Experimental And Clinical Study On The Changes Of Intestinal Microbiota Of Spleen Deficiency IBS-D By High-throughput Sequencing

Objective:To investigate the changes of intestinal microbiota of spleen deficiency diarrhea-type irritable bowel syndrome(IBS-D)mice and patients through 16S rDNA sequencing analysis,which can be used as one of the therapeutic targets and provide the basis for the next intervention research.Methods:1.Experimental study:Totally 48 clean-grade young ICR mice were randomly divided into control check group(CK)and experimental group(IBS-D),and the models of spleen deficiency IBS-D mice were established.Subsequently,a general comparison was performed,and the intestinal microbiota structure of the subjects(15 spleen deficiency IBS-D mice and 5 healthy mice)were analyzed by 16S rDNA sequencing.2.Clinical research:20 healthy volunteers were included as control check group(CK),and 23 spleen deficiency IBS-D patients were included as experimental group(IBS-D).The typical clinical symptoms of experimental group were statistically analyzed,and the sex、age、diet、living habits、anxiety、depression of the two groups were compared.Finally,the intestinal microbiota structure of 10 spleen deficiency IBS-D patients and 5 healthy people were analyzed by 16S rDNA sequencing.Results:1.Experimental study:Compared with healthy mice,the species richness and diversity of intestinal microbiota in spleen deficiency IBS-D mice were decreased.At the six classification levels of phylum、class、order、family、genus and species,the structure of intestinal microbiota in two groups of mice were different.At the phylum level,Bacteroidetes and Firmicutes of the healthy mice were accounted for 94.14%,while Bacteroidetes and Firmicutes of the spleen deficiency IBS-D mice were accounted for 79.63%.Compared with healthy mice,the spleen deficiency IBS-D mice were characterized by decreased Bacteroidetes、Saccharibacteria、Cyanobacteria,and increased Proteobacteria and Verrucomicrobia(P<0.05).At the genus level,the healthy mice were dominated by Bacteroides、Alistipes and Lactobacillus,while the spleen deficiency IBS-D mice were dominated by Bacteroides、Akkermansia and Parasutterella.Compared with healthy mice,Alistipes、Lactobacillus、Rikenella、Candidatus-Saccharimonas、Candidatus-Arthromitus of spleen deficiency IBS-D mice were redcued,Bacteroides、Akkermansia、Parasutterella、Lachnoclostridium、Anaerotruncus were increased(P<0.05).2.Clinical research:At the six classification levels of phylum、class、order、family、genus and species,the structure of intestinal microbiota in spleen deficiency IBS-D patients and healthy people were different.At the class level,the healthy people were dominated by Bacteroidia and Clostridia,while the spleen deficiency IBS-D patients were dominated by Bacteroidia and Clostridia.Compared with healthy peolpe,the spleen deficiency IBS-D patients were characterized by increased Gammaproteobacteria(P<0.05).At the genus level,the healthy people were dominated by Bacteroides、Faecalibacterium and Blautia,while the spleen deficiency IBS-D patients were dominated by Prevotella、Bacteroides and Faecalibacterium.Compared with healthy people,Prevotella and Actinomyces of spleen deficiency IBS-D patients were increased(P<0.05).This study found two types of Enterotypes:Enterotypes 1(Enterotypes Bacteroides,ET B)and Enterotypes 2(Enterotypes Prevotella,ET P).ET B is common in healthy people,while ET P is common in spleen deficiency IBS-D patients.Conclusion:1.The intestinal microbiota of the spleen deficiency IBS-D mice and patients has changed significantly at the six classification levels,which is manifested by a decline in a large number of beneficial bacteria and an increase in a large number of harmful bacteria.2.This clinical research found two types of Enterotypes:Enterotypes 1(ET B)and Enterotypes 2(ET P).ET B is common in healthy people,while ET P is common in spleen deficiency IBS-D patients.