| Objective:This research started from the Pirt-TRPV1 pathway to observe the effects of acupuncture on the expression of Pirt in dorsal root ganglion(DRG)and TRPV1 in colonic mucosa of rats with IBS-D,and to analyze the relationship between acupuncture relief of visceral hypersensitivity(VHS)and the Pirt-TRPV1 pathway,to explore the mechanism of acupuncture alleviating VHS,serving as a scientific evidence for acupuncture in IBS-D treatment.Methods:40 healthy clean grade adult Sprague Dawley(SD)rats,with half males and half females,were conducted to a pre-modeling behavioral evaluation after 7-day adaptive feeding to remove the incompetent rat.Then qualified rat were stratified by body weight and randomly assigned into control group,model group,model + acupuncture group(acupuncture group),and model + pinaverium bromide group(medicine group)by lottery.Then the IBS-D rat model in the model group,acupuncture group,and medicine group was established through the combination of chronic unpredictable mild stress(CUMS)and the administration by gavage of senna soaking agent for 14 days.Then a model test was conducted to remove the incompetent rat again.Then the rats which were determined to be successful under the model further entered the 14 days’ treatment stage.Rats in the acupuncture group were alternatively electro-acupunctured at ST25(Tianshu),ST36(Zusanli),SP6(Sanyinjiao)and LR3(Taichong)on both sides,lasting 15 minutes for once a day.The medicine group was treated by the intragastric method with a volume of 10 mL/kg of the 2.7mg/mL pinaverium bromide.A behavioral evaluation was conducted after treatments.The changes of rats’ VHS were estimated by the pain threshold resulted from the abdominal withdrawal reflex(AWR)test,changes of diarrhea were estimated by diarrhea index,and anxiety and depression-like behaviors were estimated by OT% in the elevated plus maze test and sucrose preference(SP%)respectively.Following the above-referenced evaluation,immunohistochemistry(IHC)was used to evaluate the expression of Pirt in DRG and colonic mucosa TRPV1 of rats in all four groups.Results:1.General stateBefore modeling,there was no significant difference in food intake and body weight between the four groups(P>0.05,P>0.05).After and before modeling,there was no significant difference in food intake of control group(P>0.05),and its weight gained apparently(P<0.01).The food intake in the model group,the acupuncture group and the drug group was significantly reduced(P<0.01)and weight gained slowly(P>0.05).After intervention and after modeling,there was no significant difference in the food intake of control group(P>0.05),and weight gained significantly(P<0.01).There was no significant difference in the model group’s food intake(P>0.05),and the body weight was apparently reduced(P<0.01).The food intake of acupuncture group and medicine group increased apparently(P<0.01),and weight increased significantly(P<0.01).After modeling,the food intake of rats in the model group,the acupuncture group and the medicine group decreased significantly compared with the control group(P<0.01),and weight gained slowly(P>0.05).After treatment,the food intake and body weight gain of the acupuncture group and the drug group were significantly higher than the model group(P<0.01,P<0.01).2.Visceral sensitivityBefore modeling,there was no significant difference in the visceral pain threshold of rats in four groups(P>0.05).There was no significant difference in visceral pain threshold of control group(P>0.05).After and before modeling,the visceral pain threshold of the model group,the acupuncture group and the medicine group was obviously reduced(P<0.01).After intervention and after modeling,there was no significant difference in visceral pain threshold of model group(P>0.05),while there was a obvious improvement in the acupuncture group and the medicine group(P<0.01).After modeling,the visceral pain threshold of the model group,the acupuncture group and the medicine group was obviously reduced(P<0.01)than it in the control group,meaning an obvious increase in visceral sensitivity.After treatment,there was a obvious improvement in the visceral pain threshold of the acupuncture group and the medicine group(P<0.01)than in the model group,meaning an obvious decrease of visceral sensitivity.3.Diarrhea indexBefore modeling,the rats in the four groups had no diarrhea,and all the diarrhea index was 0.The control group didn’t have diarrhea during the whole period and the diarrhea index was 0.After and before modeling,the diarrhea index of the model group,the acupuncture group and the medicine group was obviously increased(P<0.01).After intervention and after modeling,there was no significant difference in diarrhea index of model group(P>0.05),the acupuncture group and the medicine group was obviously reduced(P<0.01).After modeling,the diarrhea index of the model group,the acupuncture group and the medicine group was significantly increased(P<0.01)compared with the control group.After treatment,the diarrhea index of the acupuncture group and the medicine group was significantly decreased(P<0.01)compared with the model group.4.Anxiety and depression-like behaviorsBefore modeling,there was no significant difference in OT% and SP% of four groups rats(P>0.05,P>0.05).There was no significant difference in OT% and SP% of control group(P>0.05).After and before modeling,the OT% and SP% of the model group,the acupuncture group and the medicine group was obviously reduced(P<0.01,P<0.01).After intervention and after modeling,there was no significant difference in OT% and SP% of model group(P>0.05,P>0.05),while there was a obvious improvement in the acupuncture group and the medicine group(P<0.01,P<0.01).After modeling,the OT% and SP% of the model group,the acupuncture group and the medicine group were significantly decreased(P<0.01,P<0.01)compared with the control group.After treatment,there was a manifest difference in the OT% and SP% of the acupuncture group and the medicine group(P<0.01,P<0.01),than it in the model group.5.Pirt and TRPV1The mean optical density of Pirt and TRPV1 of rats in the model group is remarkably increased(P<0.01,P<0.01)compared with the control group;while the mean optical density of Pirt and TRPV1 of the acupuncture group and the medicine group is noticeably decreased(P<0.01,P<0.01)compared with the model group.Conclusions:1.Acupuncture at ST25,ST36,SP6 and LR3 can effectively reduce the VHS of IBSD rats,releasing their diarrhea and improving anxiety and depression-like behaviors,proved as one of the effective methods for IBS-D treatment.2.The forming mechanism of IBS-D VHS might be related to the access between Pirt and TRPV1,which is possibly restrained by acupuncture,leading to the release of IBS-D VHS and acupuncture’s function in IBS-D treatment. |
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