| Objective To investigate the therapeutic effect of Xiaoyao powder combined with Bifidobacterium tetravaccine tablets(live)in the intervention of IBS-D rats and to analyze a preliminary mechanism of the brain-gut interaction so as to provide theoretical foundations and data references for its clinical application.Methods A retrospective analysis of relevant literature on IBS-D rat models from both China and abroad was conducted,focusing on the types of experimental animals and modeling methods used in the literature.The IBS-D rat model was established by multifactors were divided into the model group,the Bifidobacterium tetravaccine tablets(live)group(hereafter referred to as the Bifidobacterium group),and the Xiaoyao powder combined with Bifidobacterium tetravaccine tablets(live)group(hereafter referred to as the Xiaoyao and Bifidobacterium group).Additionally,eight rats was used as control group.Rats in the Bifidobacterium group received a Bifidobacterium suspension by gavage at a dose of 0.5g/kg,while those in the Xiaoyao and Bifidobacterium group received a Xiaoyao and Bifidobacterium suspension by gavage at a dose of 10.5g/kg.The model and normal groups given an equal volume of distilled water,all for 14 days.The general behavior,fecal Bristol typing score,and fecal water content,the pressure threshold of visceral sensitivity were measured before and after drug administration for all groups.After treatment,brain and intestinal tissues was observed by HE while goblet cells,the length of intestinal villi,and the depth of crypts of intestinal tissues by AB-PAS for all groups.The contents of SP,VIP,NGF,5-HT,and NPY in rat serum,and 5-HT and NPY in brain and intestinal tissues were measured by ELISA.Expression levels of 5-HTR1 a,5-HTR3 a,and 5-HTR4 Protein in the hippocampus and intestinal tissues were detected by Western blot.Expression levels of 5-HTR1 a,5-HTR3 a,and 5-HTR4 m RNA in intestinal tissues were analyzed by RTq PCR.Results(1)Based on a retrospective analysis,SD rats were the most frequently selected experimental animals for IBS-D and multifactors such as maternal separation.(2)Before treatment,comparing with the control group,general behavioral scores and AWR pressure thresholds reduced significantly(P<0.05)while Bristol scores and fecal water content increased significantly(P<0.05),meanwhile there were no significant differences of the indexes such as goblet cell count,villus length and crypt depth in other groups.SP,VIP,5-HT in serum and 5-HT in brain and intestine increased significantly(P<0.05)while NPY in serum and intestine decreased significantly(P<0.05)in other groups.The expressions of 5-HTR3 a protein and m RNA in brain and intestinal tissues increased significantly(P<0.05)while 5-HTR4 protein and m RNA decreased significantly(P<0.05).(3)After a two-week treatment,comparing with the IBS-D group,Bifidobacterium group were no significant differences of the general behavioral scores and AWR pressure thresholds(P>0.05),while the Bristol scores and fecal water content decreased significantly(P<0.05).SP,VIP,5-HT in serum and 5-HT in intestine decreased significantly(P<0.05)while there were no significant differences of NPY in serum and brain decreased significantly(P<0.05)in other groups.The expressions of 5-HTR3 a protein and m RNA in brain and intestinal tissues decreased significantly(P<0.05)while 5-HTR4 protein and m RNA increased significantly(P<0.05).(4)After a two-week treatment,comparing with the control group,Xiaoyao and Bifidobacterium group were no significant differences of the general behavioral scores(P>0.05),comparing with the IBS-D group,AWR pressure thresholds increased significantly(P>0.05)while the Bristol scores and fecal water content decreased significantly(P<0.05).SP,VIP,5-HT in serum and 5-HT in intestine decreased significantly(P<0.05)while the NPY in brain decreased significantly(P<0.05)in other groups.The expressions of 5-HTR3 a protein and m RNA in intestinal tissues decreased significantly(P<0.05)while 5-HTR4 protein and m RNA increased significantly(P<0.05).Conclusion Utilizing a multifactorial combination of maternal separation,restraint stress,and senna gavage with rectal stimulation by acetic acid could establish an IBS-D rat model,which exhibits clinically relevant symptoms,such as pronounced diarrhea(evidenced by increased Bristol typing scores and elevated fecal water content)and abdominal pain(indicated by increased visceral sensitivity)and no intestinal histopathological changes,simulate the clinical characteristic of IBS-D highly.Both Bifidobacterium tetravaccine tablets(live)and Xiaoyao powder combined with Bifidobacterium tetravaccine tablets(live)could alleviate the symptoms of abdominal pain and diarrhea,combination of Xiaoyao powder and Bifidobacterium tetravaccine tablets(live)shows a superior efficacy.The therapeutic effect of Xiaoyao powder combined with Bifidobacterium tetravaccine tablets(live)may be relation with downregulation of SP,VIP,and 5-HT in serum,up-regulation of NGF and NPY in serum,down-regulation of 5-HT levels in brain and intestinal tissues;up-regulation of NPY levels in intestinal tissues;up-regulation of 5-HTR4 levels in intestinal and hippocampal tissues;and down-regulation of 5-HTR3 a levels in intestinal tissues.Additionally,this therapeutic effect may involve to the down-regulation of 5-HTR3 a m RNA expression in intestinal tissues and the up-regulation of 5-HTR4 m RNA expression in intestinal tissues. |
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