神经营养素3对大鼠急性脊髓损伤后Fas表达影响.doc

神经营养素3对大鼠急性脊髓损伤后Fas表达影响 　 作者：郭树章 任先军 蒋涛 欧阳忠 【关键词】 脊髓损伤 摘 要：［目的］探讨神经营养素3(NT-3)对脊髓损伤保护作用的分子机制。［方法］105只SD大鼠随机分成3组：对照组(生理盐水组)，实验组(NT3组)，假手术组。用改良Allen’s WD法以30gcm致伤SD大鼠制作大鼠全瘫模型，经蛛网膜下腔导管于术后即刻、4、8、12、24h、3、7d注入NT3 20μl(含NT-3 200ng)，对照组在相同时间点给予等容积生理盐水，假手术组只打开椎板后蛛网膜下腔置管，不致伤，不给药。采用免疫组织化学方法检测Fas蛋白在脊髓神经元的表达变化情况。［结果］假手术组中Fas蛋白弱阳性表达，对照组中Fas蛋白4h即出现强阳性表达，24h达高峰，实验组与对照组相比Fas蛋白表达明显减少(Plt;0．01)。［结论］NT-3能通过抑制Fas蛋白的表达抑制脊髓损伤后神经元凋亡，从而保护损伤的脊髓组织，这可能是NT-3对脊髓损伤具有保护作用的机制之一。 　　关键词：神经营养素3；脊髓损伤；Fas 　　Abstract：［Objective］To study the protective mechanisms of neurotrophin-3 (NT-3)on spinal cord injury.［Method］using the method of Allen’s (WD) to make the animal model of acute spinal cord injury of rats, 105 SD rats were randomly divided into three groups: control group, experimental group and sham group. A thin plastic tube was situated in subarachnoid space below the injury level for perfusion. Rats in experimental group received NT-3 20μl(including NT-3 200ng) from the tube at 0, 4, 8, 12, 24h and 3d, 7d after injury; the saline-treated animals received the equal volume of normal saline at the same time as control. The animals in sham group only received opening vertebral plate and putting tube in subarachnoid space. The rats were sacrificed at 4,8,12,24h and 3, 7, 14d postinjury (n=5). The expression levels of Fas protein in rats spinal cord were detected by immunohistochemistry assay. ［Result］The levels of Fas protein in control group were significantly increased as compared with those in sham group, and the level reached peak at 24h after spinal cord injury. The levels of Fas protein in NT-3 group were significantly decreased as compared with those in control group.［Conclusion］NT-3 can protect spinal cord from injury in vivo. One of mechanisms is that inhibits abnormal expression of Fas protein,then inhibits apoptosis after spinal cord injury. 　　Key words：Neurotrophin3;Spinal cord injury; Fas 　　程序性细胞死亡(programmed cell death，PCD)亦称凋亡(apoptosis)是一种自然的生理过程，在维持细胞和脊髓发育以及排除异常细胞过程中起关键作用。近年来凋亡在脊髓继发性损