Effects Of Chenpi Huoxiang Decoction In Diarrhea Rats Based On Metabolomics And Intestinal Microecology

Objective:Chenpi Huoxiang Decoction is derived from the classic medical book"Yixue Congzong Lu",which is used to treat cholera,vomiting and diarrhea.However,its pharmacological mechanism have not been reported in experimental studies.In addition,Chenpi Huoxiang Decoction is a classic prescription for the compatible application of Pericarpium Citri Reticulatae(Chenpi)and Pogostemonis Herba(Guanghuoxiang).They have the effect of relieving vomiting and diarrhea as the herb pair,and have a wide range of clinical applications(including 24 prescriptions of ChP.2015 and 78 prescriptions of"Chinese traditional patent medicines"),but lack of modern medical evidence to clarify the scientific connotation of the compatibility.Therefore,based on metabolomics and intestinal microecology,this study aimed to explore the efficacy and related pharmacological mechanism of Chenpi Huoxiang Decoction,and to clarify the scientific connotation of the compatibility of Chenpi and Guanghuoxiang.Methods:(1)The diarrhea rat models were established by oral administration of folium sennae decoction.After successful modeling,they were treated with Chenpi Huoxiang Decoction,Chenpi single decoction and Guanghuoxiang single decoction,respectively.The diarrhea index,the weight gain rate and the intestinal ink propellation rate of rats were evaluated for their therapeutic effects.(2)To characterize the untargeted metabonomics profiles in the rats,we analyzed blood,urinary and fecal metabolites by ~1H NMR technique.then the NMR data were analyzed by multivariate statistical analysis and univariate statistical analysis.The differential metabolites associated with the model and the therapeutic drug were screen out,and were used to identify the affected metabolic pathways.(3)To characterize the fecal microbiota communities of the rats,we performed high throughput sequencing analysis in region 4 of bacterial 16S rDNA through the Illumina Hiseq2500 platform.Then the bioinformatics analysis of the sequencing data was performed on the QIIME platform.PICRUSt was used to predict the metabolic function of gut microbiota.The effects of the gut microbiota on host metabolism were analyzed in conjunction with metabolomics results.Results:(1)After modeling,the rats discharged loose stools,and their weight growth stagnated or even decreased.After administration,the diarrhea index of the three administration groups was significantly decreased.Their body weight growth rate was higher than that of recovery group.Like the Chenpi Huoxiang Decoction group,the difference between the Chenpi group and the recovery group was significant.Furthermore,the intestinal ink propelling rate of the Chenpi Huoxiang Decoction group was significantly increased,which was close to the normal level.(2)After modeling,23 urinary metabolites and 18 fecal metabolites were selected as the differential metabolites;4 metabolic pathways were disturbed,including Valine,leucine and isoleucine biosynthesis,Citrate cycle,Glyoxylate and dicarboxylate metabolism,Glycine,serine and threonine metabolism.After treatment,Chenpi Huoxiang Decoction significantly altered 11 serum metabolites,16 urinary metabolites and 10 fecal metabolites,so that most of them were close to normal levels.Chenpi single decoction significantly changed 12 serum metabolites,12 urinary metabolites and 6 fecal metabolites.Guanghuoxiang single decoction significantly modulated 10 serum metabolites,6 urinary metabolites,and 11 fecal metabolites.According to the results of three biological samples,we found that the effect of Chenpi Huoxiang Decoction on metabolism of diarrhea rats was better than that of Chenpi single decoction and Guanghuoxiang single decoction,which affected Butanoate metabolism,Citrate cycle,Alanine,aspartate and glutamate metabolism,Histidine metabolism,Glyoxylate and dicarboxylate metabolism,Synthesis and degradation of ketone bodies,D-Glutamine and D-glutamate metabolism.(3)After modeling,diversity of gut microbiota was decreased in diarrhea rats.and the gut microbiota characterized by a higher level of opportunistic pathogens and a lower level of butyrate-producing bacteria;PICRUSt analysis showed that the metabolic functions of gut microbiota were significantly changed,including pyruvate,citrate,butanoate,methane and tryptophan.After administration,like the Chenpi Huoxiang Decoction group,diversity of gut microbiota in the Guanghuoxiang group was lower than that of the recovery group;The gut microbiota structure of the Chenpi Huoxiang Decoction group and the control group was the most similar,indicating that Chenpi Huoxiang Decoction can promote Prevotella,Allobaculum and Blautia and inhibit the growth of Bacteroides,Escherichia and Clostridium,making the composition of gut microbiota closer to the control group than the Chenpi group and the Guanghuoxiang group.The comprehensive data analysis implied that Chenpi Huoxiang Decoction modulated the metabolic function(pyruvate,butanoate,histidine and bile acid)of gut microbiota to improve host metabolism.Conclusion:Chenpi Huoxiang Decoction has obvious intervention effect on metabolism and gut microbiota of diarrhea rats,and can improve host metabolism by regulating gut microbiota.The compatibility of Chenpi and Guanghuoxiang has a synergistic effects on the treatment of diarrhea.