Investigation On Clinical Efficacy,gut Microbiota And Urine Metabonomics Of Gui Fu Di Huang Pill In Treating Yang-Deficiency Constitution

BackgroundYang-deficiency constitution is a dominant part of bias constitution.Guifu Di huang Pill is commonly used to treat yang-deficiency constitution but the mechanism has not been fully understood.The gut microbiota is relevant to kinds of diseases,Urine metabonomics can also reflect the status of the body.Therefore,a randomized clinical trial with gut microbiota study and urine metabonomics to understand the characteristic of yang-deficiency constitution,and the effectiveness and mechanism of Guifu Dihuang Pill in treating yang-deficiency constitution-ObjectiveTo evaluate the effect and mechanism of Guifu Dihuang Pill in treating yang-deficiency constitution.Methods1.Clinical trial62 participants were involved and randomly divided into lifestyle intervention control group(n=31)and Gui fu di huang pill experimental group(n=31).TCM constitution was determined according to the conversion scores.2.Gut microbiota studySubjects were chosen randomly from the clinical trial,16 balanced constitution subjects also were involved in the project.Samples were collected and 16s rRNA method were applied.Parallel-meta 3 is applied for data analysis.3.Urine metabonomics analysisSubjects were chosen randomly from the clinical trial.Urine were colleted for 1H NMR analysis.Metaboanalyst is applied for multivariate statistical analysis to find significant metabolites,enrichment process and pathway.4.Integrated analysis and statistic methodsWGCNA was applied to integrated analyzed the relationship between microbiota genus and metabolites.SPSS 20.0 was used for statistic analysis.Chi-square test,independent-samples t test was used.P<0.05 was considered significant.Results1.Clinical trials of Guifu Dihuang pill in treating yang-deficiency constitution.Conversion scores of yang-deficiency decreased at 12 months after treatment in control group(P<0.05).conversion scores and symptoms of yang-deficiency constitution in the experimental group decreased at all time nodes after intervention(P<0.05).No adverse events happened.2.The gut microbiota research of Gui fu di huang pill in intervening yang-deficiency constitutionBacteroidetes go higher than Firmicutes in yang-deficincy.Bac_Bacteroides and Ery_Asteroleplasm increased in yang-deficiency(P<0.05).Many gut microbiota decreased in yang-deficiency subjects as Rum Ruminococcus,Ent_Enterobacter,Vei Megamonas,et al(P<0.05).Gut microbiota constitution diversity and clustering become closer to balanced constitution status after intervention.3.Urine metabolomic research of Gui fu di huang pill in intervening yang-deficiency constitution70 potential different metabolites were found in yang-deficiency and 57 potential different metabolites were found after drug intervention.These metabolin were mainly enriched and related to kinds of biological processes and pathways.4.Integrated analysis for microbiota,urine metabolites and cytokinesKinds of special bacteroidetes are negative correlated with 70 metabolites in yang-deficiency constitution.Conclusions1.Guifu Dihuang Pill is efficient in treating yang-deficiency constitution in a short time and lasts for a long time.2.The gut microbiota diversity of yang-deficiency constitution was significantly decreased.Gut microbiota diversity recovered to a certain extent in experiment group after intervention with Guifu Dihuang Pill.Potential different metabolites were found between yang-deficiency constitution subjects and balanced cases.All these metabolites were involved and enriched in kinds of biological processes and pathways.3.Dysbacteriosis and metabolic disorders happened in yang-deficiency constitution.Guifu Dihuang Pill may regulate through these ways.