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1、脊髓损伤大鼠运动及神经功能自然恢复规律的探讨脊髓损伤大鼠运动及神经功能自然恢复规律的探讨王红星 徐冬晨 姚莉 雷晓婷 刘兴波 丁新生 王彤【摘要】目的观察不完全脊髓损伤（SCI）大鼠运动及神经功能自然恢复情况，为SCI后运动训练时机选择提供依据。 方法共选取45只成年SD大鼠，分为实验组（40只）和假手术组（5只）。实验组手术切除T10椎板暴露脊髓，采用改良Allens撞击法致SCI；假手术组仅手术切除T10椎板暴露脊髓。实验组分别于损伤前及损伤后1,3,5,7,14 d,21 d和28 d，假手术组分别于术前及术后1,3,5,7 d时采用斜板试验、改良Tarlov评分、BassoBeatti
2、eBresnahan（BBB）评分进行运动功能评定，采用脊髓诱发电位评定神经功能。实验组于上述各时间点分别取5只大鼠处死，假手术组于术后7 d时处死，取2组大鼠T10节段脊髓进行形态学检测。 结果实验组大鼠在损伤后13 d斜板角度、改良Tarlov评分和BBB评分均较损伤前显著降低，自损伤后5 d时开始增加，至14 d时达到平台期，显著高于术后1，3，5 d及7 d时水平（P0.05），但仍低于损伤前水平（P0.05）。实验组大鼠在损伤后1 d时脊髓体感诱发电位（SCEP）潜伏期较损伤前明显延长(P0.05）；波幅在损伤后1 d时明显降低，随时间进展呈逐渐增加趋势；假手术组术后各时间点潜伏期和
3、波幅与术前比较，差异均无统计学意义（P0.05）。2组大鼠术前脊髓结构完整，实验组术后13 d脊髓灰白质可见片状出血、细胞肿胀及变性；术后57 d炎性细胞减少，可见细胞内嗜碱性颗粒沉积、胶质细胞及少量神经纤维增生等；术后1428 d可见胶质细胞、神经纤维增生明显，细胞内有空泡结构形成；假手术组大鼠脊髓形态学方面手术前后无明显改变。 结论SCI大鼠运动功能、神经功能及脊髓病理形态学变化均与损伤时程密切相关，其运动功能改善一般于损伤后14 d时达到平台期，而神经功能改善一般于损伤后21 d时达到平台期。【关键词】脊髓损伤；运动功能；脊髓诱发电位Natural recovery of locomot
4、or and neurological function in rats after spinal cord injuryWANG Hongxing*, XU Dongchen, YAO Li, LEI Xiaoting, LIU Xingbo, DING Xinsheng, WANG Tong. *Department of Rehabilitation Medicine, The 1st Affiliated Hospital of Nanjing Medical University, Nanjing 210029, ChinaCorresponding author: WANG Ton
5、g, Email: 【Abstract】Objective To observe the natural recovery of locomotor and neurological functioning in rats after incomplete spinal cord injury. MethodsFortyfive SpragueDawley rats were divided into two groups: an experimental group and a shamoperation group. In the experimental group, the spina
6、l cord was exposed at the T10 level by laminectomy, and an animal model of spinal cord injury (SCI) was created using a modification of Allens method. The shamoperation group received only laminectomy without spinal cord injury. Locomotor and neurological functioning were evaluated using inclined pl
7、ane tests, modified Tarlov scores, BassoBeattieBresnahan (BBB) scales and spinal cord evoked potential (SCEP) before the operation and on the 1st, 3rd, 5th, 7th, 14th, 21st and 28th day post surgery. Histomorphological changes were observed by light microscopy. Results(1) In the experimental group,
8、the angles of the inclined plane test and the Tarlov and BBB scores had decreased significantly 1 and 3 days postinjury compared with those before the injury, and then increased gradually from the 5th day onwards, and peaking at the 14th day after injury. In the shamoperation group no significant ch
9、anges in the inclined plane test results, modified Tarlov scores or BBB scores were observed 1, 3, 5 or 7 days post surgery. (2) In experimental group, SCEP latency had increased significantly on the 1st day after SCI, but it then decreased gradually until reaching a plateau at the 21st day after th
10、e operation. The SCEP amplitude also decreased significantly at the 1st day after SCI, then increased gradually from the 3rd to the 21st day. In the shamoperation group, no significant changes in SCEP latency or amplitude were observed. (3) One and 3 days after the operation, hemorrhage and edema in
11、 the spinal cord as well as nerve fiber degeneration was observed in the experimental group. From day 5 to day 7, empty cavities formed gradually and astrocytes increased, a few nerve fibers grew. By days 14 and 28, more nerve fibers and astrocytes were found. ConclusionThe recovery of locomotor and
12、 neurologic function correlated closely with the natural course of spinal cord recovery, and becomes stable 14 to 21 days after injury. 【Key words】Spinal cord injury;Locomotor function;Somatosensory evoked potential脊髓损伤（spinal cord injury, SCI）是重要的致残原因之一。相关实验及临床观察发现，运动训练能有效促进SCI后功能恢复，是促进脊髓神经再生及功能恢复的
13、有效措施之一13。由于SCI后机体神经结构及功能变化复杂，难以确定运动训练的最佳介入时机。本研究通过观察SCI大鼠模型运动与神经功能自然恢复的时间规律，为SCI患者康复介入时机选择提供实验依据。现报道如下。材料与方法一、实验动物及分组共选取成年健康雄性SD大鼠55只，体重（32030）g，由上海斯莱克实验动物有限责任公司提供许可证号SCXK（沪）20030003。在模型制作过程中及术后共死亡10只被剔除，余45只纳入实验分析，将其分为实验组（40只）和假手术组（5只）。实验组大鼠行手术切除T10椎板暴露脊髓并致SCI，假手术组大鼠则仅手术切除T10椎板暴露脊髓组织。二、SCI模型制作过程参照文
14、献4介绍的方法，采用1%戊巴比妥钠（0.5 ml/100 g体重）腹腔注射麻醉成功后，将大鼠俯卧位固定，暴露T10椎骨棘突，切除椎板，暴露硬脊膜，然后将大鼠固定于立体定位器上，使脊柱处于水平位。采用改良Allens撞击法，在脊髓背侧面放置一金属垫片作为打击板，将10 g的砝码自4 cm高度垂直落下撞击打击板致大鼠SCI（术后大鼠尾巴痉挛摆动后双下肢瘫痪表明制模成功）。预防性应用青霉素进行肌肉注射（20万U，1次/d，共3 d）。实验组大鼠制模后每日上、下午各进行1次腹部按摩和挤压操作以协助排尿。三、运动功能评定实验组大鼠于损伤前、损伤后1，3，5，7 d，14，21 d及28 d时进行运动及神
15、经功能评定，假手术组大鼠于术前及术后1，3，5 d和7 d时进行功能评定。具体评定内容包括以下方面。1.斜板试验：将实验大鼠置于光滑木板上，其身体轴线与平板纵轴平行，平板每次升高5，以大鼠能停留5 s（期间保持不下滑）的最大角度为其功能值5。2. Tarlov评分：0级后肢无活动，不能负重；1级后肢可见活动，但不能负重；2级后肢活动频繁或有力，不能负重；3级后肢可支持体重，能走12步；4级可行走，仅有轻度障碍；5级行走功能正常6。3. BassoBeattieBresnahan（BBB）评分7：该评分系统将大鼠后肢运动功能分为22个等级，其中后肢全瘫为0分，功能完全正常为21分，主要观察内容包括关节活动数目和范围、负重程度及前后肢协调性、前后爪和尾部活动情况等。上述运动功能评定时间均统一为上午800。四、脊髓诱发电位检查参照文献8,9介绍的方法，采用Medtronic Keypoint 4ch型诱发电位仪检测大鼠脊髓体感诱发电位（spinal cord evoked potential，SCEP），电刺激强度为10 mA，波宽1 ms，频率2 Hz，叠加200次，将刺激电极正极插入大鼠后肢刺激胫后神经，负极位于正极远端2 cm处，记录电极则插于T8T9棘间韧带处，参考电极位于同
