| ObjectiveThis study tried to choose IBS rats as the research medium,and observe the effects of electroacupunture(EA)at "Zusanli"(ST 36),"Hegu"(LI 4)and "dachangshu" point(BL 25)on intestinal sensitivity,motility and the expression of the change of 5-HT3AR in colon tissues.And to combine the change of characteristic of neuron discharge in the region of VPL and DMV of the brain.To discuss the treatment effects and differences of points on different Meridians and different nerve segment to treat the same disease.To illustrates the exist of Specificity of Therapeutic Effects of Acupoints,and analyze the relevant mechanism.Methods80 neonatal wistar rats were randomly divided into the blank control group,the model control group,the "ST 36"group,the "LI 4" group and "BL 25" group,16 in each group.In addition to the blank control group,the other groups were prepared by neonatal maternal separation and acetic acid enema combined with colon and colorectal distension(CRD)stimulation.After 2 months,rats were randomly selected from each group to embed electrodes in VPL and DMV,3 rats per group,per brain nucleus.One week after the operation,"ST 36"group,"LI4" group and the "BL 25" group were given electroacupuncture(EA)intervention once every other day for 5 times.The blank control group and the model control group were not intervened.After operation,Bristol Stool Scale was used to observe the faeces types of the IBS rats before and after electroacupuncture;Abdominal withdrawal reflex(AWR)was recorded to evaluate the intestinal motility and sensitivity;The expression of 5-HT3AR in colon(Mucosal and muscular layer)was detected by immunohistochemistry;To observe the changes of VPL and DMV neurons by multichannel in vivo recording technique.Results1.The effects of electroacupuncture at different acupoints on stool traits of IBS model rats.Before electroacupuncture:compared with the blank control group,the stool property score of the model control groups was significantly increased(P<0.01);After electroacupuncture:compared with the model control group,the stool score of there treatment groups was was significantly decreased(P<0.01).Compared with the "ST 36" group,the Bristol stool score of the "LI 4" group increased(P<0.05);compared with the "LI 4" group,the Bristol stool score of the "BL 25" groups decreased(P<0.05),and there was no significant differencebetween these two groups.2.The effects of electroacupuncture at different acupoints on abdominal withdrawal reflex(AWR)of IBS model rats.Compared with the blank control group,the latency was significantly shortened(P<0.01)and the number of contraction waves was significantly increased(P<0.01)in the model control:group.Compared with the model control group,the latency was obviously prolonged(P<0.01)and the number of contraction waves was decreased(P<0.01,P<0.05)in the three treatment groups.Compared with the "ST 36" group,the latency was prolonged in "LI 4" group and "BL 25" group(P<0.05).3.The effects of electroacupuncture at Different acupoints on the immunohistochemical expression of 5-HT3A receptor in colon’s mucosal layer of IBS model rats.Compared with the blank control group,the immunohistochemical expression of 5-HT3A receptor in mucosal layer of colon was significantly increased in the model control group(P<0.01),which explained that the IBS model was successful.Compared with the model control group,the immunohistochemical expression of 5-HT3A receptor in mucosal layer of colon was decreased in the there treatment groups.Compared with "ST 36" group,the expression of 5-HT3A receptor was obviously decreased in "LI 4" group,while the expression of 5-HT3A receptor was obviously increased in "BL 25" group compared with "LI 4" group.4.The effects of electroacupuncture at Different acupoints on the immunohistochemical expression of 5-HT3A receptor in colon’s mucosal layer of IBS model rats.Compared with the blank control group,the immunohistochemical expression of 5-HT3A receptor in muscularis of colon was significantly increased in the model control(P<0.01),which explained that the IBS model was successful.Compared with the model control group,the immunohistochemical expression of 5-HT3A receptor in muscularis of colon was decreased in the three treatment groups(P<0.01).Compared with "ST 36" group,the immunohistochemical expression of 5-HT3A receptor in muscularis of colon was significantly decreased in "LI 4"group and "BL 25" group(P<0.01).5.Effect of electroacupuncture at "ST 36","LI 4" and "BL 25" on the discharge of VPL neurons in IBS model ratsUnit Ⅰ:Compared with the blank control group,In the model control group,the discharge frequency of both unit I neurons in VPL were increased(P<0.05).Compared with the model control group,the VPL discharge frequency of the ST 36 group and LI 4 group were reduced(P<0.05).Unit Ⅱ:Compared with the blank control group,the discharge frequency of unit Ⅱneurons in the model control group was increased significantly(P<0.01).Compared with the model control group,the firing frequency of neurons in LI 4 and BL 25 groups were reduced(P<0.01,P<0.05).Compared with ST 36 group,the firing frequency of neurons in both LI 4 and BL 25 groups were increased(P<0.05).6.Effect of electroacupuncture at "ST 36","LI 4" and "BL 25" on the discharge of DMV neurons in IBS model ratsUnit Ⅰ:Compared with the blank control group,the discharge frequency of unit Ⅰ neuron was increased significantly in the model group(P<0.01),and increased in both LI 4 group and BL 25 group(P<0.05).Compared with the model control group,the discharge frequency of unit Ⅰ neurons was decreased in ST 36 group(P<0.05).Unit Ⅱ:Compared with the blank control group,the discharge frequency of unit Ⅱ neurons in the model control group was increased significantly(P<0.01),and the discharge frequency of unit Ⅱ neurons was increased in the BL 25 group(P<0.05)Compared with the model control group,the discharge frequency of unit Ⅱ neurons in ST 36 group was decreased(P<0.05).Conclusion1.In this experiment,the IBS model rats developed abdominal pain and diarrhea by co-preparing the model.The stool property of IBS model rats become dilute and moist,intestinal sensitivity was increased and the motility was disordered,which showed that the combined model of maternal and infant separation and acetic acid ascending enema combined with colon dilatation and stimulation can successfully replicated a stable IBS-D rat model in this study.2.Electroacupuncture at "ST 36","LI 4" and "BL 25" can improve fecal traits,inhibit high intestinal sensitivity and dynamic abnormalities in IBS model rats,In other words,alleviate symptoms such as abdominal pain and diarrhea.Among them,"LI 4" has the strongest regulating effect on intestinal sensitivity."ST 36" and "BL 25" have obvious effects on improving stool traits and regulating intestinal motility.Which can suggest that different acupoints can treat different conditions of the same disease,but their regulatory effects are different.And it is further confirmed that the acupoint effect is specifically present and has relativity and integrity.3-The expressions of 5-HT3A receptors mucosal and muscular layers of the colon in IBS model rats were significantly enhanced,and electroacupuncture at the three acupoints could reduce the expression of both.In addition,"LI 4" regulated mucosal 5-HT3A receptors better than "ST 36" and "BL 25",and "ST 36" regulated muscular 5-HT3A receptors better than other two acupoints.It was suggested that 5-HT3A receptors may be involved in mediating the occurrence of intestinal hypersensitivity and dyskinesia in IBS rats,and electroacupuncture can relieve abdominal pain and diarrhea by down-regulating the positive expression of its receptor.4.The neuronal firing patterns of VPL and DMV in IBS model rats had changed.Electroacupuncture at all the three acupoints can inhibit the discharge of unit Ⅰ neurons in VPL and unit Ⅰ and Ⅱ neurons in DMV of IBS model rats,and excite the discharge of unit Ⅱ neurons in VPL,Among them,"ST 36" has the strongest excited effect on the discharge frequency of unit Ⅱ neurons in VPL and inhibitory effect on the discharge frequency of unit Ⅰ and Ⅱ neurons in DMV,and "ST 36" has better inhibitory effect on the discharge than "LI 4" and "BL 25",which further confirmed the central level of meridian effect specificity. |
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