【作者】 王东；

【导师】 骆文龙；

【作者基本信息】 重庆医科大学 ， 耳鼻咽喉科学， 2009， 硕士

【摘要】 背景研究证实,面神经损伤后,面神经元自身将合成鞘脂激活蛋白原促进神经元修复。目的观察模拟人体面神经损伤SD大鼠的面神经创伤模型面神经核鞘脂激活蛋白原阳性产物的时程变化及再生条件下的合成规律。设计、时间及地点神经病理学的随机对照动物实验,于2007-03/2008-09在重庆医科大学完成。材料成年雄性SD大鼠48只,随机分为面神经单纯切断组和面神经切断后即刻端端吻合组,每组24只。兔抗鼠Prosaposin多克隆抗体、即用型SABC免疫组化试剂盒、兔抗鼠Prosaposin一抗工作液(1:500)由武汉中美科技技术有限公司提供。方法面神经单纯切断组结扎左侧面神经耳后支远端主干,结扎点远端切除长约5 mm的神经干。面神经切断后即刻端端吻合组切断左侧面神经耳后支远端主干后,立即行神经外膜吻合。以两组动物右侧面神经作为正常对照。主要观察指标损伤后1,3,7,14,21,35d,以免疫组织化学方法检测两组大鼠面神经核中鞘脂激活蛋白原阳性神经元数量和免疫反应强度的变化。结果面神经单纯切断组:损伤侧面神经核鞘脂激活蛋白原阳性神经元的数量和免疫反应强度于术后第l天即开始增多,3d时升高明显,表达水平均高于对照侧,7d下降至最低。面神经切断后即刻端端吻合组:术后7d内损伤侧面神经核内鞘脂激活蛋白原阳性神经元数量和免疫反应强度变化规律与面神经单纯切断组相同,损伤侧面神经核内鞘脂激活蛋白原阳性神经元数量和免疫反应强度的回升最早出现在吻合后第14天,术后35d恢复接近至正常水平。术后14,21,35d面神经核鞘脂激活蛋白原阳性神经元的数量和免疫反应强度高于面神经单纯切断组(P<0.05),大鼠触须节律性拂动恢复,且大鼠触须节律性拂动恢复的时间与鞘脂激活蛋白原阳性神经元数量回升的时间基本一致。结论面神经损伤后7d内,面神经核鞘脂激活蛋白原表达呈先升后降的趋势,实施面神经修复对其表达影响不大。面神经损伤14d后,实施神经吻合可的表达。面神经核鞘脂激活蛋白原的回升与神经再支配同步出现。更多还原

【Abstract】 BACKGROUND Studies have demonstrated that damaged facial nerves synthesize prosaposin to promote repair of facial neurons.OBJECTIVE To observe time-course changes of prosaposin expression in the facial nerve nucleus of Sprague Dawley rats following facial nerve transection and repair.DESIGN, TIME AND SETTING A randomized control neuropathological animal experiment was performed in Chongqing Medical University between March 2007 and September 2008.MATERIALS A total of 48 adult, male, Sprague Dawley rats were selected and randomly divided into transection and transection + end-to-end anastomosis groups (n=24). Rabbit anti-rat prosaposin antibody, instant SABC immunohistochemical kit, and antibody dilution solution were purchased from Wuhan Uscn Sciences Co.,Ltd,China.METHODS In the transection group, the nerve trunk of the distal retroauricular branch of the left facial nerves was ligated in Sprague Dawley rats, and a 5-mm nerve trunk at the distal end of the ligation site was removed. In the transection + end-to-end anastomosis group, epineurial anastomosis was performed immediately following transection of the left facial nerves. The right facial nerves in the two groups served as the normal control group.MAIN OUTCOME MEASURES The number of prosaposin-positive neurons, as well as intensity of immunostaining in facial nerve nucleus, following transection and end-to-end anastomosis were determined by immunohistochemistry at 1, 3, 7, 14, 21, and 35 days after injury.RESULTS Transection group: transection of facial nerves resulted in increased number of prosaposin-positive neurons and immunoreactivity intensity in the facial nucleus on day 1. These values significantly increased by day 3. Expression was greater than in the control side. The peak of the reduction was reached at 7 days post-surgery. Transection + end-to-end anastomosis group: the number of prosaposin-positive neurons and immunoreactivity intensity was reduced in the facial nerve nucleus following immediate end-to-end anastomosis on day 7 post-surgery. These values began to gradually increase by day 14 post-anastomosis. By day 35 post-anastomosis, the number of prosaposin-positive neurons in the operated side recovered to normal levels. The number of prosaposin-positive neurons, as well as immunoreactivity intensity, was significantly greater in the facial nerve nucleus, compared with the transection group on days 14, 21, and 35 post-surgery (P < 0.05). The rhythmic whisking of vibrissa recovered, and recovery time was consistent with increased numbers of prosaposin-positive neurons.CONCLUSION Within 7 days after injury, prosaposin expression in the facial nerve nucleus exhibited an initial increase, followed by a decrease, and was not affected by facial nerve repair. Following facial nerve damage, neural anastomosis was shown to increase prosaposin expression in the facial nerve nucleus after 14 days. Recovery of prosaposin occurred simultaneously with reinnervation.更多还原