大鼠自体嗅黏膜移植修复胸髓损伤:形态学和行为学的验证  

Transplantation of autologous olfactory mucosa in the restoration of spinal cord injury in rats Verification in morphology and praxiology

作　　者：陈庆山[1] 陈欣[2] 杨磊[1] 王珂[1] 石献忠[1] 孙宇[2] 潘胜发[2] 于恩华[1] 周长满[1] 

机构地区：[1]北京大学医学部人体解剖学与组织胚胎学系,北京市100083 [2]北京大学第三医院骨科,北京市100083

出　　处：《中国临床康复》2006年第13期8-10,i0001,共4页Chinese Journal of Clinical Rehabilitation

基　　金：北京大学医学部"985"项目基金(89100-612204001);北京市自然科学基金(7022018)~~

摘　　要：目的:采用自体嗅黏膜作为组织供体修复脊髓损伤,从形态学和行为学角度验证嗅成鞘细胞移植治疗脊髓损伤的作用及效果,为应用自体嗅成鞘细胞移植治疗临床脊髓损伤提供实验依据。方法:实验于2002-06/2004-10在北京大学基础医学院解剖学与组织胚胎学系中枢神经发育与再生研究室完成。选取清洁级6周龄雄性SD大鼠18只,随机分为自体嗅黏膜移植组10只,冻融对照组8只。两组均建立脊髓损伤模型,自体嗅黏膜移植组取位于鼻中隔后上部的嗅黏膜移植于脊髓损伤处,冻融对照组将自体嗅黏膜冷冻于-20℃冰箱5min,取出后室温放置5min解冻,重复5次后将冻融自体嗅黏膜移植于脊髓损伤处。术后6周于脊髓损伤节段上下各5mm处取材,片厚20μm。然后两组分别以神经丝蛋白和神经生长因子受体蛋白免疫荧光双标染色、激光共聚焦显微镜检测移植效果。免疫组织化学染色观察移植嗅黏膜与周围组织的生长情况。通过BBB行为学评分对两组动物后肢恢复功能进行评估。结果:实验选取SD大鼠18只,全部进入结果分析。①免疫组织化学摄片观察移植细胞在脊髓组织中再生情况:自体嗅黏膜移植组有部分空洞形成,但有部分神经纤维穿过移植区,可见神经生长因子受体蛋白免疫反应阳性的嗅成鞘细胞呈束状排列。冻融对照组无再生纤维通过且形成较大空洞,神经生长因子受体蛋白p75免疫组化反应阴性。②两组大鼠的神经丝蛋白和神经生长因子受体蛋白免疫荧光双标染色结果:自体嗅黏膜移植组免疫荧光标记纤维中夹有神经生长因子受体蛋白p75标记的存活的嗅成鞘细胞,细胞随标记的神经丝蛋白纤维走行方向排列,与宿主组织愈合良好。冻融对照组未见神经生长因子受体蛋白p75标记的嗅成鞘细胞以及神经丝蛋白标记的再生纤维,且形成较大空洞。③两组大鼠手术前后行为学评分结�AIM： To verify the effect of transplantation of sheath cells in the treatment of spinal cord injury （SCI） from morphology and praxiology by taking the autologous olfactory mucosa as tissue for restoring SCI, so as to provide the experimental basis for applying the transplantation of autologous sheath cells in treating clinical spinal cord injury. METHODS： The experiment was conducted in the Laboratory of Central Neural Development and Regeneration of Department of Human Anatomy, Histology and Embryology of Medical Faulty, Basic Medical College of Peking University from June 2002 to October 2004. Eighteen male SD rats of clean grade, aged of 6 weeks, were selected and randomized into autologous olfactory mucosa transplantation group （transplantation group） （n=10） and frozen-defrosted control group （control group） （n=8）. Rats in both groups were made into spinal cord injured models, and olfactory mucosa, which was in the top part of the back of anasl septum, of rats in the transplantation group were transplanted to the spinal cord injury, while that in the control group were frozed in refrigerator at 20 ℃ for 5 minutes, then took out for 5-minute-thawing. After 5 times repetition, the frozendefrosted autologous olfactory mucosa were transplanted into the spinal cord injury. Six weeks after the operation, materials were drew from both above and below segments of spinal cord injury for 5 mm with the thickness of 20 μm. Effects of both groups were assayed with neurofilament （NF） protein and nerve growth factor receptor protein immunofluorescent double labelled staining, laser confocal microscopy. The growing status of transplanted olfactory mucosa as well as its peripheral tissues were observed with immunonistochemical staining method, and the functional recovery of hind limbs in rats of beth groups were evaluated with BBS praxiologiy scale. RESULTS： Eighteen included SD rats in the experiment were involved in the analysis of results. ①Observation of the regeneration of transplan

关 键 词：脊髓损伤 嗅球/细胞学 神经生长因子 神经组织蛋白质类 神经纤维 神经再生 移植 自体 免疫组织化学 染色法 

分 类 号：R651.2[医药卫生—外科学]