Characteristics Of Gut Microbiome In Patients With Common Traditional Chinese Medicine Syndromes Of Osteoarthritis

Objective:To explore the differences of intestinal flora in different TCM syndromes of Osteoarthritis(OA),analyze the sources of the differences,and summarize the composition rules of intestinal flora in different TCM syndromes of OA,in order to deepen the understanding of the essence of OA TCM syndromes and provide an objective research basis for the classification of OA TCM syndromes.Method:A total of 75 OA patients were enrolled,including 15 cases of qi and blood deficiency syndrome,damp-heat blocking syndrome,phlegm and blood stasis syndrome,cold-damp obstruction syndrome,liver and kidney deficiency syndrome,and 15 healthy volunteers were collected as control group.A total of 90 patients were enrolled for clinical information and biological samples collection,and single-molecule real-time sequencing based on Pac Bio Sequel II platform.The sequencing results of each group were analyzed by classification operation unit statistics,Alpha and Beta diversity analysis,and significant difference analysis between groups to compare the differences of intestinal flora in different TCM syndromes of OA.Lef Se analysis was used to identify the biomarkers with significant differences in each OA syndrome group.Results:1.The age of the OA patients included in this study ranged from 53 to 69 years,and the male to female ratio was 1:3.2.There was no significant difference in age,gender and BMI between the OA syndrome groups and the healthy control group(P>0.05).2.A total of 1,289,415 valid sequences were obtained from 90 samples,generating 3650 OTUs.3.Alpha diversity analysis showed that the coverage of sample species was reasonable and the amount of sequencing was sufficient,and the microbial evenness of each group was similar.Microbial richness was significantly higher in healthy controls than in OA groups.4.Beta diversity analysis:in PCA analysis,the interpretation of PC1 was 25.50%,and the interpretation of PC2 was 17.73%.On PC2,the distribution distance between OA syndromes and healthy controls was far,while the distribution within the group was close.In PCo A analysis,the interpretation of PC1 was 25.40%.Each OA syndrome group and the healthy control group had their own aggregation tendency,but the aggregation was not obvious.NMDS analysis Stress<0.2.Statistical test:PERMANOVA test:R~2=0.083,P=0.001;Anosim test:R=0.091,P=0.001.5.At the phylum level,four distinct taxa were identified,with the most significant one being Verrucomicrobia.There were five classes at the class level,of which Bacilli had the largest difference in abundance.There were 7kinds of significant difference species at the order level,with Lactobacillales being the most representative.At the family level,there were 15 types of bacteria with significant differences in abundance,among which Selenomonadaceae had the largest difference.At the genus level,there were 33 species with significant differences in abundance,among which Faecalibacterium was the most different.A total of 58 species with significant differences in abundance were obtained at the species level,with Akkermansia muciniphila having the largest difference.6.LEf Se analysis identified 26 significant species.The most representative biomarker of qi and blood deficiency syndrome group was Parasutterella in Proteobacteria.There were a total of 18 biomarkers in the damp-heat blocking collaterals syndrome group,and Streptococcus was the most representative one.There were three biomarkers in the syndrome group of phlegm and blood stasis,and Bacteroides＿vulgatus was the most representative one.The cold-damp obstruction syndrome group had 6biomarker,and the most representative biomarker was Bifidobacterium adolescentis.There were 10 biomarkers in the liver-kidney deficiency syndrome group,the most representative of which was Campylobacterales.Conclusion:1.Compared with the healthy control group,the OA patients with different TCM syndromes had a significant decrease in the richness of gut microbiota,and there were significant differences in the composition of gut microbiota among the OA patients with different TCM syndromes.2.The distribution structure of high-abundance flora in each group was similar at phylum,class,order,family,genus,and species levels,but there were significant differences in low-abundance flora at each taxonomic level.3.There were significant differences in biomarkers among all groups.The group of qi and blood deficiency syndrome was Parasutterella;The damp-heat blocking collateral syndrome group was Streptococcus of Firmicutes.Bacteroides＿vulgatus syndrome group;The cold-dampness obstruction syndrome group was Bifidobacterium＿adolescentis;In the liver-kidney deficiency syndrome group was Campylobacterales.