The Effect Of Electroacupuncture At Different Points On The Discharge Of ACC Neurons And The Expression Of NMDA Receptors In IBS Rats

Chinese medicine have the view that man is the unity of "physical and spirit",various life activities are led by "spirit" and emotion is the external manifestation of "spirit" on the basis of Zang Fu organs,meridians and collaterals.Because of the role of emotion in the occurrence and development of the disease can not be ignored,traditional Chinese medicine special emphasis on "spirit".Acupuncture,as a special therapy of Chinese medicine,attach importance to both " physical and spirit" to a whole effect of man,which can adjust the Qi of meridians and collaterals by stimulating the acupuncture points to keep the balance of the body.Irritable bowel syndrome is a typical psychosomatic disease,Zusanli and Neiguan point are usually used in clinical treatment and has a good effect on it."Tianshu" is local acupoints and good for gastrointestinal diseases,while,"Neiguan"and "Zusanli" are distal acupoints,they are have the effect on both " physical and spirit,dieases but the emphases is different.From the perspective of modern medicine,the three points belong to different nerve segments;then,whether there are differenees between the effects of these three points for IBS in the pain-emotion?objectiveIBS rats are the media of study in this experiment,observing the change of ACC neurons'spike and the expression of NMDA receptors by electro acupuncture on different acupoints to explore the effect of different acupoints impact on IBS rats,compare the difference and try to find the possible central mechanism.MethodWistar rats were randomly divided into two groups:blank control group and model preparation group.The model preparation group was prepared by the method of "separation of mother and child and acetic acid enema" and "colorectal distension stimulation".After the modeling success,the model preparation group were randomly divided into model group,Neiguan group,Tianshu group and Zusanli group.The rats in the treatment group were treated with electroacupuncture at 9 weeks age,once every other day for a total of 5 times.After the end of treatment,The visceral sensitivity of rats was evaluated by AWR,observed latency and number of contraction waves;the emotional and psychological behavior of rats were evaluated by OFT,and observed the horizontal and vertical movements;The changes of ACC's neuron pike in brain were observed by in vivo multi-channel synchronous recording technique;and the expression of NMDA receptor in colon and ACC was detected by immunohistochemistry and fluorescence quantitative PCR.Result1.the effect of electroacupuncture at different acupoints on the behavior of IBS model rats 1)Effect of electroacupuncture at different acupoints on IBS model rats in AbdominalwithdrawalCompared with the blank group,the latent period of the model group was significantly shortened and the number of contraction wave was significantly increased(P<0.01)_Compared with the model group,the latency were significantly prolonged(P<0.01)in the treatment group,and the number of systolic wave was significantly reduced inTianshu and Zusanli group(P<0.01).Compared with the Tianshu group,systolic waves in Zusanli group were more(P<0.05),and Neiguan group' waves were most(P<0.01),and the latency in Neiguan group was shorter than Tianshu(P<0.05).2)Effect of electroacupuncture at different acupoints on open field test of IBS model ratsCompared with the blank group,the rats in the model group,the number of horizontal movement and vertical movement were significantly reduced(P<0.01).Compared with the model group,the number of horizontal movement was significantly increased in Neiguan group(P<0.01),the number of vertical movement was increased(P<0.05);Tianshu group and Zusanli group were only increased in the number of vertical movement(P<0.05).2.the effect of electroacupuncture at different acupoints on ACC neurons in IBS ratsCompared with the blank group,the discharge frequency of Inhibitory neurons in model group was decreased(P<0.05),and the discharge frequency of Excitatory neurons was significantly increased(P<0.01).Compared with the model group,the discharge frequency of Inhibitory neurons in Neiguan group and Zusanli group was increased(P<0.05),and the discharge frequency of Excitatory neurons in three groups were significantly reduced(P<0.01).3.Effects of electroacupuncture at different acupoints on NMDAR in IBS rats1)Effects of electroacupuncture at different acupoints on the expression of NMDAR in ACC of IBS ratsCompared with the blank group,the NR1 and NRZA in ACC in the model group were significantly increased(P<0.01),and the NR2B was increased(P<0.05).Compared with the model group,NR2A,NR2B NRI were decreased in Neiguan group(P<0.01,P<0.05)NR1,NR2B were reduced in Zusanli group(P<0.05);Tianshu group only NR1 decreased(P<0.05).2)the effect of electroacupuncture at different acupoints on the expression of NMDAR immunoreactive substance in colon of IBS ratsCompared with the blank group,the expression of NR1 and NR2B positive immunoreactivity in the colon of model group was significantly increased(P<0.01).Compared with the model group,the expression of NR1 positive immunoreactivity was decreased in three groups(P<0.05),while the Zusanli group was significantly decreased(P<0.01);the expression of NR2B positive immunoreactivity in Tianshu group and Zusanli group were reduced(P<0.05),Neiguan group although reduced,but no significant difference between two groups.conclusion1.IBS model rats with abdominal sensitivity pain and emotional and psychological abnormal behavior,consistent with the pathological features of IBS,which proved that,mother separation plus acetic acid enema combined with colorectal balloon" stimulation was successfully prepared IBS rat model.2.Electro acupuncture "Neiguan" Tianshu" and "Zusanli" point can reduce the intestinal sensitivity,relieve physical pain symptoms,improve abnormal mental emotional behavior of IBS model rats.But the effect of alleviate abdominal pain of IBS,Tianshu was best,Zusanli was better than Neiguan Acupoint;the effect of improve mood,Neiguan was best,Zusanli was better thanTianshu point.The difference of the three point effect may be related to the difference of the acupoints and the distribution of the nerve segment.3.The frequency of Inhibitory neurons were decreased and The frequency of Excitatory neurons were increased in IBS rats.After electroacupuncture,two unit were regular.the effect of Neiguan is best and Zusanli is better.The results showed that the abnormal activity of ACC neurons in the brain region was related to the IBS model,and the central mechanism of electroacupuncture relieving pain was probably related to the regulation of ACC neurons.4.The expression of NMDA receptor was incrased both in ACC and colon,while the expression was decreased after acupuncture,and the Neiguan's effect was best and Zusanli was better in ACC;while Zusanli was best and Tianshu was better in colon.The results showed that NMDA receptor may be an important factor mediating visceral pain and pain in IBS,and electroacupuncture could relieve the pain and aversive emotion of IBS by decreasing the content of NMDA receptor.