Study On The Regulation Of Banxia Xiexin Decoction On Gastrointestinal Motility Based On Multidimensional Mining And 5-HT₄R/TAS2R38

[Background]Functional gastrointestinal disorders(FGIDs)are a group of digestive disorders with a series of gastrointestinal symptoms such as abdominal distension,nausea,vomiting,abdominal pain,diarrhea,etc.,without obvious organic lesions.Although the pathogenesis of FGIDs is complex,gastrointestinal motility disorder is one of the important pathogenesis.The current treatment of FGIDs is based on pro-gastrointestinal motility drugs,such as 5-hydroxytryptamine agonists,dopamine receptor inhibitors,and gastrin receptor agonists,but there are adverse effects such as cardiac arrhythmias,hyperprolactinemia,and extrapyramidal reactions with long-term administration.Thus,the search for safe and effective treatments is a key issue in the treatment of FGIDs.The Acrid-releaseandbitter-downbearing method,which first originated from the Yellow Emperor’s Classic of Internal Medicine,is one of the important treatments to harmonize the spleen and stomach.As a representative formula of pungent opening and bitter lowering,Ban Xia Xie Xin Decoction from the Treatise on Typhoid has good efficacy both in improving clinical symptoms such as fullness,stomach pain,abdominal distension,belching,acid reflux,vomiting,abdominal pain and diarrhea,and in modern response to FGIDs such as FD,functional diarrhea and IBS.However,most of the current research on thethe Acrid-release and bitter-downbearing method is mainly clinical research,and little research on its molecular mechanism has been reported,which has stimulated a strong desire for research in the subject group!In this study,the group firstly summarized the research dynamics and characteristics of Acrid-release and bitter-downbearing method for the treatment of functional gastrointestinal disorders in the past 5 years;then analyzed the medical cases of Acrid-releaseandbitter-downbearing method for the treatment of FGIDs using multidimensional data technology,and found that the core formula of Acrid-releaseandbitter-downbearing method is Ban Xia Xie Xin Decoction;finally,animal experiments were conducted to study the effects of Ban Xia Xie Xin Decoction on the gastrointestinal dynamics of chronic unpredictable mild stress(CUMS)model mice from the perspective of 5-HT4R/bitter taste receptor(TAS2R38).Finally,we investigated the effects of Ban Xia Xie Xin Decoction on the gastrointestinal motility of mice in the chronic unpredictable mild stress(CUMS)model.This study not only summarized the rules of treatment of FGIDs by the Acrid-release and bitter-downbearing methods,but also explored the synergistic mechanism of the Acrid-release and bitter-downbearing methods in FGIDs represented by Ban Xia Xie Xin Decoction,which provides a model for guiding the complex mechanism of action of multi-component and multi-target of Chinese herbal compound,with important scientific significance and application value.[Research methods]1.Overview of literature-based studies on the treatment of functional gastrointestinal disorders with the Acrid-release and bitter-downbearing methodIn this study,firstly,literature search was conducted by searching China Knowledge Network,Wanfang database,Vipshop database,and China Biomedical Literature Database;secondly,literature management was performed on all downloaded literature using EndNoteX9.Next,duplicate literature was firstly excluded and excluded by reading the title abstract of the literature that did not meet the inclusion criteria of this study,and literature that did not match the content of this study was excluded by checking the weight and reading the title,abstract or full text.Finally,bibliometric analysis was used to sort out and summarize the research dynamics and characteristics of the last 5 years of Acrid-release and bitter-downbearing method for the treatment of functional gastrointestinal diseases for the final screening literature.2.A retrospective study of medical cases of functional gastrointestinal diseases based on multidimensional data mining of the Acrid-release and bitter-downbearing methodThe medical cases of functional gastrointestinal disease treated with the Acrid-releaseandbitter-downbearing method were searched from CNKI,VIP,Wan Fang Data Knowledge Service Platform(new version),and China Biomedical Information Database(CBID)from the time of database construction to December 2021.The core search terms were"irritable bowel" "functional" "reflux disease" " hysteria" "belching" "gastric distention""stomach pain" "acid reflux" "diarrhea" "constipation" "idiopathic nausea" "periodic vomiting" "regurgitation" "sphincter dysfunction" " functional gastrointestinal disorders"were searched by the subject terms "Xin Kai Bitter Lowering" "Xin Tong Bitter Lowering""Banxia Xiexin Decoction","Zuo Jin Wan",and The search results were incorporated into the EndNote document management system for checking,screening the documents,reading the full text,extracting the medical cases that met the criteria,and entering the evidence,pathogenesis,prescriptions,tongue and pulse into MedcaseV5.2."MedcaseV5.2 Chinese medicine practitioner workshop"-famous old Chinese medicine practitioner experience inheritance support platform,and establish a database.The data of 246 initial consultations that met the inclusion criteria were analyzed by data spectrum analysis,data association rules,data clustering analysis,object dispersion clustering,prescription dispersion clustering,drug cohesion layer clustering and other aspects of the experience summary from the tongue,pulse,disease mechanism,prescription medication,etc.,to explore the symptoms,evidence,tongue and pulse,prescription medication and other contents of the medical cases,in order to explore the pungent opening and bitter lowering method of treating functional gastrointestinal diseases.The clinical rules of medication.3.Study on the regulation of gastrointestinal motility by Ban Xia Xie Xin Decoction based on 5-HT4R/TAS2R38We used the chronic stress composite modeling method to construct CUMS mice,and observed the effects of Ban Xia Xie Xin Decoction and its dismantled formula on body weight,food intake,hair color and general condition,as well as gastric emptying and small intestine propulsion rate of the model mice.The effect of different concentrations of Ban Xia Xie Xin Decoction and its dismantled formula on the contraction of smooth muscle strips in normal SD rats was evaluated by in vitro intestinal tube assay;the effect of Ban Xia Xie Xin Decoctionand its dismantled formula on ChAT protein,5-HT4R,TAS2R38 and its downstream protein(p-ERK)in model mice was observed by immunoprotein blotting and immunofluorescence techniques.[Research results].Overview of studies based on the literature on the treatment of functional gastrointestinal disease by the Acrid-release and bitter-downbearing methodA total of 25 papers were included in the literature.In terms of study type,clinical studies accounted for 21 articles,while experimental studies were only 4.In terms of researchers,4 master’s theses were included;in terms of study diseases,12 articles were on functional dyspepsia,3 articles were on irritable bowel syndrome,4 articles were on gastric gangrene,1 article was on gastroparesis,1 article was on functional constipation and 1 article was on chronic diarrhea;the study mechanisms were mainly on gastrointestinal dynamics and brain and intestinal peptides.2.A retrospective study of medical cases of functional gastrointestinal diseases based on multidimensional data mining of the Acrid-releaseandbitter-downbearing methodThe medical cases eligible for enrollment were 132 cases with 246 consultations,and the specific findings were as follows.2.1 The frequency distribution of gender and age,the distribution of men and women in patients differed significantly,the ratio of men to women was 5:3,and there were significantly more male patients than female.2.2 In terms of frequency of pathogenesis,the highest frequency distribution of pathogenesis of functional gastrointestinal disease(gastric gangrene)was cold-heat mismatch,stomach loss of harmony and descent;the highest frequency distribution of treatment rules was chemistry and stomach,Acrid-release and bitter-downbearing.2.3 Among the top 30 drugs in the drug frequency table,the highest frequency distribution of drugs was 6.97%,6.81%,and 6.03%for Han Xia,Huang Lian,and Gan Jiang,respectively.2.4 Drug K-means clustering 12 clusters,pungent open bitter down,strengthen the spleen and stomach as the main clusters,but the emergence of pairs of drugs,corner drug clusters,such as Huanglian,dry ginger cold and heat leveling regulation,Panax quinquefolium,Scutellaria baicalensis clear heat and diarrhea of the liver,dispersing cold to stop vomiting.There are also 4-bit combinations:such as dry ginger,half summer,scutellaria,Huanglian and other cluster drugs,suggesting that the drug cluster may have a cluster effect of Acrid-release and bitter-downbearing,lowering rebellion and stopping vomiting.2.5 Among the 12 categories of prescriptions clustered by K-means,according to the principle that the core value of prescription clustering is greater than the median value[R:N>M]and the principle of clustering,the core prescriptions were finally excavated as:Ban Xia,Huang Lian,Gan Jiang,Licorice,Scutellaria and Hou Pu.This is known as Ban Xia Xie Xin Decoction.3.Study on the modulating effect of Ban Xia Xie Xin Decoction on gastrointestinal motility based on 5-HT4R/TAS2R383.1 Compared with CUMS model group,Ban Xia Xie Xin Decoction group and Acrid-release and bitter-downbearing group could improve the gastric emptying rate and small intestinal propulsion rate of CUMS model mice,and the difference was statistically significant(P<0.05);Compared with the whole formula group,the whole formula group of Ban Xia Xie Xin Decoction increased the intestinal propulsion rate of CUMS model mice,which was better than that of bitter-downbearing group alone,and the difference was statistically significant(P<0.01);Compared with the CUMS model group,Ban Xia Xie Xin Decoction group and Acrid-release and bitter-downbearing group could increase the secretion of 5-HT,MTL,Ghrelin,GAS,GLP-1,LEP and other gastrointestinal hormones in CUMS model mice,except CCK,and the difference was statistically significant(P<0.05).Compared with the whole formula group,Ban Xia Xie Xin Decoction group promoted the secretion of 5-HT and MTL in CUMS model mice better than Acrid-release and bitter-downbearinggroup The difference between Acrid-release and bitter-downbearingg roup was statistically significant(P<0.01);In the normal mouse model,compared with the blank control group,there was no significant difference in gastric emptying rate and small intestinal propulsion rate in each treatment group of Banxia Xiexin Decoction(P>0.05).Compared with the blank control group,Acrid-release and bitter-downbearing group of Ban Xia Xie Xin Decoction could promote MTL secretion,and there was a significant difference(P<0.05);Banxia Xiexin Decoction had no significant effect on the secretion of 5-HT,MTL,ghrelin,GLP-1 and CCK.3.2 compared with the blank group,① Ban Xia Xie Xin Decoction Acrid-release group had no significant effect on the contraction inhibition rate of duodenal smooth muscle under any concentration.② At the normal equivalent concentration,the Ban Xia Xie Xin Decoction group had no significant effect on the contraction inhibition rate of duodenal smooth muscle.Only when the administration concentration was 1.3mg/ml,the Ban Xia Xie Xin Decoction group increased the inhibition rate of duodenal smooth muscle contraction.③ When the administration concentration was 0.8mg/ml,the inhibitory rate of Ban Xia Xie Xin Decoction Acrid-release and bitter-downbearing group on duodenal smooth muscle contraction increased,which was related to the change of concentration.④ The contraction rate of duodenal smooth muscle increased with the concentration of Ban Xia Xie Xin Decoction(0.6mg/ml),and the contraction rate of duodenal smooth muscle increased gradually in the Ban Xia Xie Xin Decoction group.3.3 compared with CUMS model group,Ban Xia Xie Xin Decoction group,Ban Xia Xie Xin Decoction Acrid-release and bitter-downbearing group and Ban Xia Xie Xin Decoction Acrid-release group could increase the protein expression of chat in gastric antrum and small intestine,and the difference was statistically significant(P<0.01).Compared with Quanfang group or Acrid-release and bitter-downbearing group,Ban Xia Xie Xin Decoction group or Acrid-release and bitter-downbearing group increased the expression of chat in small intestine,which was better than Acrid-release group and bitter-downbearing group alone(P<0.05,P<0.01).Ban Xia Xie Xin Decoction group or Acrid-release and bitter-downbearing group was better than bitter-downbearing group alone(P<0.05,P<0.05).3.4 compared with the CUMS model group,except the bitter-downbearing group,the other treatment groups of Ban Xia Xie Xin Decoction can increase the expression of 5-HT4R protein in small intestine of CUMS model mice,and the difference is statistically significant(P<0.01,P<0.05).Ban Xia Xie Xin Decoctionn group and Acrid-release and bitter-downbearingcan promote the expression of 5-HT4R protein in gastric antrum of CUMS model mice,and the difference is statistically significant(P<0.01,P<0.001);Compared with Quanfang group or Acrid-release and bitter-downbearing group,the expression of 5-HT4R protein in gastric antrum of CUMS model mice in Ban Xia Xie Xin Decoction group and Acrid-release and bitter-downbearing group were better than Acrid-release group and bitter-downbearing group,and the difference was statistically significant(P<0.01,P<0.001);Immunohistochemical results showed that compared with the model group,except the bitter-downbearing group,the other treatment groups of Ban Xia Xie Xin Decoction increased the expression of gastrointestinal 5-HT4R inCUMS model mice,and the difference was statistically significant(P<0.001,P<0.01,P<0.05).Compared with the Quanfang group,the increase of gastrointestinal 5-ht4r expression in Ban Xia Xie Xin Decoction Quanfang group was better than that in Acrid-release and bitter-downbearinggroup,Acrid-release group and bitter-downbearing group,and the difference was statistically significant(P<0.01,P<0.05);The results of immunofluorescence showed that compared with the CUMS model group,the expression level of 5-HT4R in gastrointestinal tract(gastric antrum,small intestine and colon)of CUMS model mice in Ban Xia Xie Xin Decoction group was higher and had statistical difference(P<0.0001).Compared with the whole formula group or Acrid-release and bitter-downbearing group,the expression of 5-HT4R in gastrointestinal tract(gastric antrum and small intestine)of CUMS model mice in Ban Xia Xie Xin Decoction group andAcrid-release and bitter-downbearing group was better than that in bitter-downbearing group(P<0.05),Acrid-release group increased the expression of 5-HT4R in the small intestine of CUMS mouse model,which was better than that of bitter-downbearing group(P<0.05).3.5 compared with CUMS model group,Ban Xia Xie Xin Decoction group and bitter-downbearing group can promote the expression level of TAS2R38 in gastrointestinal tract of CUMS model mice,and the difference is statistically significant(P<0.05).Ban Xia Xie Xin Decoction Acrid-release and bitter-downbearinggroup can increase the expression level of TAS2R38 in small intestine of CUMS model mice,and the difference is statistically significant compared with Acrid-release group,Ban Xia Xie Xin Decoction bitter-downbearing group increased the expression of TAS2R38 in the small intestine of CUMS model mice,which was better than Acrid-release group,and the difference was statistically significant(P<0.01);Immunohistochemical results showed that compared with CUMS model group,except Acrid-release group,the expression of TAS2R38 in gastrointestinal tract of CUMS model mice in other groups of Ban Xia Xie Xin Decoction group was significantly increased(P<0.05),The expression level of TAS2R38 in gastric antrum of CUMS model mice in Ban Xia Xie Xin Decoction group and Acrid-release bitter-downbearing group was significantly higher than that in Acrid-release group and bitter-downbearing group alone(P<0.0001).The expression of TAS2R38 in intestinal tract of CUMS model mice in Ban Xia Xie Xin Decoction group and Acrid-release bitter-downbearing group was significantly higher than that in Acrid-release group alone(P<0.05);Compared with the CUMS model group,except the Acrid-release group,the other treatment groups of Banxia Xiexin Decoction could increase the expression level of p-ERK in the gastrointestinal tissues(antrum,small intestine and colon)of CUMS model mice,and the difference was statistically significant(P<0.05).Compared with the Quanfang group,the Ban Xia Xie Xin Decoction Quanfang group increased the expression of p-ERK in the gastrointestinal tissues(antrum and small intestine)of CUMS model mice,which was better than the Acrid-release roup alone.Compared with theAcrid-release and bitter-downbearing group,The increase of p-ERK expression in intestinal tissue(small intestine and colon)of CUMS model mice in Ban Xia Xie Xin Decoction Acrid-release and bitter-downbearing group was better than that in Acrid-release group alone,and the difference was statistically significant(P<0.05).[ConclusionandSignificance]1.Through the literature research in the past 5 years,it was found that most of the case data of Acrid-releaseandbitter-downbearing in the treatment of functional gastrointestinal diseases were reported on a case-by-case basis,or only summarized a certain disease type in functional gastrointestinal diseases,and did not summarize and analyze the data of all functional gastrointestinal disease case studies,and did not systematically organize the clinical use pattern of Acrid-releaseandbitter-downbearing method in functional gastrointestinal diseases,and did not see the literature report of mining the clinical use pattern of Acrid-releaseandbitter-downbearing method in functional gastrointestinal diseases based on multidimensional data.Therefore,the present study was based on multidimensional data.Therefore,the present study is of great scientific significance in the study of the rules of the diagnosis and treatment of functional gastrointestinal diseases based on the multidimensional data mining of theAcrid-release and bitter-downbearing method.2.Based on the multidimensional data mining analysis,the main pathogenesis of gastroparesis is loss of harmony and descent of the stomach,dampness and heat,and weakness of the spleen and stomach,and the core drugs are mainly the internal correlation item set among six drugs,namely,Radix Codonopsis pilosulae,Radix Scutellariae,Radix Scutellariae,Glycyrrhiza glabra,Radix Panax notoginseng,and Radix Ginger.The two drugs are not only a famous medicine pair for lowering rebellion and stopping vomiting,but also one of the representative medicine pairs of Acrid-release and bitter-downbearing,thus it can be seen that the key treatment method of Acrid-releaseandbitter-downbearing for functional gastrointestinal diseases is the core formula of Ban Xia Xie Xin Decoction,and the core medicine pairs are Huang Lian and Ban Xia.This data mining can provide an important theoretical basis for the subsequent in-depth exploration of the synergistic molecular mechanism of the Acrid-releaseandbitter-downbearing Method by summarizing the laws related to the Acrid-releaseandbitter-downbearing Method in the treatment of functional gastrointestinal diseases.3.Ban Xia Xie Xin Decoction increased gastrointestinal motility in CUMS model mice.Acrid-release group and bitter-downbearing group regulated intestinal nerve movement and promoted gastrointestinal motility by affecting the expression of 5-HT4R and TAS2R38 protein respectively.This experiment provides a scientific basis for clarifying the mechanism of multi-component and multi-target biological synergistic effect of traditional Chinese medicine,and is conducive to revealing the scientific connotation of Acrid-release bitter drop method.