本文選題：功能重組 + 結構基礎　； 參考：《廣州中醫(yī)藥大學》2017年博士論文

【摘要】：目的:通過切除面神經干,運用運動皮層皮層內電刺激和神經示蹤方法,研究大鼠和恒河猴面區(qū)與合谷穴區(qū)的功能重組及其結構基礎,探討病理狀態(tài)下"面口合谷收"的經脈特異性聯(lián)系的生物學機制。方法:首先為了摸索面區(qū)傳出通路神經投射情況和觀察精細神經結構的最佳條件,先對神經示蹤技術在面區(qū)傳出通路的應用作前期研究。選取17只成年雄性大鼠為研究對象,在丘腦腹后內側核壓力注射高分子量生物素化葡聚糖胺,觀察丘腦至皮層,皮層至丘腦通路順行和逆行的示蹤效果,并探討其示蹤效果與注射后動物存活時長的關系,從而了解其神經示蹤特性。然后把神經示蹤技術運用在大鼠實驗。選取10只大鼠,5只大鼠為模型組,5只為空白對照組。予模型組大鼠面神經干切除,20至30天后,進行運動皮層皮層內電刺激,繪制大鼠運動皮層軀體定位圖,并與對照組作對比分析。皮層內電刺激結束后,分別在運動皮層面區(qū)注射生物素化葡聚糖胺,在運動皮層手區(qū)注射霍亂毒素B亞單位。7天后予灌流取腦,進行形態(tài)學研究,觀察運動皮層面區(qū)手區(qū)神經元、神經纖維的標記情況及皮層之間的投射情況,比較兩組單位面積內單根軸突上膨體數(shù)。在前兩項實驗的研究基礎上,最后在恒河猴上進行實驗。選取4只恒河猴,兩只為模型組,兩只為空白對照組。予模型組兩只恒河猴進行面神經干切除,一只造模時長為一年,一只造模時長為1個月。分別對四只恒河猴進行運動皮層皮層內電刺激,繪制恒河猴運動皮層軀體定位圖。皮層內電刺激結束后,分別在運動皮層面區(qū)注射生物素化葡聚糖胺,在運動皮層合谷穴區(qū)注射霍亂毒素B亞單位。7至10天后灌流取腦和脊髓進行形態(tài)學研究。主要觀察運動皮層面區(qū)與合谷穴區(qū)、脊髓的神經元、神經纖維的標記情況及面神經核中神經元與星形膠質細胞的形態(tài)。結果:1.研究發(fā)現(xiàn)高分子量生物素化葡聚糖胺是一種靈敏的雙向示蹤劑,在最佳的存活期可以展現(xiàn)出精細的神經元、樹突、軸突的形態(tài)。高分子量生物素化葡聚糖胺在丘腦-皮層投射通路和皮層-丘腦投射通路的最佳存活期是十天。另外其結合熒光染料鏈霉親和素-Alexa Fluor 594在激光掃描共聚焦顯微鏡下可以展現(xiàn)細致清楚的神經結構,比ABC染色法好。2.大鼠面神經干切除后,進行皮層內電刺激,兩組總記錄點數(shù)差異無統(tǒng)計學意義(P0.05),兩組手區(qū)記錄點在軀體定位圖X軸分布差異有統(tǒng)計學意義(P0.05),手區(qū)向面區(qū)擴展。比較兩組軸突單位面積內單根軸突上膨體數(shù),P0.05,差異有統(tǒng)計學意義,模型組軸突上的膨體密度比對照組少。運動皮層與同側感覺皮層、對側運動皮層、對側感覺皮層存在相互投射關系。3.恒河猴面神經干切除后,進行沿中央溝運動皮層皮層內電刺激,發(fā)現(xiàn)合谷穴區(qū)向面區(qū)擴展。面神經切除1月恒河猴運動皮層膨體稀疏。通過觀察面神經核神經元和星形膠質細胞,發(fā)現(xiàn)切除面神經干后患側面神經核神經元出現(xiàn)病理變化,星形膠質細胞增多,考慮造模后患側面神經核發(fā)生退行性病變。結論:面神經干切除后,大鼠和恒河猴運動皮層合谷穴區(qū)向面區(qū)擴展,說明運動皮層面區(qū)合谷穴區(qū)發(fā)生功能重組。面區(qū)軸突的膨體密度減小,推測為運動皮層面區(qū)功能重組的結構基礎。這可能是病理狀態(tài)下"面口合谷收"的生物學機制之一。
[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.
【學位授予單位】：廣州中醫(yī)藥大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R245

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