Based On The "intestinal Mucosal Immune Barrier-intestinal Flora" The Mechanism Of Action Of Traditional Chinese Medicine In The Treatment Of IBS-D

BACKGROUND: Irritable bowel syndrome?IBS?is a functional intestinal disease characterized by recurrent abdominal pain,which is associated with defecation or acc ompanied by changes in defecation habits.The global prevalence of IBS ranges from 7% to 21%.In China,the prevalence of IBS ranges from 0.82% to 5.67% according t o the same diagnostic criteria.IBS symptoms often recur.In addition to gastrointestinal symptoms,parenteral symptoms such as palpitation,insomnia,dizziness,headache and fatigue can seriously affect the quality of life of patients.According to defecation habit s,IBS can be divided into the following subtypes: irritable bowel syndrome with pred ominant constipation?IBS-C?,irritable bowel syndrome with predominant diarrhea?IBS-D?,and irritable bowel syndrome with mixed bowel habits?IBS-M?.Among them,IB S-D has the highest prevalence.For IBS patients,IBS-D-related symptoms are the mai n factors leading to patients seeking treatment in medical institutions.The clinical sym ptoms of IBS-D were fecal frequency increasing and fecal sparseness.The type of sto ol accorded with type 6 and 7 of Bristol Fecal Traits Scale.The pathogenesis of IBS is not yet clear.Current studies on IBS mainly focus on disorder of gut-brain interacti on.In the past,the understanding of functional gastrointestinal disorders?FGIDS?sym ptoms was mainly dynamic abnormality.With the development of medicine,the curren t understanding has turned to Neurogastroenterology and brain-intestinal interaction and other abnormalities.At present,IBS symptoms are considered to be mainly caused by the following aspects: dynamic disorders,visceral hypersensitivity,changes in mucosal and immune functions,changes in intestinal flora,abnormal treatment of the central ne rvous system?CNS?.Among them,the characteristic changes of intestinal flora and int estinal mucosal barrier are the focus of IBS research.It is also the target of IBS treat ment.Modern medical treatment mainly includes lifestyle adjustment,microecology/im munomodulator,drug treatment for diarrhea,constipation,abdominal pain,and anti-anxi ety and depression treatment.Traditional Chinese medicine has accumulated a lot of e xperience in the treatment of this disease.IBS-D includes abdominal pain and diarrhea,which belong to the category of "diarrhea" and "abdominal pain",in which stagnation of liver qi and spleen deficiency are the core pathogenesis of the disease.Soothing th e liver and regulating the spleen is the key to the treatment of IBS-D.Modern Chinese medicine research shows that traditional Chinese medicine compound can treat the dis ease with multiple targets,such as improving intestinal immune inflammatory response,regulating abnormal brain-intestinal interaction,improving intestinal flora,etc.OBJECTION: To discuss the pathogenesis of IBS-D and to elucidate the material basis and mechanism of TCM compound Tongxieyaofang Decoction in the treatment of IBS-D.METHOD: In this study,human trials are used,which are divided into three parts.The first part is the evaluation of clinical efficacy.A total of 34 patients with IBS-D were enrolled.They were randomly divided into two groups: the compound Chinese medicine group?IBS-d,n=24?and the blank control group?IBS-dCG,n=10?.At thesame time,healthy volunteers were recruited as healthy control group?Health,n= 30?.The basic information of the participants was registered and the case report form was recorded.The traditional Chinese medicine compound group was given Tongxieyaofang Decoction for four weeks,while the blank control group was given general non-drug treatment for four weeks.HAD,Wong-Baker score and TCM symptom score before and after treatment were taken as outcome indicators.To observe the curative effect of traditional Chinese medicine compound on IBS-D.The second part is the study of intestinal mucosal barrier.A total of 26 IBS-D patients with liver-depression and spleen-deficiency syndrome were included,while The healthy control group included 20 patients.10 of 26 IBS-D patients were selected by random number table method for follow-up.Blood DAO,D-LA,IL-1?,IL-10 and 5-HT were detected by ELISA in healthy group and IBSD patients before and after treatment with traditional Chinesemedicine compound.By comparing the changes of plasma parameters in different groups,the pathogenesis of IBS-D patients and the effect of traditional Chinese medicine compound on intestinal mucosal barrier were clarified.The third part is the study of intestinal microorganisms.A total of 30 IBS-D patients with liver depression and spleen deficiency were included in this study.A total of 20 healthy controls were recruited.20 IBS-D patients were selected from 30 IBS-D patients by random number table method and randomly divided into two groups.One group was Chinese medicine compound group?n=10?.The other group was blank control group?n=10?.The changes of intestinal flora in healthy group and IBS-D patients were detected by 16 S DNA method before and after treatment with Chinese medicine compound.To explore the pathogenesis of IBS-D from intestinal microorganisms and the intervention target of Chinese medicine compound.RESULTS: Part I clinical trial: thirty-four patients with IBS-D were included,and one patient was excluded due to unqualified specimen collection.All the remaining patients completed the trial successfully.The difference in HAD scores between the healthy control group and IBS-D patients was statistically significant?P<0.05?,and the HAD score of IBS-D patients was significantly higher than that of the healthy control group,indicating that anxiety and depression were prevalent in IBS-D patients.The HAD score of the traditional Chinese medicine compound group after the treatment ofTongxieyaofang Decoction was not statistically significant compared with that before the treatment?P>0.05?.The difference of Wong-Baker scores before and after treatment inthe compound medicine group was statistically significant?P<0.05?,indicating that the Chinese herbal compound has an obvious effect on alleviating patients' abdominal pain symptoms.The difference of TCM syndrome points before and after treatment was statistically significant?P<0.01?.In IBS-D patients,the symptoms of abdominal pain?P<0.05?,loose stool?P<0.05?,Borborygmus andfarting?P<0.05?,abdominal pain for diarrhea?P<0.05?,diarrhea urgency?P<0.05?,shortness of urine?P<0.05?,and anal burning?P<0.05?showed statistically significant remission before and after treatment.In addition,all other symptoms were relieved after the intervention of traditional Chinese medicine compound,but the degree of relief was not significant,and there was no statistical significance.After treatment,2 cases were cured,2 cases were markedly effective,18 cases were effective and 2 cases were ineffective.The total effective rate of Chinese medicine compound group was 91.66%.Blank control group,before and after the treatment of traditional Chinese medicine symptoms integral difference was statistically significant?P<0.05?.After general non-drug treatment,0 cases were cured,0 cases were markedly effective,5 cases were effective,and the total effective rate was 50%.General non-drug treatment can improve the symptoms of patients with IBS-D,but the overall effective rate is not high,the effect is not significant.Comparison between the two groups after treatment of traditional Chinese medicine compound group and blank control group,Wong-Baker has significant difference?P<0.01?,Chinese medicine syndrome integral has significant difference?P<0.01?,HAD no significant difference?P>0.05?,the above results show: tongxieyaofang flavored treatment of IBS-D has good efficacy.It can significantly improve the symptoms of patients with IBS-D.Especially to improve the symptoms of abdominal pain,runny stool,bowel sounding,abdominal pain diarrhea,urgent diarrhea,short and red urine,anal burning and so on.However,the four-week treatment of traditional Chinese medicine compound can not improve the anxiety and depression symptoms of patients.The second part is the results of intestinal mucosal barrier test.Among the 10 patients who received the intervention of TCM,one patient dropped out of the experiment,the rest of the patients completed successfully,and no adverse reactions occurred during the experiment.Compared with IBS-D patients and healthy people,plasma levels of D-LA,DAO,IL-1beta,IL-10,5-HT in IBS-D patients were significantly higher than those in healthy people?P<0.01?,while plasma levels of IL-10 were significantlylower than those in healthy people?P<0.01?.Four weeks after treatment with TCM compound granules,D-LA,DAO,IL-1beta,5-HT were significantly lower than before treatment?P<0.01?,but still higher than the level of healthy people.IL-10,was significantly higher than before treatment?P<0.01?,but still lower than the level of healthy people.The results of this part showed that TCM compound could reduce the intestinalmucosal permeability of IBS-D patients and improve the clinical symptoms of IBS-D by lowering the levels of proinflammatory cytokines and plasma brain-gut peptide,andat the same time increasing anti-inflammatory cytokines.The third part is the results of intestinal flora test: Among the 30 patients with I BS-D included in this study,1 patient was excluded due to unqualified fecal specimenc ollection.All other subjects completed the test successfully.16 SrDNA was used to dete ct the intestinal flora of participants.The results showed that there were differences in flora structure between healthy group and IBS-D patients.Bacteroidetes,Actinobacteria a re a phylum level differential species?P<0.05?;Bacteroidia,Actinobacteria are class lev el differential species?P<0.05?;Bacteroidales,Bifidobacteriales,Pasteurellales are order l evel differential species?P<0.05?;Peptostreptococcacea,Bifidobacteriaceae,[Paraprevotel laceae],Enterococcaceae,Pasteurellaceae are family level differential species?P<0.05?;Haemophilus,Enterococcus,Paraprevotella,Bifidobacterium,Faecalibacteriumare genus leve l differential species?P<0.05?.There was no significant differencein Alpha diversity be tween the healthy group and the IBS-D group?P>0.05?.The Betadiversity analysis sh owed that the composition of microflora of the two groups wassignificantly different.LefSe analysis showed that there were several species in IBS-D group which LDA sc ore>2: Such as paraprevotellaceae,symergistaceae,S24₇,synergistaceae.Among them B acteroidetes,Bacteroidia,Bacteroidales LDA score>4,Is the most influentialmicroflora am ong the species with significant differences in IBS-D patients.In the healthy group,theLDA score>2 biomarkers of the different species were:enterococcus,enterococcaceae,po rphyromonas,porphyromonas,bacillales,dehalobacterium,dehalobacteriaceae,SMB53,Com amonadaceae,Comamonas,Catenibacterium,bacillaceae.In the healthy group,the LDA score>3 biomarkers of the different species were:Peptostreptococcaceae,Bifidobacterium,Actinobacteria,Bifidobacteriales,Bifidobacteriaceae,Faecalibacterium.Faecalibacterium is the most influential bacterium in the significantly different species in the healthy grou p.In the TCM compound group,Firmicutes and Bacteroidetesranked first and second i n relative abundance at different classification levels,respectively.The ratios before and after treatment were respectively?53.4%vs 48.1%?,?43.2%vs 45.4%?;Clostridia,Bac teroidia?52.5% vs 47.0%?,?43.2% vs 45.4%?;Clostridiales,Bacteroidales?52.5% vs47.0%?,?43.2% vs 45.4%?;Lachnospiraceae,Bacteroidaceae?34.3% vs 25.5%?,?23.8% vs 26.6%?,Bacteroides,Prevotella?23.8% vs 26.6%?,?18.6% vs 17.2%?,After f our weeks of intervention by Chinese herbal compound,relative abundance values of Bacteroidetes,Bacteroidia,Bacteroidales,Bacteroidaceae,Bacteroides were increased com pared with those before treatment.However,Firmicutes,Clostridia,Clostridiales,Lachn ospiraceae,and Prevotella have decreased relative abundance values,but no statistical significance.After the treatment of IBS-D with traditional Chinese medicine,the struct ure of intestinal florachanged,and the phylum level TM7 was the main differential sp ecies?P<0.05?,while genus level Mogibacterium is the main difference species?P<0.05?.LefSe analysis did not find any difference between the two groups of species.Com pared with the blank control group,there was no completely consistent change in inte stinal bacteria,indicatingthat the traditional Chinese medicine compound was suppleme nted with intestinal bacteria such as TM7 as the target,and then affected the intestina l mucosal barrier to achieve the therapeutic effect.CONCLUSION: The pathogenesis of IBS-D is related to intestinal mucosal immu ne disorder,intestinal flora disorder and neuroendocrine imbalance.The above three as pects together lead to the increase of intestinal mucosal permeability,and the increase of intestinal mucosal permeability is more likely to lead to the contact of intestinal an tigen substances with intestinal mucosal antigen presenting cells,thus stimulating more inflammatory stimulators and aggravating intestinal mucosal immunity.Damage to the epidemic barrier.That is to say,the increase of intestinal mucosal permeability is caus al to the damage of intestinal mucosal immune barrier,while Tongxieyaofang Decoctio n can alleviate the symptoms of patients.The mechanism of its therapeutic effect maybe related to the regulation of intestinal immunity,intestinal microecology and intestina l mucosal permeability.