Effect Of The Suppressing Mu Component Of Tongxieyaofang On Visceral Hypersensitivity Related Brain Gut Peptide Of IBS-D

OBJECTIVE: Establishing an IBS-D animal model through colorectal dilatation combined with chronic restraint stress.To observe the influence of the suppressing Mu component and the supportting Tu component of Tongxieyaofang on the expression of brain-derived neurotrophic factor(BDNF),substance P(SP)and calcitonin gene peptide(CGRP)in colon and hippocampus of IBS-D rats.Disscussing the mechanism of the suppressing Mu component of Tongxieyaofang.For the therapy to provide the basis for the experimental mechanism.METHEDS: In this experiment,40 adult male Wistar rats were randomly divided into control group(Con group),model group(Mo group),baishaofangfeng group(B-F group),chenpibaizhu group(C-B group),except for the Con group,the rest rats were modeled by colorectal dilatation(pressure 80 mm Hg)combined with chronic restraint stress(2 hour)for 14 days to construct IBS-D disease and syndrome combined animal model.The criteria for successful modeling are to observe the general condition of the model rat,stool traits,abdominal withdrawal reflex and colorectal pathological examination.and were given traditional Chinese medicine intervention for 14 days the next day.The normal group and the model group were given oral distilled water.The other groups were taken orally with Baishaofangfeng extract(C=0.27g/ml)and Chenpibaizhu(C=0.405g/ml)extract.During the experiment,the general conditions,tweight change,stool characteristics,and the abdominal withdrawal reflex response(AWR)of each group of rats were observed.One week after the treatment,the hippocampus tissue and the colon tissue 6 cm from the end of the rectum were removed.The amount of BDNF、CGRP and SP in the colon and hippocampus of each group of rats were detected by ELISA.RESULT: 1.General situation;Before the establishment of the model,the rats in each group were in good mental state,reacted sensitively,had healthy fur,and had normal bowel movements.After the establishment of the model,the spirit of the rats in the model group showed depression,the fur was disorderly,the food intake and body weight decreased in varying degrees,and feces were sparse.Rats in each treatment group improved in their spirit,mobility,hair luster,diet intake and body weight to varying degrees after medication,and stool gradually formed.2.The weight of the rats;Before the establishment of the model,There was no difference between each groups(p>0.05);After the establishment of the model,the weight gain of rats in the Mo group was significantly lower than the Con group(p<0.05).After the treatment,compared with the Mo group,rats in the B-F group and C-B group grew rapidly(p<0.05).There was no significant difference in weight between the B-F group and the C-B group(p>0.05).3.Stool traits;Before the establishment of the model,there was no significant difference in each group(p>0.05);After the establishment of the model,compared with the Con group,Bristol Stool Scale of the three Mo group were significantly increased(p< 0.05);After the treatment,compared with the Mo group,the Bristol Stool Scale of rats in the B-F group were significantly reduced(p<0.05);there was no difference between the two group,and there was no statistical significance(p>0.05).4.Abdominal withdraw reflex score;After the establishment of the model,the AWR score of the model group was significantly higher than the Con group under the pressure of 20,40,and 60 mm Hg(p<0.05).the difference between the groups disappeared under the pressure of 80 mm Hg.After treatment,compared with the Mo group,the AWR score of rats in B-F group was reduced(p<0.05),and the difference was obviously statistically significant.However,the C-B group had little effect on the reduction of AWR score,and the difference was not statistically significant(p>0.05).5.Pathological examination;The intestinal epithelium of rats in each group was intact and continuous,without edema,ulcer,erosion and inflammatory cell infiltration,which accorded with the pathological changes of IBS-D.6.Animal model evaluation;After chronic restraint stress combined with colorectal dilatation,the rats in the Mon group showed malaise,decreased activity,decreased weight gain(p<0.05),increased the number of excreted stools,and significantly increased the amount of loose stools.The AWR score(p<0.05),the significantly lowered visceral pain threshold,and the clinical features of IBS-D with obvious visceral hypersensitivity and gastrointestinal dysfunction,Combined with colonic pathological changes without inflammatory changes,this method can successfully establish an IBS-D animal model.7.The expression of BDNF in the colon and hippocampus of rats;Compared with the Con group,the expression of BDNF in the colon of the Mo group was significantly increased(P<0.05);the B-F group was significantly lower than the Mo group(P<0.05).The expression of BDNF in the Mo group in the hippocampus was significantly lower than that in the Con group(p<0.05).and the expression of BDNF in the B-F group was significantly higher than that in the Mo group(p<0.05).The C-B group had no significant regulatory effect on the expression of BDNF in the colon and hippocampus.8.SP expression in the colon and hippocampus of rats: The expression of SP in the colon and hippocampus was significantly increased in the Mo group compared with the Con group,and the B-F group was significantly decreased compared with the Mo group(all P<0.05).the C-B group had no significant regulatory effect on the SP expression.9.CGRP expression in rat colon and hippocampus: the same as SP expression.CONCLUSONE: 1.Colorectal distention combined with chronic restraint stress can successfully reproduce Animal Models of IBS-D Disease.In this experiment,the success rate of this experimental method was 85.37%.2.The suppressing Mu component of Tongxieyaofang an significantly improve the symptoms of diarrhea and intestinal hypersensitivity in IBS-D model rats.3.BDNF is positively correlated with the expression of SP and CGRP in the intestinal tract,verifying that BDNF can be used as a modulator to promote the release of SP and CGRP to regulate intestinal function.It has a synergistic effect and jointly participates in the formation of visceral hypersensitivity of IBS-D.4.It may be part of the mechanism of the suppressing Mu component of Tongxieyaofang to treat IBS-D by reducing the release of brain-gut peptides BDNF,SP and CGRP in the colon,reducing inflammatory response and pain information,and alleviating IBS-D visceral hypersensitivity.