine the functional recovery of hindlimbs.10% BDA was stereotaxically injected（n=6 per group） into bilateral motor cortex which control hind limb.The rats were executed 8th weeks after transplantation and the T8-T11 spinal cords which contained the complete lesion site or transplantation site were sectioned and stained by HE and Immunochemistry for fluo- rescent microscopy.Result：Compared with control group,the BBB score of hindlimbs of rats in test group showed great improvement between 5th-Sth weeks after injury （P〈0.05）,however no difference was noted between two groups from lth to 4th week and from 9th to 10th weeks after injury.At lOth week（8th week after transplantation） ,the scar area and injury site decreased.The number of neurofilament positive （NF＋） axons were 31±8.12/per section and 17.80±2.58/per section for test group and control group respectively,the intensity of growth associated protein-43 positively staining tissue（GAP-43＋） were 1.27±0.12/per section VS 1.20± 0.58/per section for test group and control gro

关 键 词：脊髓损伤 嗅鞘细胞 雪旺细胞 混合移植 BBB评分 轴突再生 大鼠 

分 类 号：Q813.1[生物学—生物工程] R683.2[医药卫生—骨科学]