单纯感觉／运动神经纤维动物模型的制作及鉴定  

Generation and characterization of peripheral nerve animal model of pure motor/sensory nerve fibers

作　　者：彭建平[1] 寇玉辉[2] 邓玖旭[2] 张培训[2] 殷晓锋[2] 姜保国[2] 

机构地区：[1]上海交通大学医学院附属新华医院骨科,上海200092 [2]北京大学人民医院创伤骨科、北京大学交通医学中心,北京100044

出　　处：《北京大学学报（医学版）》2013年第5期807-814,共8页Journal of Peking University:Health Sciences

基　　金：国家杰出青年科学基金(30625036);国家自然科学基金(30801169、30971526、30973066);北京市科技新星(A-2008-10)项目资助~~

摘　　要：目的：制作单纯运动/感觉神经纤维动物模型并鉴定.方法：以SPF级SD大鼠为实验动物,随机分为感觉神经组（A组）、运动神经组（B组）、正常对照组（C组）,A组选择性切除L2~L4腹根,B组选择性切除L2~L4背根神经节、C组不做手术处理.术后2周、4周,取股四头肌行HE染色,腰髓行神经元核抗原（neuronal nuclei,NeuN）免疫组织化学染色,计数A组腰髓后角感觉神经元、B组腰髓前角运动神经元,股神经干分叉部分行锇酸染色,观察有髓神经纤维在皮支和肌支中的数量变化.结果：A组中大鼠右后肢股四头肌萎缩并逐渐加重,无自噬;B组中大鼠右后肢轻度肿胀,不同程度自噬,无肌肉萎缩.神经元计数：A组中,L2~L4段腰髓中平均每个10×20倍显微镜视野下感觉神经元计数分别为62.12±1.77（2周）、62.15±1.32（4周）;B组中,L2~L4段腰髓中平均每个10×20倍视野下前角运动神经元计数分别为30.12±0.44（2周）、30.00±1.87（4周）;C组中,平均每个10×20倍视野下后角感觉神经元计数为62.57±1.02,前角运动神经元计数为29.73±3.03;两个时间点与正常L2~L4段脊髓神经元计数差异无统计学意义.A组中,股神经肌支中部分神经纤维退变,皮支中无退变,有髓神经纤维计数为：1 558.17±50.14（2周）、1 544.00±47.42（4周）;B组中,股神经皮支神经纤维均退变、肌支中部分神经纤维退变,有髓神经纤维计数分别为387.67±48.50（2周）、393.50±27.86（4周）,2周和4周的计数结果比较差异无统计学意义.2周时退变的轴突及髓鞘尚未完全清除,4周时退变的轴突及髓鞘已完全清除.结论：通过选择性切除腹根/背根神经节可以获得单纯感觉神经/运动纤维动物模型.术后4周时,退变的轴突及髓鞘已完全清除,此时可获得良好的单纯性质周围神经,以此方法制作单纯感觉/运动神经纤维动物模型稳定可靠,可用于以后的实验研究中.Objective ：To generate peripheral sory nerve fibers, and identify them. Methods： into 3 groups. In group A, we ablated L2 -IA nerve animal model of pure motor nerve fibers/pure sen- The SPF SD rats were adopted in this study, and divided ventral roots （VRs） to generate peripheral nerve animal model of pure sensory fibers. In group B, we ablated L2 - L4 dorsal root ganglions （DRGs） to generate peripheral nerve animal model of pure motor fibers. Two time end-points were set as 2 weeks and 4 weeks. Neuron ceils in lumbar spinal cords were detected by immunohistochemical staining with antibody of neuronal nuclei （NeuN）. Motor neuron cells in lumbar spinal cords of pure motor fiber animal models and sensory neuron cells in lumbar spinal cords of pure sensory fiber animal models were counted respec- tively, and then compared to that of normal animals. Femoral nerves distal to the furcation were stained in osmium tetroxide, and then myelinated nerve fibers in the muscle branch and cutaneous branch of femoral nerve were counted respectively. Results： The mean numbers of sensory neuron cells and motor neuron cells in normal lumbar spinal cords were 62.57 ±1.02 and 29.73 ± 3.03 per 10 × 20 visual field respectively. For different end-points, the mean numbers of sensory neuron cells after ablating ventalfoots were 62.12 ± 1.77 （2 weeks）, 62.15± 1.32 （4 weeks） per 10 ×20 visual field respectively; the mean numbers of motor neuron ceils after ablating DRGs were 30.12 ±0.44 （2 weeks）, 30.00 ± 1.87 （4 weeks） per 10 × 20 visual field respectively. In group A, motor axons in muscle branch were degene- rated as the sensory axons in muscle branch and cutaneous branch were not changed. The senory axons in femoral nerve for the two end-points were 1 558. 17 ＋- 50. 14 （ 2 weeks） and 1 544.00 ± 47.42 （4 weeks）. In group B, sensory axons in muscle branch were degenerated as the motor axons were reserved. The motor axons in muscle branch for the two end-points were 387.67 ±48.50 （2

关 键 词：周围神经 神经纤维 神经再生 模型 动物 

分 类 号：R338[医药卫生—人体生理学]