| ObjectiveIn this experiment,irritable bowel syndrome(IBS)rats are the researched object,by means of multi channel synchronous recording technology,fluorescent PCR and immunohistochemistry,to observe the effect of Electroacupuncture "PC6" and "BL25"acupoint on IBS rats intestinal sensitivity,emotional psychological activity and the expression of TRPV1 and ICC in the gut effect of combining the ventroposterolateral nucleus of rats IBS model(VPL)and the change of discharge of neurons of the GalR1 region,to investigate the effect of electroacupuncture at different acupoints on the difference of IBS and central analgesia mechanism.MethodsAfter healthy pregnant Wistar rats birth rats,they were randomly divided into blank control group and model group,except the blank control group,the maternal separation and acetic acid enema combined with colorectal balloon stimulation combined preparation of intestinal irritable bowel syndrome rat model.After modeling,the model group was randomly divided into model control group,electro acupuncture "PC6" group and electro acupuncture "BL25" group,15 in each group.Stay in length in rats at 8 weeks of age were randomly selected from each group to electrode implantation surgery rats in each group to ensure success of 3,recovery of the rats after 1 weeks,the control group and model control group rats did not do acupuncture treatment,acupuncture treatment of electroacupuncture"PC6" group and electroacupuncture group were "BL25" the corresponding points,once every other day,a total of 5 times.After the end of treatment,according to the classification of Bristol were observed and recorded the rats stool rank score;the intestinal sensitivity of rats was evaluated by Abdominal withdrawal reflex experiment,observation,and the number of initial wave latency of contraction wave pressure;change by open field test emotional behavior,the number of open field cabinet level observation and the vertical movement of the body;used in multi channel synchronous recording technique to observe the brain VPL neurons change;by fluorescence quantitative PCR(q-PCR)method to detect 1 galanin receptors within the thalamus(GalR1)positive expression;immunohistochemical method was used to detect the colon capsaicin receptor(TRPV1)and interstitial cells of Cajal(ICC)c-kit receptor expression.Results1 Effects of electroacupuncture on the stool and behavioral responses in IBS ratsCompared with the blank control group,model control group rat fecal Bristol score increased significantly(P<0.01);Abdominal withdrawal reflex experiment showed that pain response in rats significantly shorten the incubation period,a contraction wave number was significantly increased,the initial value of wave pressure decreased significantly(P<0.01,P<0.01,P<0.01);open field box the horizontal and vertical movements were significantly reduced(P<0.01,P<0.01);Electroacupuncture "Dachangshu" group of vertical movement was significantly reduced(P<0.05).Compared with the model control group,electroacupuncture at "PC-6" and "BL-25"rats stool Bristol scores were significantly decreased(P<0.01,P<0.01);Electroacupuncture group rats pain prolonged latency,reducing the number of contraction wave and initial wave pressure increased significantly(P<0.05,P<0.05,P<0.01).EA "BL25" group rats pain latency prolonged significantly,significantly reduced systolic wave number and initial wave pressure increased significantly(P<0.01,P<0.01,P<0.01);Electroacupuncture at "PC6"and "BL-25" group level movements increased significantly(P<0.01,P<0.05).2 Effects of electroacupuncture on VPL neuron discharge in IBS ratsCompared with the blank control group,the discharge frequency of the neurons in the model group was significantly increased(P<0.01),and the discharge frequency of the neuron in the EA "BL-25" group was significantly increased(P<0.05);Compared with the model control group,EA "PC-6" the discharge frequency of type I neurons in VPL group was significantly reduced(P<0.01),and EA "BL25" the discharge frequency of VPL was significantly reduced(P<0.05).3 Effects of Electroacupuncture on the expression of GalR1,TRPV1 and c-kit in the thalamus of IBS model ratsCompared with the blank control group,model control group GalR1 showed no significant change(P>0.05),and TRPV1 and c-kit expression levels were significantly increased(P<0.01,P<0.01),electroacupuncture "BL25" group c-kit expression levels were significantly increased(P<0.01).Compared with the model control group,there was no significant change of GalR1 in the electroacupuncture group(P>0.05),while the positive expression of TRPV1 and c-kit was significantly lower(P<0.05,P<0.01).Compared with electroacupuncture EA-PC6 group,the positive expression of TRPV1 and c-kit in EA-BL25group was significantly lower(P<0.01,P<0.01).Conclusion(1)preparation of the model rats,the change of character of stool,intestinal sensitivity increased and psychological abnormalities,with the onset of IBS features,suggesting that maternal separation with acetic acid enema combined with intestinal dilatation can be successfully replicated IBS model.(2)EA can improve the rat's stool character,emotional psychology and relieve abdominal pain.Electroacupuncture of "Dachangshu" point to reduce IBS rat intestinal sensitivity,relieve pain symptoms better than EA Neiguan;Electroacupuncture at Neiguan point on the abnormal psychological symptoms improve mood better than EA"Dachangshu" point.It is suggested that electroacupuncture at "Neiguan" and "Da Yu Shu"points to improve the symptoms of IBS,which reflects the specificity of acupoints.(3)the excitability of rat model of IBS VPL ? and ? class two neurons increased,the discharge frequency increased after electroacupuncture;found the excitability of VPL ? and? two neurons decreased,the discharge frequency is reduced;the type ? neurons decreased frequency of electroacupuncture after a greater extent than the large intestine Shu,and Electroacupuncture Dachangshu points lower frequency of class ? neurons are superior to that of neiguan.It is suggested that the regulation of the VPL in the Neiguan and the large intestine Yu Erxue may be different from another angle.(4)the contents of TRPV1 and ICC in the colon of IBS model rats were increased abnormally,and the contents of TRPV1 and c-kit were decreased after electroacupuncture,and the content of TRPV1 and ICC was significantly better than that of Neiguan point.It is suggested that the mechanism of electroacupuncture relieving abdominal pain in IBS rats may be related to the regulation of intestinal TRPV1 and ICC,and there are differences in the regulation of two points. |
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