面神经切除后运动皮层面区与合谷穴区神经投射研究

发布时间：2018-05-27 22:20

本文选题：功能重组 + 结构基础　；参考：《广州中医药大学》2017年博士论文

【摘要】：目的:通过切除面神经干,运用运动皮层皮层内电刺激和神经示踪方法,研究大鼠和恒河猴面区与合谷穴区的功能重组及其结构基础,探讨病理状态下"面口合谷收"的经脉特异性联系的生物学机制。方法:首先为了摸索面区传出通路神经投射情况和观察精细神经结构的最佳条件,先对神经示踪技术在面区传出通路的应用作前期研究。选取17只成年雄性大鼠为研究对象,在丘脑腹后内侧核压力注射高分子量生物素化葡聚糖胺,观察丘脑至皮层,皮层至丘脑通路顺行和逆行的示踪效果,并探讨其示踪效果与注射后动物存活时长的关系,从而了解其神经示踪特性。然后把神经示踪技术运用在大鼠实验。选取10只大鼠,5只大鼠为模型组,5只为空白对照组。予模型组大鼠面神经干切除,20至30天后,进行运动皮层皮层内电刺激,绘制大鼠运动皮层躯体定位图,并与对照组作对比分析。皮层内电刺激结束后,分别在运动皮层面区注射生物素化葡聚糖胺,在运动皮层手区注射霍乱毒素B亚单位。7天后予灌流取脑,进行形态学研究,观察运动皮层面区手区神经元、神经纤维的标记情况及皮层之间的投射情况,比较两组单位面积内单根轴突上膨体数。在前两项实验的研究基础上,最后在恒河猴上进行实验。选取4只恒河猴,两只为模型组,两只为空白对照组。予模型组两只恒河猴进行面神经干切除,一只造模时长为一年,一只造模时长为1个月。分别对四只恒河猴进行运动皮层皮层内电刺激,绘制恒河猴运动皮层躯体定位图。皮层内电刺激结束后,分别在运动皮层面区注射生物素化葡聚糖胺,在运动皮层合谷穴区注射霍乱毒素B亚单位。7至10天后灌流取脑和脊髓进行形态学研究。主要观察运动皮层面区与合谷穴区、脊髓的神经元、神经纤维的标记情况及面神经核中神经元与星形胶质细胞的形态。结果:1.研究发现高分子量生物素化葡聚糖胺是一种灵敏的双向示踪剂,在最佳的存活期可以展现出精细的神经元、树突、轴突的形态。高分子量生物素化葡聚糖胺在丘脑-皮层投射通路和皮层-丘脑投射通路的最佳存活期是十天。另外其结合荧光染料链霉亲和素-Alexa Fluor 594在激光扫描共聚焦显微镜下可以展现细致清楚的神经结构,比ABC染色法好。2.大鼠面神经干切除后,进行皮层内电刺激,两组总记录点数差异无统计学意义(P0.05),两组手区记录点在躯体定位图X轴分布差异有统计学意义(P0.05),手区向面区扩展。比较两组轴突单位面积内单根轴突上膨体数,P0.05,差异有统计学意义,模型组轴突上的膨体密度比对照组少。运动皮层与同侧感觉皮层、对侧运动皮层、对侧感觉皮层存在相互投射关系。3.恒河猴面神经干切除后,进行沿中央沟运动皮层皮层内电刺激,发现合谷穴区向面区扩展。面神经切除1月恒河猴运动皮层膨体稀疏。通过观察面神经核神经元和星形胶质细胞,发现切除面神经干后患侧面神经核神经元出现病理变化,星形胶质细胞增多,考虑造模后患侧面神经核发生退行性病变。结论:面神经干切除后,大鼠和恒河猴运动皮层合谷穴区向面区扩展,说明运动皮层面区合谷穴区发生功能重组。面区轴突的膨体密度减小,推测为运动皮层面区功能重组的结构基础。这可能是病理状态下"面口合谷收"的生物学机制之一。
[Abstract]:Objective: To study the functional reorganization and structural basis of the rat and Ganges RIver monkey area and Hegu area by excising the facial nerve trunk and using the cortical electrical stimulation and the nerve tracing method in the cortex of the motor cortex, and to explore the biological mechanism of the specific relationship between the meridian and the valley in the pathological state. The best conditions for the projection and observation of the fine nerve structure were first studied by the application of the neural tracer technique in the surface area. 17 adult male rats were selected as the research object. The pressure of the ventral posteromedial nucleus of the thalamus was injected with high molecular weight biotinylated dextran amines, and the thalamus to cortex, cortex to thalamus pathway were observed and reversed. The tracer effect and the relationship between the tracer effect and the survival length of the animals after the injection were discussed, and the neural tracer characteristics were understood. Then the neural tracer technique was used in rats. 10 rats were selected, 5 rats were used as model groups and 5 were blank control groups. The facial nerve was excised in the model group, and the motor cortex was performed 20 to 30 days later. Electric stimulation in cortex was used to plot the somatic location of the motor cortex in rats, and compared with the control group. After the end of the cortical electric stimulation, the biotinylated dextran amine was injected into the motor cortex area, and the subunit of cholera toxin B was injected into the brain in the hand area of the motor cortex for.7 days, and the morphological study was carried out to observe the hand of the skin layer area. On the basis of the first two experiments on the Ganges RIver monkey, 4 Ganges RIver monkeys were selected, two were model groups and two were empty white control groups, and two Ganges RIver monkeys in the model group were given face God. After excision, one model was one year long and a model was 1 months long. Four Ganges RIver monkeys were stimulated by electric cortical stimulation in the motor cortex to map the physical location of the motor cortex of Ganges RIver monkey. After the electrical stimulation in the cortex, the biotinylated dextran amine was injected into the motor cortex area and cholera was injected into the Hegu area of the motor cortex. The brain and spinal cord of the toxin B subunit were injected into the brain and the spinal cord for morphological study after 10 days. The primary observation of the motor cortex area and Hegu area, the neurons of the spinal cord, the labeling of the nerve fibers and the morphology of the neurons in the nucleus and astrocytes in the nucleus of the facial nerve. Results: 1. the results showed that the high score of biotinylated glucan amine was a sensitive one. Bi-directional tracers, in the best survival period, can show fine neurons, dendrites, and axons. The optimal survival time of high molecular weight biotinylated glucan amine in the thalamocortical projection pathway and cortical thalamus projection pathway is ten days. In addition, it combines with fluorescent dye streptomycin -Alexa Fluor 594 in laser scanning confocal microscopy Microscopically, a detailed and clear neural structure could be displayed, and there was no significant difference between the two groups of the total records in the total number of records in the two groups (P0.05). The difference in the distribution of the X axis in the two groups of hand areas was statistically significant (P0.05), and the two groups of axons were compared. The number of bulge in the single axon per unit area, P0.05, the difference was statistically significant. The swelling density on the axon of the model group was less than that of the control group. There was a mutual projection relationship between the motor cortex and the ipsilateral sensory cortex, the lateral motor cortex and the contralateral sensory cortex,.3. Ganges RIver monkey facial nerve was excised, and the electric stimulation along the cortex of the central groove motor cortex was carried out. The expansion of the area of the Hegu area was found. In January, the cortical swelling of the motor cortex of Ganges RIver monkey was sparse. By observing the neurons of the facial nerve and astrocytes, the pathological changes of the neurons in the lateral nerve nucleus after the removal of the facial nerve were found, and the astrocytes were increased. The degenerative lesion of the lateral nerve nucleus after the model was considered. Theory: after the facial nerve trunk excision, the rats and the Hegu acupoint area of the motor cortex of the Ganges RIver monkey expand to the surface area, indicating the functional reorganization of the Hegu area in the motor cortex area. The expansion density of the surface axon decreases, which is presumed to be the structural basis for the functional reorganization of the motor cortex area. This may be one of the biological mechanisms of the "facial joint Valley harvest" under the pathological condition.
【学位授予单位】：广州中医药大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R245

【参考文献】