Characteristics Of Gut Microbiota In Patients With Common TCM Syndromes Of Gout

Objective:To explore the differences in abundance and diversity of intestinal flora of common TCM syndromes of gout at different classification levels,and to summarize the composition law of intestinal flora of common TCM syndromes of gout,in order to provide objective basis for TCM syndrome differentiation and classification of gout.Methods:A total of 67 patients with gout were included,including 30 patients with damp-heat syndrome and 37 patients with spleen deficiency and dampness syndrome,and30 healthy volunteers were collected as the control group.A total of 97 patients were collected for clinical information and biological samples,and real-time single molecule sequencing was conducted based on PacBio Sequel II platform.The sequencing results of each group were analyzed by classification operation unit statistics.Alpha,Beta analysis and significance analysis of difference between groups were used to compare the differences in intestinal flora of common TCM gout syndrome types.LEf Se analysis and Metastats analysis were used to identify biomarkers with significant differences between the two groups of gout syndrome types.Results:1.There are statistical differences in uric acid and triglyceride of gout patients in damp-heat accumulation syndrome group,spleen deficiency and dampness-blocking syndrome group and healthy control group(P<0.05);2.A total of 2395262 CCS sequences and 1028 OTUs were obtained from 97 samples.3.Alpha diversity analysis showed that the sample size of this sequencing was sufficient,and the microbe uniformity of each group was close,while the microbe richness of the healthy control group and the spleen deficiency and dampness syndrome group were different(P<0.05).3.Beta diversity analysis: In PCoA analysis,the interpretation of PC1 was 15.32%,and that of PC2 was 4.72%.In PC1,the three groups all had their own tendency to aggregate,but the aggregation was not obvious.NMDS Analysis stress<0.2(Anosim test: R=0.0774456,P<0.05).4.Significance analysis of differences between groups:(1)LEf Se analysis consensus of 11 species with significant differences between groups.Among them,Prevotellaceac was the most representative in the healthy control group,Negativicutes was the most representative in the damp-heat accumulation syndrome group,and clostridia was the most representative in the spleen deficiency and dampness-blocking syndrome group.(2)Metastats analysis: there were two types of bacteria with significant difference in phylum level,the most representative of which was Firmicutes;There were four classes at class level,among which the abundance of Clostridia was significantly different.There were 9 different species at the order level,among which unclassified Clostridia had the most difference.There were 24 species at the family level,and Acinetobacter sp CAG 196 had the largest abundance difference.There are 65 species classified at the generic level.unclassified Acinetobacter sp CAG 196 is the most classified species.There were 150 distinct species at the species level,of which Peptococcus sp canine taxon 334 was the most representative.Conclusions:1.There were differences in the species richness of flora in phylum,class,order,family,genus and species between gout syndrome of damp-heat accumulation and syndrome of spleen deficiency and dampness obstruction and healthy people;2.Biomarkers with significant differences existed in all the three groups.Prevotellaceae was the most representative in the healthy control group,Negativicutes was the damp-heat accumulation syndrome group,and clostridia was the spleen deficiency and dampness syndrome.3.The intestinal flora of the syndrome of dampness-heat accumulation and the syndrome of spleen deficiency and dampness-blocking have specific flora at the order,family,genus and species level,which may be one of the basis for TCM syndrome differentiation and classification of GA.