| Objective:The purpose of the study is to investigate the physiological condition of electroacupuncture at "Lian Quan" and "Fengfu" on the swallowing function and the influence of brainstem motor neurons in the nucleus ambiguus swallowing functionof SD rats and to study the possible role of regulating neurotransmitter brainstem swallowing neurons in the network in electroacupuncture that reveal the acupuncture physiological mechanism of adjusting swallowing functionfrom the aspects of electrophysiological and material foundation.Method:Experiment 1:Regulation of the Electroacupuncture at "Lian Quan" and"Feng Fu" in the deglutition and its motor neuronsThe experiments were performed on 32 healthy male or female Sprague-Dawley(SD)rats.The rats were anesthetized(intraperitoneal)with 20%urethane(0.5ml/100g).The neck surgery was operated to separate the recurrent laryngeal nerve which connected the stimulating electrode and to exposed the mylohyoid muscle which connected the recording electromyogrphy electrodes and.Then the rats were fixed in the stereotaxic apparatus and on the brain surgery.The medulla was exposed bolt and the tungsten electrode was inserted into the medullary nucleus ambiguus with the electric propulsion.And the discharges of neurons was recorded.The laryngeal nerve was stimulated to determine whether neurons recorded for swallowing motor neurons.In order to determine whether the swallowing related neurons,double distilled water was injected to the throat inducing swallowing.The acupoint of Lian Quan,Fengfu,Neiguan and Zusanli were applied on electroacupuncture for lmin to observe the changes of neuronal discharge after acupuncture.And the rat were in the same method to induceswallowing again.The number of swallows and the latency from the onset of infusion to the first swallow were observedafter the electroacupuncture and.After the record was completed,the location of the recording was damaged by the electricity,and the histological identification was carried out.If the location was inaccurate,the data were discarded.Experiment two:the effect of chemical lesion of NA on the swallowing activities after EA at fengfu and lianquan80 SD rats,male and female,were randomly divided into IBO chemical damage group and pseudonuclear damage group.After anaesthetized,the rats of chemical damage group ware microinjected in bilateral nucleus with 0.2 ?L anserine acid(0.4 g/mg)and pseudonuclear damage group was microinjected with 9%physiological saline.We compared the effect of the electroacupuncture at Lian Quan point,Feng Fu point,Neiguan point and Zusanli point on the swallowing function before and after injection.After the record was completed,the location of the recording was located and the histological identification was carried out.If the location was inaccurate,the data were discarded.Experiment three:the effect of microinjection of different drugs on the swallowing activity of electroacupuncture on "Lian Quan" and "Feng Fu" in rats96 SD rats,male and female,were randomly divided into four groups,including the Lian Quan Group,the Feng Fu Group,the Neiguan group and the Zusanli group,with 24 rats in each group.After anaesthetized,DDW would be automatically infused into the pharyngolaryngeal region.0.2 ?LCNQX(10mM),APV(50mM),Muscimol(10mM),physiological saline were injected in bilateral nucleus ambiguus,respectively.the electroacupuncture was carried out lmin after the injection.When the electroacupuncture was end,DDW would be automatically infused into the pharyngolaryngeal region again.After the record was completed,the location of the recording was located and the histological identification was carried out.If the location was inaccurate,the data were discarded.Experiment four:the effect of Electroacupuncture of "Lian Quan" and "Feng Fu" on the expression of c-Fos protein in the nucleus ambiguus50 SD rats were randomly divided into four groups,including the Lian Quan Group,the Feng Fu Group,the Neiguan group,the Zusanli group and the blank group,with 10 rats in each group.The parameter of electroacupuncture was in the continuous wave,current intensity 2mA,frequency 2Hz,wave width 2ms,duration 20min,and the reference electrode was distance about 1mm about point.The blank group was not given the intervention of electroacupuncture.After electroacupuncture at corresponding points of 20min in each group,the rats were perfused and removed for preservation of the medulla oblongata.After frozen dehydration,the expression of c-Fos protein in the suspected nucleus was observed by immunofluorescence.Results:1.Electroacupuncture at "Lian Quan" and "Feng Fu" point can promote swallowing activity,increasing the frequency of the hyoid muscle discharge and shorten the first swallow latency(P<0.01),while the electroacupuncture of "Neiguan" and "Zusanli" have no obvious effect.The electroacupuncture of"Lian Quan" and "Feng Fu" were the main activation effect(P<0.01)on the deglutition motor nucleus ambiguus,with the increasing frequency of discharges of neurons in the nucleus ambiguus.2.After the chemical destruction,the electroacupuncture of "Lian Quan"and "Feng Fu" acupoint had obviously weakened the swallowing function of rats.In the blank group without electroacupuncture intervention,the activity of swallowing was significantly reduced(P<0.01)compared with the false lesion after IBO chemical damage.In Lianquan electroacupuncture intervention group,IBO in the chemical lesion of bilateral nucleus ambiguus after electroacupuncture at "Lian Quan" and "Fengfu","Neiguan" and "Zusanli",significantly reduce the damage before swallowing activity than before EA(P<0.01).While false damage did not affect the regulatory effect of Electroacupuncture on swallowing(P>0.05).3.After microinjection of glutamate receptor antagonist CNQX,APV,GABAa receptor agonist muscimol on the bilateral nucleus ambiguus respectively,they inhibited the regulation of the swallowing activities of the "Lian Quan" and"Feng Fu" points in the electroacupuncture.The microinjection of APV and CNQX is more inhibited than that of muscimol.4.After electroacupuncture of "Lian Quan" or "Feng Fu" point,more c-Fos protein expression products(P<0.01)were found in the rat's motor neurons.Conclusion:1.There is a relative specific connection between the "Lian Quan","Feng Fu" and the swallowing function2.Nucleus ambiguous is the important primary center of "Lian Quan","Feng Fu" and swallowing connection.It participates in regulating the swallowing function of electroacupuncture,and is related to the activity of swallowing muscles under transmission related information.3.Glutamate and GABA are involved in the regulation of the deglutition activity through the Electroacupuncture of the receptor,and the glutamate receptor is more obvious.4.Acupuncture at Lianquan or Fengfu can be transmitted to the motor neuron impulse swallowing nucleus ambiguus,activating the neurons and expressing the c-Fos immediate specific reaction protein.It shows that the nucleus is an important center for the regulation of electroacupuncture.provides the neural morphological evidence,providing the evidence of neural morphology. |
PDF Full Text Request