利用腹壁反射建立人工膀胱反射弧的辣根过氧化物酶逆行示踪研究  

Experimental study on the artificial bladder reflex arc established by abdominal reflex after spinal cord injury with the HRP retrograde tracing method

作　　者：王金武[1] 侯春林[2] 李继峰[3] 陈爱民[2] 张伟[2] 王诗波[2] 

机构地区：[1]上海交通大学附属上海市第六人民医院骨科,上海市200233 [2]第二军医大学附属长征医院骨科,上海市200003 [3]复旦大学医学院手功能实验室,上海市200032

出　　处：《中华创伤骨科杂志》2004年第4期414-417,共4页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金(30170956);上海市博士后科研基金资助项目(01-1)

摘　　要：目的研究利用腹壁反射可建立“腹壁反射-脊髓中枢-膀胱”这一新的神经传导通路,通过再生到盆神经的躯体运动神经纤维可实现膀胱的神经再支配,探讨通过刺激下腹壁,该人工膀胱反射弧产生控制性排尿的神经形态学基础。方法选用6只SD大鼠,右侧为实验侧,左侧为对照侧,右侧T13前根近端与右侧S2前根远端通过一段自体移植神经在硬膜囊内行显微缝合,保持T13后根完整,经一段时间轴突再生后,建立“腹壁反射-脊髓中枢-膀胱”这一新的人工膀胱反射弧。神经缝合术后8个月,应用辣根过氧化物酶(peroxidasehorseradish,HRP)逆行追踪标记技术,用微量注射器在双侧膀胱壁上各注射30%HRP5μl,动物存活60h后经升主动脉灌注杀死。切取T11～L1及L4～S4节段脊髓,冰冻切片后利用TMB法进行呈色反应,观察切片内HRP标记细胞形态及分布情况。结果大鼠膀胱平滑肌注射HRP后,实验侧T13脊髓前角及双侧L6～S4节段脊髓中间带外侧核区有HRP标记阳性细胞,而对照侧T13脊髓前角未发现HRP标记细胞。结论腰骶髓截瘫平面以上的躯体运动神经轴突可长入膀胱平滑肌内副交感神经节细胞,并由此传递躯体反射冲动到膀胱平滑肌。利用“腹壁反射-脊髓中枢-膀胱”这一人工反射弧,对实现SCI患者膀胱控制性排尿功能,有一定临床意义。Objective To study the establishment of a new artificial reflex pa thway, “abdominal reflex-spinal cord center-bladder”and the reinnervation of bladder by the somatic motor nerve regeneration through the pelvic nerve, and t o explore the neural morphological basis of the artificial bladder reflex arc co ntrolling micturition induced by scratching the lower abdominal wall. Methods 6 SD rats were used, with the right side as the experimental side and the left sid e as the control side. The intradural microanastomosis of the right T13 ventral root to S2 ventral root was done with autogenous nerve grafting, leaving the rig ht T13 dorsal root intact. After axonal regeneration, the new artificial bladder reflex pathway, “abdominal reflex-spinal cord center-bladder”, was establis hed. 5μl of 30%horseradish peroxidase (HRP) solution was injected into either side of the corpus vesicae at 8 months postoperatively. The animals survived for 60 hours and were killed by perfusion through ascending aorta. The T11～L1 spin al segments and the L4～S4 spinal segments were sectioned with frozen section me thod, and treated with TMB method. The figure and distribution of HRP labeled ce lls in sections were observed under optical microscope. Results There were HRP l abeled cells in T13 ventral horn on the experimental sides and in the intermedio lateral nucleus on either side of the L6～S4 segments after HRP injection. There was no HRP labeled cell in T13 ventral horn on the control sides. Conclusions T he somatic motor axons above the lumbosacral paraplegia level can innervate blad der parasympathetic ganglion cells and thereby transfer somatic reflex activity to the bladder smooth muscle. The “abdominal reflex-spinal cord center-bladde r”artificial reflex pathway is practical in the SD rat model and may have a pot ential of clinical application with the function of controlling micturition for patients with spinal cord injury.

关 键 词：神经形态学 人工反射弧 脊髓损伤 人工膀胱 副交感神经 辣根过氧化物酶 腰骶髓损伤 

分 类 号：R651.2[医药卫生—外科学]