針刺任督脈經(jīng)穴對(duì)腦干吞咽中間神經(jīng)元的調(diào)節(jié)作用
本文選題:針刺 + 吞咽; 參考:《廣州中醫(yī)藥大學(xué)》2015年博士論文
【摘要】:吞咽是人類最為基本和重要的生理功能之一,健康人每天進(jìn)行無意識(shí)的吞咽活動(dòng)可達(dá)一千次,其主要作用是將食物由口咽部傳遞至食管。其整個(gè)過程需要6對(duì)顱神經(jīng)和25塊肌肉協(xié)同完成。吞咽是一個(gè)復(fù)雜的感覺運(yùn)動(dòng)活動(dòng),依靠大腦皮層、腦干吞咽中樞和顱神經(jīng)(Ⅴ、Ⅸ、Ⅹ、(?) (?))之間高度系統(tǒng)性的相互作用。這一過程既有主動(dòng)成分又有非主動(dòng)成分,分別反應(yīng)了皮層和腦干的中樞調(diào)節(jié)途徑。針刺作為一種在臨床上治療吞咽障礙、調(diào)節(jié)吞咽功能行之有效的方法,其內(nèi)在機(jī)理一直未有明確的認(rèn)識(shí),近20年來,圍繞吞咽反射的腦干神經(jīng)元網(wǎng)絡(luò)所開展的大量研究使得我們對(duì)吞咽的生理學(xué)機(jī)制有了一個(gè)較為清晰的認(rèn)識(shí),腦干吞咽神經(jīng)元網(wǎng)絡(luò)主要由傳入纖維、中間神經(jīng)元和運(yùn)動(dòng)神經(jīng)元組成。針刺作用在吞咽腦干神經(jīng)元網(wǎng)絡(luò)的機(jī)制尚未清楚,這些研究的基礎(chǔ)為我們研究針刺調(diào)節(jié)吞咽功能提供了一定的思路,因此,本研究針對(duì)腦干背側(cè)吞咽中樞,探討針刺對(duì)該吞咽初級(jí)中樞的作用機(jī)制。目的:觀察針刺對(duì)腦干背側(cè)吞咽中間神經(jīng)元的激活情況,從生理狀況下神經(jīng)電生理情況、損毀核團(tuán)后針刺效應(yīng)變化以及針刺對(duì)神經(jīng)解剖形態(tài)學(xué)的效應(yīng)幾個(gè)方面,來探討針刺對(duì)初級(jí)中樞吞咽中間神經(jīng)元的作用,揭示針刺調(diào)節(jié)吞咽功能的可能機(jī)理。實(shí)驗(yàn)一針刺對(duì)吞咽相關(guān)中間神經(jīng)元(孤束核)放電的調(diào)節(jié)方法:SD大鼠55只,雌雄不限,水合氯醛麻醉后,先進(jìn)行前頸部手術(shù),暴露下頜舌骨肌以及喉上神經(jīng),分別連接記錄電極以及刺激電極,再進(jìn)行腦部手術(shù),顱骨鉆孔后暴露延髓閂部,用立體定位儀將玻璃微電極插入延髓孤束核內(nèi),開始記錄神經(jīng)元放電,先刺激喉上神經(jīng)引發(fā)肌電變化及吞咽,確定吞咽相關(guān)神經(jīng)元后,分別針刺風(fēng)府穴、廉泉穴、內(nèi)關(guān)穴以及足三里穴30s,觀察神經(jīng)元放電變化以及肌電變化。記錄完畢后標(biāo)記記錄位置,進(jìn)行組織學(xué)鑒別,定位不準(zhǔn)確的數(shù)據(jù)棄之不用。結(jié)果:本實(shí)驗(yàn)共記錄到時(shí)相型吞咽相關(guān)神經(jīng)元共63個(gè),對(duì)針刺“風(fēng)府”穴起反應(yīng)的神經(jīng)元共41個(gè)(65%),“廉泉”穴36個(gè)(57%),“內(nèi)關(guān)”穴23個(gè)(37%),“足三里”穴19個(gè)(30%)。自發(fā)放電型神經(jīng)元共完整記錄到107個(gè),與吞咽運(yùn)動(dòng)相關(guān)的神經(jīng)元共記錄到70個(gè),主要為表現(xiàn)為興奮性神經(jīng)元。針刺“風(fēng)府”穴后,NTS神經(jīng)元放電頻率增加的共49個(gè),減少的共6個(gè),無變化的共15個(gè);針刺“廉泉”后NTS神經(jīng)元放電頻率增加的共53個(gè),減少的共5個(gè),無變化的共12個(gè),針刺“內(nèi)關(guān)”后NTS神經(jīng)元放電頻率增加的共38個(gè),減少的共3個(gè),無變化的共29個(gè),針刺足三里后,NTS神經(jīng)元放電頻率增加的共31個(gè),減少的共11個(gè),無變化的共28個(gè)。針刺以上四穴對(duì)孤束核吞咽相關(guān)神經(jīng)元的激活作用,以廉泉穴及風(fēng)府穴較為顯著,內(nèi)關(guān)與足三里對(duì)吞咽相關(guān)孤束核神經(jīng)元均有-定程度的激活作用,其中對(duì)各組興奮性神經(jīng)元放電頻率變化率進(jìn)行比較,針刺廉泉穴及風(fēng)府穴與針刺其他穴位分別比較,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論:針刺后可誘發(fā)吞咽相關(guān)神經(jīng)元的放電變化,表明針刺對(duì)腦干背側(cè)吞咽中樞神經(jīng)元具有激活作用,在穴位的選擇上,廉泉穴與風(fēng)府穴對(duì)神經(jīng)元激活作用明顯,內(nèi)關(guān)穴及足三里穴也可一定程度的激活該神經(jīng)元。共12個(gè),針刺“內(nèi)關(guān)”后NTS神經(jīng)元放電頻率增加的共38個(gè),減少的共3個(gè),無變化的共29個(gè),針刺足三里后,NTS神經(jīng)元放電頻率增加的共31個(gè),減少的共11個(gè),無變化的共28個(gè)。針刺以上四穴對(duì)孤束核吞咽相關(guān)神經(jīng)元的激活作用,以廉泉穴及風(fēng)府穴較為顯著,內(nèi)關(guān)與足三里對(duì)吞咽相關(guān)孤束核神經(jīng)元均有-定程度的激活作用,其中對(duì)各組興奮性神經(jīng)元放電頻率變化率進(jìn)行比較,針刺廉泉穴及風(fēng)府穴與針刺其他穴位分別比較,差異具有統(tǒng)計(jì)學(xué)意義(P0.05)。實(shí)驗(yàn)二 損毀孤束核對(duì)針刺任督脈經(jīng)穴調(diào)節(jié)吞咽肌運(yùn)動(dòng)的影響方法:SD大鼠42只,分為風(fēng)府組、廉泉組以及內(nèi)關(guān)組,每組均分為損毀和假損毀兩部分,麻醉后進(jìn)行頸部手術(shù)同實(shí)驗(yàn)一,僅放置肌電記錄電極,腦部手術(shù)與實(shí)驗(yàn)一相同,先對(duì)各組進(jìn)行針刺60s,觀察下頜舌骨肌肌電放電及吞咽頻數(shù),然后在立體定位儀引導(dǎo)下將金屬電極插入孤束核相關(guān)核團(tuán),通陽極直流電0.1mA,時(shí)間20s,進(jìn)行核團(tuán)電損毀。損毀結(jié)束后再次針刺以上三組穴位,觀察肌電變化以及吞咽頻數(shù)。假損毀實(shí)驗(yàn)中,僅將電極插入核團(tuán),不進(jìn)行通電,時(shí)間也為20s。實(shí)驗(yàn)結(jié)束后,對(duì)延髓進(jìn)行組織學(xué)檢測(cè),損毀部位不準(zhǔn)確者數(shù)據(jù)棄之不用。結(jié)果:在損毀孤束核前,針刺“廉泉穴”組最多引發(fā)的下頜舌骨肌放電頻數(shù),較風(fēng)府組與內(nèi)關(guān)組均有顯著差異(P0.05),而針刺“風(fēng)府”穴與“內(nèi)關(guān)”穴兩組比較無明顯差異(P0.05)。損毀前后進(jìn)行針刺,以上三組進(jìn)行損毀前后組內(nèi)比較,損毀后針刺引發(fā)的下頜舌骨肌肌電放電頻次較損毀前明顯減少,其中以廉泉穴組以及風(fēng)府穴組變化最為明顯,損毀前后具有明顯差異(P0.01),針刺內(nèi)關(guān)穴在損毀前后吞咽頻次差異也具有統(tǒng)計(jì)學(xué)意義(P0.05)。損毀孤束核后對(duì)三組進(jìn)行組間比較,針刺不同穴位對(duì)吞咽的影響比較無明顯差別(P0.05)。假損毀對(duì)照試驗(yàn)顯示,針刺風(fēng)府穴、廉泉穴以及內(nèi)關(guān)穴,在假損毀前后誘發(fā)的吞咽頻數(shù)無明顯差異結(jié)論:電解損毀孤束核后,針刺“廉泉”、“風(fēng)府”以及“內(nèi)關(guān)”穴對(duì)大鼠的吞咽誘發(fā)作用明顯減弱,提示孤束核是針刺調(diào)節(jié)吞咽功能中的一個(gè)重要的初級(jí)中樞。針刺廉泉穴對(duì)吞咽的誘發(fā)作用明顯,風(fēng)府穴及內(nèi)關(guān)穴也可在一定程度上誘發(fā)吞咽運(yùn)動(dòng),提示穴位的選擇對(duì)針刺調(diào)節(jié)吞咽功能具有一定的意義。實(shí)驗(yàn)三針刺對(duì)孤束核中c-fos蛋白表達(dá)的影響方法:SD大鼠24只,分為空白組、風(fēng)府組、廉泉組以及內(nèi)關(guān)組,常規(guī)麻醉后,進(jìn)行頸部手術(shù),暴露下頜舌骨肌后插入記錄電極,分別電針上述三組穴位,頻率10Hz,2mA,共20min,同時(shí)記錄下頜舌骨肌肌電變化以及吞咽運(yùn)動(dòng),針刺完畢后,對(duì)大鼠進(jìn)行灌流取腦,保留延髓及部分脊髓,固定脫水后進(jìn)行冰凍切片,并用免疫組化方法觀察大鼠延髓相應(yīng)區(qū)域的c-fos蛋白表達(dá)變化。結(jié)果:電針各組大鼠誘發(fā)吞咽肌放電以針刺廉泉穴及風(fēng)府穴較為顯著,效果由于內(nèi)關(guān)組(P0.05),空白組則無吞咽運(yùn)動(dòng)產(chǎn)生。針刺后大鼠各組c-fos蛋白陽性表達(dá)方面,在孤束核區(qū)域范圍內(nèi),空白組在孤束核區(qū)域內(nèi)可見零星的c-fos陽性表達(dá)細(xì)胞,其余三組均可見陽性表達(dá)的細(xì)胞,其中以廉泉穴組、風(fēng)府組以及內(nèi)關(guān)組均有較多的陽性表達(dá)細(xì)胞表達(dá),三組比較無顯著差異。結(jié)論:本實(shí)驗(yàn)證實(shí)了針刺廉泉穴、風(fēng)府穴不僅可以直接誘發(fā)吞咽肌放電變化,還可使孤束核內(nèi)c-fos蛋白表達(dá)升高,從形態(tài)學(xué)的角度進(jìn)一步表明了孤束核參與了針刺調(diào)節(jié)吞咽運(yùn)動(dòng)并起到重要的作用。
[Abstract]:Swallowing is one of the most basic and important physiological functions of human beings. A healthy person carries out an unconscious swallowing activity one thousand times a day. Its main function is to transfer food from the mouth to the esophagus. The whole process requires 6 pairs of cranial nerves and 25 muscles. Swallowing is a complex sensory exercise, depending on the cerebral cortex. A highly systematic interaction between the brain stem swallowing center and the cranial nerve (V, IX,?). This process has both active and non active components, reflecting the central regulatory pathways of the cortex and brain stem respectively. Acupuncture is an effective method for the clinical treatment of swallowing disorders and regulation of swallowing function. The internal mechanism is one of the mechanisms. There is no clear understanding. In the past 20 years, a large number of studies on the brain stem neuron network around swallowing reflex have made a clearer understanding of the physiological mechanism of swallowing. The brain stem swallowing neuron network is mainly composed of afferent fibers, intermediate neurons and transport neurons. The action of acupuncture on the swallowing brainstem nerve The mechanism of the meta network is not yet clear. The basis of these studies provides a certain way of thinking on the regulation of the swallowing function by acupuncture. Therefore, this study aims at the mechanism of acupuncture on the primary center of the swallowing in the dorsal swallowing center of the brain stem. Objective: To observe the activation of the intermediate neurons in the dorsum of the brain stem by acupuncture. Under the condition of neurophysiology, the changes in the acupuncture effect and the effect of acupuncture on the anatomic morphology of the nerve after the destruction of the nucleus, the effect of acupuncture on the intermediate neurons of the primary central swallowing is discussed, and the possible mechanism of acupuncture to regulate the swallowing function is revealed. Method: 55 SD rats, male and female, and male and female, after chloral hydrate anaesthetized, first undergo anterior cervical surgery, expose the mandibular hyoid muscle and the superior laryngeal nerve, connect recording electrodes and stimulation electrodes respectively, then perform brain surgery, expose the latch of the medulla after the skull drilling, and insert the glass microelectrode into the nucleus of the medulla of the medulla of the medulla of the medulla and start recording with a stereoscopic instrument. Neurons discharge, first stimulation of the upper laryngeal nerve to induce electromyographic changes and swallowing, and determine the dysphagia related neurons, then acupuncture at the Feng Fu point, the Lian Quan point, the Neiguan point and the Zusanli point 30s, to observe the changes in the neuron discharge and the electromyography. Results: a total of 63 dysphagia related neurons were recorded in the experiment. There were 41 (65%) neurons responding to the acupuncture "Feng Fu" point, 36 (57%) "Lian Quan" point, 23 (37%) and 19 (30%) points of "Zusanli" acupoint. A total of 107 neurons were recorded in a total number of neurons associated with swallowing movement. 70 of them were recorded as excitatory neurons. After acupuncture of "Feng Fu" point, the discharge frequency of NTS neurons increased in a total of 49, a total of 6 decreased, and no change in total, and there were 53 increases in the discharge frequency of NTS neurons after the needling of "Lian Quan", a total of 5 reduced, 12 without changes, and the discharge frequency of NTS neurons after needling "Neiguan". A total of 38 increased rates, a total of 3 decreased, and no change in total 29. After the acupuncture of Zusanli, the discharge frequency of NTS neurons increased in a total of 31, reduced in a total of 11, without a total of 28. The activation of the four points above the four acupoints to the nucleus of the nucleus of the solitary tract was more significant in the Lian Quan point and the Feng Fu acupoint, and the Neiguan and Zusanli on the swallowing related soliton The activation of the nucleus neurons in the nucleus had a degree of activation, in which the frequency of the discharge frequency of the excitatory neurons in each group was compared. The difference was statistically significant (P0.05). Conclusion: after acupuncture, the changes in the discharge of the related neurons of the swallowing were induced, indicating that acupuncture on the back of the brain stem. In the selection of the acupoint, the activation of the neurons of the Lian Quan point and the Feng Fu point is obvious in the selection of the acupoints, and the Neiguan point and the Zusanli point can also activate the neuron to a certain extent. There are altogether 38 increases in the discharge frequency of NTS neurons after the needling of "Neiguan", and a total of 3 decreased, without change of 29. After Zusanli, the discharge frequency of NTS neurons increased in a total of 31, a total of 11 decreased, and no change in the total 28. The activation of the four points of acupuncture above to the nucleus of the nucleus of the solitary tract was more significant in the Lian Quan and Feng Fu points, and the activation of the Neiguan and Zusanli on the deglutition related nucleus of the nucleus of the deglutition related solitary tract was activated. Comparison of the frequency change rate of the discharge of the excitatory neurons, the difference was statistically significant (P0.05). The difference was statistically significant (P0.05). In experiment, the effect of the two damaged solitary tract on the regulation of the movement of the swallowing muscle by acupuncture at the meridian point of the meridian was divided into 42 rats, which were divided into the Feng Fu Group, the Lian Quan Group and the Neiguan group. Two parts were divided into damage and false damage. After the anaesthesia, the neck surgery was performed with the experiment one. Only the electromyographic recording electrode was placed, the brain operation was the same as the experimental one. First, the acupuncture 60s was used to observe the myoelectric discharge and swallowing frequency of the mandibular hyoid muscle. Then the metal electrode was inserted into the nucleus of the nucleus of the solitary tract under the guidance of the stereotaxis. 0.1mA, time 20s, the time of nuclear damage. After the damage, the above three groups of acupoints were needled again to observe the changes of electromyography and swallowing frequency. In the false damage experiment, only the electrode was inserted into the nucleus and no electricity was carried out. The time is also the tissue test of the medulla after the end of the 20s. experiment. Before the damage of the nucleus of the solitary tract, the frequency of the discharge of the mandibular hyoid muscle in the acupuncture "Lian Quan point" group was significantly different (P0.05), but there was no significant difference (P0.05) between the two groups of the "Feng Fu" point and the "Neiguan" point (P0.05). The frequency of electroelectric stimulation of the mandibular hyoid muscle was significantly reduced before the damage. The changes were most obvious in the Lian Quan point group and the Feng Fu point group, and there were significant differences before and after the damage (P0.01). The difference of the swallowing frequency of the acupuncture Neiguan point before and after the damage was also statistically significant (P0.05). The comparison of the three groups after the damage of the nucleus of the solitary tract was compared. There was no significant difference in the effect of different acupoints on swallowing (P0.05). The false damage control test showed that there was no obvious difference in swallowing frequency induced by acupuncture at the Feng Fu point, Lian Quan point and Neiguan point before and after the false damage: after the electrolytic destruction of the nucleus of the solitary tract, the stimulation of the "Lian Quan", "Feng Fu" and "Neiguan" on the swallowing induced effect on rats It is obvious that the nucleus of solitary tract is an important primary center in regulating the swallowing function of the needling. The effect of acupuncture at Lian Quan point on swallowing is obvious, and the Feng Fu point and Neiguan point can also induce swallowing movement to a certain extent, suggesting that the selection of acupoints has certain significance on the regulation of the swallowing function by acupuncture. Experiment three acupuncture on the nucleus of solitary tract in the nucleus of solitary tract. The influence methods of c-fos protein expression: 24 SD rats were divided into blank group, Feng Fu Group, Lian Quan Group and Neiguan group. After routine anesthesia, cervical surgery was performed, and the mandibular hyoid muscle was exposed to record electrodes. The above three groups of acupoints were electroacupuncture respectively, the frequency 10Hz, 2mA, 20min, and the recording of the myoelectric changes of the mandibular hyoid muscle and the swallowing movement and acupuncture. After completion, the rats were injected into the brain, retained the medulla oblongata and part of the spinal cord, fixed the frozen section after dehydration, and observed the changes in the expression of c-fos protein in the corresponding region of the medulla oblongata in the rats by immunohistochemistry. Results: the rats of the electroacupuncture group induced the deglutition of the swallowing muscle to be more significant in the needling of the Lian Quan point and the Feng Fu point. The effect was due to the Neiguan group (P0.05 In the blank group, there was no swallowing movement. The positive expression of c-fos protein in the rats after acupuncture was found in the region of the nucleus of the solitary tract. The blank group showed sporadic c-fos positive cells in the nucleus of the nucleus of the solitary tract, and the other three groups were all positive cells, among which there were more positive forms in the Lian Quan point group, the Feng Fu group and the Neiguan group. There was no significant difference between the three groups. Conclusion: this experiment confirmed that the acupuncture at Lian Quan point can not only induce the changes in the discharges of the swallowing muscle directly, but also increase the expression of c-fos protein in the nucleus of the solitary tract, and further indicate that the nucleus of the solitary tract is involved in the regulation of the swallowing movement and plays an important role in the acupuncture.
【學(xué)位授予單位】:廣州中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】:博士
【學(xué)位授予年份】:2015
【分類號(hào)】:R245-0
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