UL对清醒大鼠MVN区Glu含量和c-Fos蛋白表达的影响

发布时间：2018-01-17 18:00

本文关键词：UL对清醒大鼠MVN区Glu含量和c-Fos蛋白表达的影响　出处：《延边大学》2017年硕士论文　论文类型：学位论文

【摘要】：前庭器官,可以感受空间位置和运动状态。末梢前庭感受器将躯体的运动和头部空间位置转变为传入信号,经前庭神经纤维通路传至前庭神经核(vestibular nucleus,VN)。VN接受外周前庭器官的神经信号,并与视觉和本体感觉共同维持机体平衡状态。单侧前庭迷路破坏术(unilaterallabyrinthectomy,UL)后,前庭代偿是研究病变后中枢神经系统可塑性的理想实验模型。UL后,可出现头偏位、眼震颤和水平旋转等体征。但是前庭功能失衡体征和症状会随着时间的推移逐渐消失,称为前庭代偿(vestibularcompensation,VC)。韩国的朴炳林等人在破坏外周前庭器官的麻醉大鼠观察到,前庭代偿急性期双侧前庭神经内侧核区c-Fos不对称表达,大约6 d这种不对称表达消失。本实验室发现,麻醉大鼠单侧耳内灌注2%利多卡因之后,短时阻断了前庭信号的传入,前庭内侧核(medial vestibular neuclei,MVN)谷氨酸(glutamate,Glu)的含量出现不对称表达,说明Glu可能与前庭代偿有关联。但是,到目前为止这些实验都是在麻醉动物进行的,尚无前庭代偿不同时期MVN区谷氨酸含量变化的系统研究报道,更无在清醒状态下的相关研究。本研究拟在清醒动物利用核团微量透析、高效液相分析、免疫组织化学和蛋白印迹法,观察单侧破坏外周前庭器官不同时期清醒大鼠MVN区Glu含量和c-Fos蛋白动态变化,探讨前庭代偿过程中是有Glu及其NMDA受体的参与。实验结果如下:1.UL后,6h头偏向损伤侧,并在24h开始加重,72h达到高峰并不缓解。眼震颤在破坏24 h达到最高峰,48 h迅速缓解,破坏120 h消失。水平旋转在破坏6h出现12h后减弱,96h达到高峰,168 h消失。2.UL后,MVN区细胞外液中Glu的含量:损伤同侧24、12 h达高峰,144 h基本恢复;损伤对侧6 h达高峰,12小时下降,48 h接着增高,72 h达第二次高峰,144 h基本恢复。MVN区双侧在6、12、24、72、96 h、存在着显著差异(p0.05,n=12)。3.UL后,MVN区中c-Fos免疫阳性神经元的数量:损伤同侧24,12 h达高峰,96 h基本恢复;损伤对侧6 h达高峰,12小时下降,48 h接着增高,72 h达第二次高峰。MVN区双侧在6、12、24、48、72、96、120 h存在着显著差异(p0.05,n=6)。MVN区144h后c-Fos免疫阳性神经元基本消失。4.UL后,MVN中c-Fos蛋白的表达:损伤同侧24,12h达高峰,144h基本消失;损伤6 h后不对称表达明显高于损伤同侧,12小时下降,48 h接着增高,72 h达第二次高峰。MVN区双侧在6、12、24、48、72、96 h、120 h存在着显著差异(p0.05,n=6)。MVN区144h后c-Fos蛋白基本消失。5.UL 72 h侧脑室注射MK-801后,MVN内c-Fos蛋白的表达具有显著性差异(p0.05,n=6)。以上结果提示:1.清醒大鼠前庭代偿过程中可能有Glu的参与;2.清醒大鼠前庭代偿过程中会引起c-Fos蛋白的表达变化;3.清醒大鼠前庭代偿过程Glu可能通过NMDA受体介导神经信号的传递。
[Abstract]:The vestibular organ can feel the spatial position and the motion state. The vestibular receptor changes the body movement and the head space position into the afferent signal. The vestibular nerve fibers were transmitted to the vestibular nucleus vestibular nucleus via the vestibular nerve fiber pathway to receive the nerve signals from the vestibular organs. And together with vision and proprioceptive sensation to maintain the balance of the body. Unilateral vestibular labyrinth destruction after unilateral abyrinthe tomyULL. Vestibular compensation is an ideal experimental model to study the plasticity of central nervous system after pathological changes. Eye tremor and horizontal rotation. But vestibular dysfunction signs and symptoms will gradually disappear with the passage of time, called vestibular compensation. Park Bing-lin of Korea observed asymmetric expression of c-Fos in bilateral medial vestibular nucleus during the acute phase of vestibular compensation in anesthetized rats which destroyed peripheral vestibular organs. The asymmetric expression disappeared at about 6 days. We found that the afferent of vestibular signal was blocked shortly after 2% lidocaine was infused into the single lateral ear of anesthetized rats. There was asymmetric expression of glutamate in medial vestibular neuclei of the medial vestibular nucleus. These results suggest that Glu may be associated with vestibular compensation. However, so far, these experiments have been carried out in anesthetized animals, and there is no systematic research report on the changes of glutamate content in MVN region at different stages of vestibular compensation. This study was designed to use micronucleus dialysis, high performance liquid analysis, immunohistochemistry and Western blotting in awake animals. To observe the dynamic changes of Glu content and c-Fos protein in MVN region of conscious rats in different periods of unilateral destruction of peripheral vestibular organs. Glu and its NMDA receptors were involved in the process of vestibular compensation. The results were as follows: 1. After 1. 1. The head tilted to the injured side 6 hours after UL, and began to increase at 24 hours. Reaching the peak at 72 h did not relieve, but the eye tremor reached the peak at 24 h and reached the peak at 48 h, and disappeared at 120 h, and the horizontal rotation weakened after 6 h damage and 12 h. The content of Glu in the extracellular fluid of MVN region disappeared at 96 h and disappeared at 168h. 2. The content of Glu in the extracellular fluid of MVN region: the ipsilateral side reached the peak at 24 h and reached the peak at 144h. The contralateral side reached the peak at 6 h, reached the peak at 12 h and decreased at 48 h, then increased at 72 h and reached the second peak at 144 h. At 96 h, there was a significant difference in the number of c-Fos immunoreactive neurons in the MVN region after p0.05. 3. The number of c-Fos immunoreactive neurons in the MVN region reached its peak at 24 h after injury. 96 h recovery; The contralateral side reached the peak at 6 h and decreased at 12 h and decreased at 48 h, then increased to the second peak at 72 h. There was a significant difference at 120 h after the disappearance of c-Fos immunoreactive neurons. The expression of c-Fos protein in MVN: the ipsilateral injury reached the peak at 24 h and reached the peak at 14 4 h. The asymmetrical expression of MVN was significantly higher than that of ipsilateral injury at 12 hours and 48 hours after injury, and then increased to the second peak at 72 hours. There was significant difference between 96 h and 120 h (p 0.05). The c-Fos protein disappeared basically at 144h after 72 h injection of MK-801 into the lateral ventricle. There was significant difference in the expression of c-Fos protein in MVN. The above results suggested that Glu might be involved in the vestibular compensatory process of conscious rats. 2.The expression of c-Fos protein was changed during the process of vestibular compensation in conscious rats. 3. During the vestibular compensatory process, Glu may mediate neural signal transduction by NMDA receptor.
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R33

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