【作者】 徐锦芳；

【导师】 沈宏； 刘伟国；

【作者基本信息】 浙江大学 ， 外科学， 2002， 硕士

【摘要】 周围神经是由离开脑和脊髓的神经纤维在由结缔组织连接、包裹组成粗细不等的神经束的基础上，再由结缔组织进一步包裹而形成的(肉眼可见的)神经束的整合体。解剖学上包括了除第一、二脑神经以外的所有脑神经、神经根、脊神经节、节段神经、神经丛和肢体的主要神经干。周围神经损伤(Periphefal nerve injury，PNI)是很常见的创伤性疾患，在人类的生活、劳动过程中均能发生，尤其是随着现代工业、交通运输业的迅猛发展，工伤和交通事故使周围神经损伤成为临床上的常见病之一。 到目前为止，周围神经损伤和再生仍然是创伤外科所未能完全解决的问题之一。在当前显微外科技术高度发展的情况下，仍然缺乏让其感觉能力完全恢复的方法，单纯依靠改进外科技术来促进神经损伤后功能恢复已经越来越困难，手术以外的领域受到了重视，近几十年来很多学者以较大的兴趣探讨影响受损神经形态和功能恢复的微环境问题，找到多种方法和药物以促进周围神经损伤修复。 溃江大学刃士研贝生学位论文 本课题选用健康雌性大鼠65 ．q，通过手术钳夹造成大鼠的坐骨神经钳夹 损伤模型，病理分类属Sunderland的Ill度损伤。在随后连续一周每天局部注 射天然细胞生长调控因子（NCGCF），间隔不同时间对坐骨神经进行电生理功 能指标观察，包括神经传导速度、潜伏期、动作电住峰值等。并取坐骨神经 0 制成石蜡标本切片，HE染色后光镜下观察神经病理生理形态，神经再生情况 等，做图象分析。通过与对照纽的比较，观察局部应用讹优F对周围神经再 生的影响，探讨其可能的作用机理。为临床上治疗周围神经损伤提供新方法 依据。 材斜与方法 1．动物分组：65只体重200－2309健康雌性SD大鼠随机分三组：＾组 （n－5）为假手术组，B组（n－30）为治疗组1，C组（n＝30）为治疗组2。 2．动物模型和手术：氯胺酮麻醉后，暴露好坐骨神经，无齿微血管钳钳 夹暴露好的神经段造成挤压伤模型。 3．给药方法和剂量：A组术前术后均不给任何药物。B组使用1号药物 皮下注射给药 0．Zml，术后每天皮T给药 0．Zml，连续 1周。C组使用 2号药 物，具体操作如B组。实验结束揭密：1号药为NCGCF，2号药为生理盐水。 4．实验结果的评价： （1）神经电生理检查：术后第 7、14、ZI、28、42天在 B组和C组的大 鼠中随机各取6 ．q，PCLAB多功能生物信号采集系统行电生理检查。i己录刺 激脉冲到肌肉动作电位产生时间（潜伏期）、肌肉动作电位峰值。求出神经千 传导速度（NCV）。 3 浙江大学项士研究生学位论文 （2）组织形态学观察：术后定期观察大鼠的一般情况，前述电生理检查 步骤完成后，取大鼠坐骨神经行神经组织学检查，HE染色，光镜下观察切片 中神经轴突和髓鞘的组织学变化。 5．数据统计和处理： 实验数据用均数土标准差（x i s）表示，以 SPSS10．0 for Windows进行 t检验。 ＃＄ 一、组织形态学观察 两组即时损伤坐骨神经侧的下肢功能均受限，见伤侧下肢足趾缩拢、垂 落，拖曳步态爬行。术后1周受损侧下肢足根部的毛部分脱落，发生失神经 营养改变，电生理检查时肉眼可见两组大鼠神经损伤侧腿部肌肉俱呈暗红色， 而正常侧肌肉呈红色。两组实验动物损伤的坐骨神经在光镜下观察可见雪旺 细胞肿胀，胞质内空泡，蜂窝样管泡状结构，轴浆中空泡，细胞器结构模糊。 术后2周开始，两组大鼠的运动功能有所恢复，后跟毛缺失区已缩。J、， 肉眼观察无显著差别。光镜下逐渐可见到形态正常的雪旺细胞和神经纤维； 可见到细胞间质中的一些固有成分、纤维、毛细血管等，形态正常的神经结 构占多数。从整体来看，神经纤维数目实验组稍多于对照组。 术后一周以后两组大鼠的下肢功能明显改善，后跟毛缺失区已缩小、消 失，肉got察无显著差别。光镜下观察结构已基本接近于正常神经纤维组织， 整体上实验组与对照组无显著的差别。 4 浙江大学刃士研纪生学位论文 二、神经电生理检查 表1 两组间不同时问NCV比较（单位：m／s） NCGCF治疗组更多还原

【Abstract】 The peripheral nervous system (PNS) comprises all nervous tissue outside the brain and spinal cord. It consists of groups of neurones (ganglion cells), called ganglia, feltworks of nerve fibres, called plexuses, and bundles of parallel nerve fibres that form the nerves and nerve roots. One nerve fibre consists of an axon and its nerve sheath. Each axon in the peripheral nervous system is surrounded by a sheath of Schwann cells. An individual Schwann cell may surround the axon for several hundred micrometers, and it may, in the case of unmyelinated nerve fibers, surround up to 30 separate axons. The axons are housed within infoldings of the Schwann cell cytoplasm and cell membrane.With the development of the modern industry and transportation, the incidence of peripheral nerve injury (PNI) increased recently. At the same time, the application of microsurgery technology increased the effects in the repair ofperipheral nerve injury. However, functional outcomes and peripheral nerve repair are usually unsatisfactory. It seems difficult to make a completely recovery in PNI patients with surgical treatment only. Other treatments besides microsurgical techniques such as neurotrophic factors and growth factors were thought highly Nowadays.In these studies, we used natural cell growth control factor (NCGCF) as a new rapid growth factor which is the very first time applied on peripheral nerve injury repair. Our research evaluated the histo-pathology and neural electric physiology of crushed sciatic nerve in rats. Compared the differences between the two groups, we studied the therapeutic effects of NCGCF on the crushed peripheral nerve and discuss its underlying mechanism.Materials and methods1. Group: 65 adult female SD rats weighted 200~-230g were randomly divided into 3 groups: Group A(n=5) as sham operation group. Group B(n=30) as treatment group 1. Group C(n=30) as treatment group 2.2. Surgical process and animal model: All rats were anesthetized with Ketamine Hydrocholoride. The sciatic nerve was exposed and crushed by a special flat-jawed microvascular clap.3. administration method: Group A underwent no crush or agent administration. Group B received injections No.l 0.2ml in volume by subcutaneous injection 24hand Ih prior to nerve crush and then daily for the following 7 days. Group C received injections injections No.2. After the end of the experiment, the injections were disclosured that No. 1 is NCGCF and No.2 is normal saline. 4.Evaluation of results:(1) Neural Electric physiology was examined on days 7,14,21,28, and 42 after nerve crush in all rats. The nerve conduct velocity (NCV) and the peak value of action potenial (AP) were evaluated by PCLAB system.(2) Histo-pathology changes were observed everyweek. After elecric procession, the sciatic nerve were cut off and underwent HE stain ,then observed under light scope.5.Data and statistics:All data in this article record as x眘 and processed by SPSS 10.0 for Windows, using independent T test.Result1. Histo-pathology:At the same time of crushed, the injuried leg was powerless. About 1 week later, the injuried leg had some hair lost on its knee and ankle. Under the light scope, the Schwann cells were swelling. Spongy structure and vague cell apparatus can be found. 2 weeks later, the hair lost region decreased. There neural fibers and Schwann cells were clear. Interstitials such as capillaries were foundunder light scope. 4 weeks later, the gross and histological appearances were all near normal structure. There were no signifkant differences between two groups.2. Neural Electric physiology:Table 1 NCV between two groups in different time (m/s)NCGCFNormal SalineTime ?PNCVnNCVn14d14.1 7 ?.99612.03 ?.0960.00521d22.60 ?.03619.18+1.6160.03528d37.55?.16635.27 + 1.0860.00642d43.75?.39642.13 + 2.3960.269Table1 AP peak betweentwogroups in different time (mv)NCGCFNormal SalineTime更多还原