Study Of Jianpi Mixture On Intestinal Microbiota Of Diarrhea Irritable Bowel Syndrome With Spleen Deficiency

Objective:To explore the effects of Jianpi Mixture on intestinal microbiota’ changes of Diarrhea irritable bowel syndrome(IBS-D)with spleen deficiency based on 16S rDNA sequencing analysis,and observe its clinical efficacy.Methods:1.Experimental research:48 young ICR male mice were randomly divided into four groups(n=12),and establish a mouse model of spleen deficiency type IBS-D(3n=36).After successful model evaluation,the blank group and the spleen-deficiency type IBS-D group were given double steam(10ml/kg)gavage,the compound lactic acid bacteria group was given compound lactic acid bacteria gavage,and the Jianpi mixture group was given Jianpi mixture(16ml/kg)intragastrically gavage.The treatment cycle was 14 days.At the end of the treatment period,5 mice were randomly selected from each group,and then the changes of intestinal microbiota in the mice before and after treatment were analyzed by 16S rDNA high-throughput gene sequencing.2.Clinical research:In this experiment,20 healthy volunteers were selected as the blank control group(N),and 23 patients with IBS-D of spleen deficiency type were selected as the treatment group,including 5 in the western medicine group(WM)and 18 in the Chinese medicine group(CM).N group received no special treatment;WM group received Compound lactic acid bacteria 0.66g tid,and CM group received Jianpi Mixture 20ml tid,and both courses are 2 months.Record the anxiety self-assessment scale and depression self-assessment scale before and after treatment in the N,WM,and CM groups;and observe the clinical symptoms of the WM and CM groups before treatment,every two weeks after medication,and after treatment,and TCM symptoms were graded quantitatively.Subsequently,stool samples of 5 healthy people in the N group,5 patients in the WM and CM groups before and after treatment were randomly selected for bacterial flora detection.The 16S rDNA high-throughput gene sequencing method was used to analyze the changes of intestinal microbiotaResults:1.Experimental research:General situation:After modeling,the weight of IBS-D mice with spleen deficiency type was significantly reduced(P<0.05),and the Bristol score was significantly increased(P<0.05).After treatment,the weight and food intake of mice in the Jianpi Mixture group increased(P<0.05),Bristol score was significantly reduced(P<0.05).Intestinal microbiota:In the study,the sequencing depth was sufficient,and the samples all had good species richness and species diversity.Compared with the control group,the species richness and species diversity of intestinal microbiota in feces and intestinal mucosa of spleen-deficiency IBS-D mice were decreased(P<0.05);spleen-deficiency IBS-D mice showed changes in composition of and in ratio of the intestinal microbiota in feces and intestinal mucosa at level of phylum,class,order,family,genus and species.Treatment with Jianpi Mixture increased the species diversity of intestinal microbiota in spleen-deficiency IBS-D mice(P<0.05),the abundance of beneficial bacteria(P<0.05),and decreased the abundance of harmful bacteria(P<0.05)at the levels of the phylum and genus.Compared with healthy mice,the species richness and species diversity of intestinal microbiota of spleen-deficiency IBS-D mice are decreased.2.Clinical research:Evaluation of clinical efficacy:The average total symptom score of CM-A was 13±3.89,and the average total symptom score of CM-B was 3.4±2.87.After the intervention of Jianpi Mixture,the symptom score decreased significantly(P<0.05).The total effective rate of TCM symptoms in the CM group was 47%and that in the WM group was 70%.The difference between the two was not statistically significant(P>0.05).From the perspective of individual symptoms,symptoms of stool changes,abdominal pain,abdominal distension,languid,mental fatigue,and bowel sounds symptoms were significantly improved after treatment in the CM group(P<0.05).Compared with the WM group,there was no significant difference in each single TCM symptom(P>0.05).There was no significant change in anxiety and depression index of CM group and WM group before and after treatment(P>.05).Intestinal microbiota:In the study,the sequencing depth was sufficient,and the samples all had good species richness and species diversity.Combined with Alpha diversity analysis,the intestinal microbiota diversity of patients with spleen deficiency type IBS-D had an increasing trend after Jianpi mixture intervention,but the difference was not significant(P>0.05),and there was no significant difference with the diversity after complex lactic acid bacteria intervention(P>0.05)).Based on the metastas analysis,Granulicatella abundance was down-regulated after Jianpi mixture intervention(P<0.05).Compared with WM-B,CM-B had lower Bacteroides abundance and higher Citrobacter abundance(P<0.05).At the phylum level,the F/B of CM-B and WM-B are mainly elevated.At the genus level,Prevotella is the main dominant bacterium in CM-A,and the enterotypes is mostly attributed to ETP;Bacteroides is the main dominant bacterium in WM-A,and the enterotypes is mostly attributed to ET B.After drug intervention,CM-B and WM-B were mainly reduced in Prevotella abundance and increased in Bacteroides abundance,but the main dominant bacteria remained unchanged,and the enterotypes did not show a tendency to change to other enterotypes.Conclusion:1.Jianpi Mixture has a better clinical effect on patients with IBS-D of spleen deficiency type,especially it can significantly improve the main symptoms such as deformity of the stool and abdominal pain.2.Jianpi mixture can improve the diversity of IBS-D intestinal microbiota,increase the abundance of beneficial bacteria and reduce the abundance of harmful bacteria,so as to adjust the intestinal microecology to achieve balance.3.The effects of Jianpi mixture and compound lactic acid bacteria on the intestinal flora of spleen deficiency type IBS-D patients and mice are basically the same.It is speculated that Jianpi Mixture may have a probiotic-like effect.