针刺任督脉经穴对脑干吞咽中间神经元的调节作用

发布时间：2018-05-13 21:45

本文选题：针刺 + 吞咽　；参考：《广州中医药大学》2015年博士论文

【摘要】：吞咽是人类最为基本和重要的生理功能之一,健康人每天进行无意识的吞咽活动可达一千次,其主要作用是将食物由口咽部传递至食管。其整个过程需要6对颅神经和25块肌肉协同完成。吞咽是一个复杂的感觉运动活动,依靠大脑皮层、脑干吞咽中枢和颅神经(Ⅴ、Ⅸ、Ⅹ、(?) (?))之间高度系统性的相互作用。这一过程既有主动成分又有非主动成分,分别反应了皮层和脑干的中枢调节途径。针刺作为一种在临床上治疗吞咽障碍、调节吞咽功能行之有效的方法,其内在机理一直未有明确的认识,近20年来,围绕吞咽反射的脑干神经元网络所开展的大量研究使得我们对吞咽的生理学机制有了一个较为清晰的认识,脑干吞咽神经元网络主要由传入纤维、中间神经元和运动神经元组成。针刺作用在吞咽脑干神经元网络的机制尚未清楚,这些研究的基础为我们研究针刺调节吞咽功能提供了一定的思路,因此,本研究针对脑干背侧吞咽中枢,探讨针刺对该吞咽初级中枢的作用机制。目的：观察针刺对脑干背侧吞咽中间神经元的激活情况,从生理状况下神经电生理情况、损毁核团后针刺效应变化以及针刺对神经解剖形态学的效应几个方面,来探讨针刺对初级中枢吞咽中间神经元的作用,揭示针刺调节吞咽功能的可能机理。实验一针刺对吞咽相关中间神经元(孤束核)放电的调节方法：SD大鼠55只,雌雄不限,水合氯醛麻醉后,先进行前颈部手术,暴露下颌舌骨肌以及喉上神经,分别连接记录电极以及刺激电极,再进行脑部手术,颅骨钻孔后暴露延髓闩部,用立体定位仪将玻璃微电极插入延髓孤束核内,开始记录神经元放电,先刺激喉上神经引发肌电变化及吞咽,确定吞咽相关神经元后,分别针刺风府穴、廉泉穴、内关穴以及足三里穴30s,观察神经元放电变化以及肌电变化。记录完毕后标记记录位置,进行组织学鉴别,定位不准确的数据弃之不用。结果：本实验共记录到时相型吞咽相关神经元共63个,对针刺“风府”穴起反应的神经元共41个(65%),“廉泉”穴36个(57%),“内关”穴23个(37%),“足三里”穴19个(30%)。自发放电型神经元共完整记录到107个,与吞咽运动相关的神经元共记录到70个,主要为表现为兴奋性神经元。针刺“风府”穴后,NTS神经元放电频率增加的共49个,减少的共6个,无变化的共15个；针刺“廉泉”后NTS神经元放电频率增加的共53个,减少的共5个,无变化的共12个,针刺“内关”后NTS神经元放电频率增加的共38个,减少的共3个,无变化的共29个,针刺足三里后,NTS神经元放电频率增加的共31个,减少的共11个,无变化的共28个。针刺以上四穴对孤束核吞咽相关神经元的激活作用,以廉泉穴及风府穴较为显著,内关与足三里对吞咽相关孤束核神经元均有-定程度的激活作用,其中对各组兴奋性神经元放电频率变化率进行比较,针刺廉泉穴及风府穴与针刺其他穴位分别比较,差异具有统计学意义(P0.05)。结论：针刺后可诱发吞咽相关神经元的放电变化,表明针刺对脑干背侧吞咽中枢神经元具有激活作用,在穴位的选择上,廉泉穴与风府穴对神经元激活作用明显,内关穴及足三里穴也可一定程度的激活该神经元。共12个,针刺“内关”后NTS神经元放电频率增加的共38个,减少的共3个,无变化的共29个,针刺足三里后,NTS神经元放电频率增加的共31个,减少的共11个,无变化的共28个。针刺以上四穴对孤束核吞咽相关神经元的激活作用,以廉泉穴及风府穴较为显著,内关与足三里对吞咽相关孤束核神经元均有-定程度的激活作用,其中对各组兴奋性神经元放电频率变化率进行比较,针刺廉泉穴及风府穴与针刺其他穴位分别比较,差异具有统计学意义(P0.05)。实验二损毁孤束核对针刺任督脉经穴调节吞咽肌运动的影响方法：SD大鼠42只,分为风府组、廉泉组以及内关组,每组均分为损毁和假损毁两部分,麻醉后进行颈部手术同实验一,仅放置肌电记录电极,脑部手术与实验一相同,先对各组进行针刺60s,观察下颌舌骨肌肌电放电及吞咽频数,然后在立体定位仪引导下将金属电极插入孤束核相关核团,通阳极直流电0.1mA,时间20s,进行核团电损毁。损毁结束后再次针刺以上三组穴位,观察肌电变化以及吞咽频数。假损毁实验中,仅将电极插入核团,不进行通电,时间也为20s。实验结束后,对延髓进行组织学检测,损毁部位不准确者数据弃之不用。结果：在损毁孤束核前,针刺“廉泉穴”组最多引发的下颌舌骨肌放电频数,较风府组与内关组均有显著差异(P0.05),而针刺“风府”穴与“内关”穴两组比较无明显差异(P0.05)。损毁前后进行针刺,以上三组进行损毁前后组内比较,损毁后针刺引发的下颌舌骨肌肌电放电频次较损毁前明显减少,其中以廉泉穴组以及风府穴组变化最为明显,损毁前后具有明显差异(P0.01),针刺内关穴在损毁前后吞咽频次差异也具有统计学意义(P0.05)。损毁孤束核后对三组进行组间比较,针刺不同穴位对吞咽的影响比较无明显差别(P0.05)。假损毁对照试验显示,针刺风府穴、廉泉穴以及内关穴,在假损毁前后诱发的吞咽频数无明显差异结论：电解损毁孤束核后,针刺“廉泉”、“风府”以及“内关”穴对大鼠的吞咽诱发作用明显减弱,提示孤束核是针刺调节吞咽功能中的一个重要的初级中枢。针刺廉泉穴对吞咽的诱发作用明显,风府穴及内关穴也可在一定程度上诱发吞咽运动,提示穴位的选择对针刺调节吞咽功能具有一定的意义。实验三针刺对孤束核中c-fos蛋白表达的影响方法：SD大鼠24只,分为空白组、风府组、廉泉组以及内关组,常规麻醉后,进行颈部手术,暴露下颌舌骨肌后插入记录电极,分别电针上述三组穴位,频率10Hz,2mA,共20min,同时记录下颌舌骨肌肌电变化以及吞咽运动,针刺完毕后,对大鼠进行灌流取脑,保留延髓及部分脊髓,固定脱水后进行冰冻切片,并用免疫组化方法观察大鼠延髓相应区域的c-fos蛋白表达变化。结果：电针各组大鼠诱发吞咽肌放电以针刺廉泉穴及风府穴较为显著,效果由于内关组(P0.05),空白组则无吞咽运动产生。针刺后大鼠各组c-fos蛋白阳性表达方面,在孤束核区域范围内,空白组在孤束核区域内可见零星的c-fos阳性表达细胞,其余三组均可见阳性表达的细胞,其中以廉泉穴组、风府组以及内关组均有较多的阳性表达细胞表达,三组比较无显著差异。结论：本实验证实了针刺廉泉穴、风府穴不仅可以直接诱发吞咽肌放电变化,还可使孤束核内c-fos蛋白表达升高,从形态学的角度进一步表明了孤束核参与了针刺调节吞咽运动并起到重要的作用。
[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.

【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】：2015
【分类号】：R245-0

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