Study On The Characteristics Of Intestinal Flora In Patients With Chronic Spontaneous Urticaria And Gastrointestinal Damp-heat Syndrom

Objective To investigate the characteristics of gut microbiota in patients with chronic spontaneous urticaria(CSU)and gastrointestinal damp-heat syndrome in Beijing,16S rRNA high-throughput sequencing technology was used to compare and analyze the species composition of gut microbiota between the CSU patients and healthy individuals.The results aim to provide supplementary methods and theoretical basis for the treatment and prevention of this disease.Methods Based on the inclusion and exclusion criteria,15 patients with CSU and gastro-intestinal dampness-heat syndrome(CSU group)who were treated or recruited at the dermatology clinic of Dongfang Hospital,Beijing University of Chinese Medicine from April 2022 to March 2023,and 10 healthy volunteers(HC group)were selected.Fresh fecal samples were collected from the subjects,and DNA extraction,PCR amplification,library construction,ASV denoising,species annotation and other steps were performed.Bioinformatics analysis including alpha diversity analysis,beta diversity analysis,analysis of species composition and differences between the two groups,and LEfSe analysis were conducted.Results1.Baseline data:The gender,age,and BMI of the subjects were evenly distributed between the two groups,and there were no statistically significant differences observed(P>0.05).2.Following ASV denoising,a total of 8684 ASVs were obtained,consisting of 7360 ASVs in the CSU group,881 ASVs in the HC group,and 443 ASVs shared between both groups.3.Alpha diversity analysis showed that,among the various statistical analysis indices,the Chao1 index and observed index of the CSU group significantly increased(P<0.01)compared to the control group,while the pielou e-index significantly decreased(P<0.01),indicating a statistically significant difference.There was no significant difference observed between the two groups in the dominance index and goods coverage index,as well as the Shannon and Simpson indices(P>0.05).4.Beta diversity analysis revealed that,in the UPGMA clustering tree,samples from the same group tended to cluster together,while samples from different groups were more likely to be dispersed into different branches.Additionally,the PCoA visualization demonstrated that the samples from both groups were clustered together,indicating similar origins.5.The analysis of differences in species composition revealed that,at the phylum level,the abundance of Bacteroidota was significantly decreased(P<0.05)in the CSU group compared to the healthy control group,while the abundance of Proteobacteria,Actinobacteriota,Acidobacteriota,Verrucomicrobiota,Deferribacterota,Chloroflexi,Myxococcota,and Gemmatimonadota was significantly increased(P<0.05).At the class level,the abundance of Bacteroidia was significantly decreased(P<0.05)in the CSU group,while the abundance of Bacilli,Actinobacteria,Alphaproteobacteria,Coriobacteriia,Acidobacteriae,Verrucomicrobiae,Deferribacteres,and Polyangia was significantly increased(P<0.05).Similarly,at the order level,the abundance of Bacteroidales was significantly decreased(P<0.05)in the CSU group,while the abundance of Bifidobacteriales,Lactobacillales,Erysipelotrichales,Coriobacteriales,Burkholderiales,Pasteurellales,Acetobacterales,and Vibrionales was significantly increased(P<0.05).At the family level,the abundance of Bacteroidaceae,Veillonellaceae,and Pasteurellaceae was significantly decreased in the CSU group(P<0.05),while the abundance of Bifidobacteriaceae,Enterococcaceae,Erysipelotrichaceae,Streptococcaceae,and Acetobacteraceae was significantly increased(P<0.05).Similarly,at the genus level,the abundance of Bacteroides,Faecalibacterium,and Dialister was significantly decreased in the CSU group(P<0.05),while the abundance of Bifidobacterium,Blautia,Enterococcus,Streptococcus,Turicibacter,and Eubacterium_hallii was significantly increased(P<0.05).Finally,at the species level,the abundance of Bacteroides_coprocol,Bacteroides_eggerthii,Dialister_succinatiphilus,Bacteroides_dorei,Prevotellaceae_bacterium,Alistipes_shahii,and Haemophilus_parainfluenzae was significantly decreased in the CSU group compared to the healthy control group(P<0.05),while the abundance of Blautia_faecis and Streptococcus_salivarius was significantly increased(P<0.05)6.LEfSe analysis identified 24 potential biomarkers,with the highest to lowest impact biomarkers in the CSU group being Bacilli,Actinobacteriota,Blautia,Actinobacteria,Bacteroides,Lactobacillales,Alphaproteobacteria,Enterococcaceae,Enterococcus,and Erysipelotrichales.Conversely,the most influential biomarkers in the HC group,in descending order,were Bacteroidales,Bacteroidia,Bacteroidota,Bacteroides,Bacteroidaceae,Oscillospirales,Faecalibacterium,Prevotellaceae,Prevotella copri,Bacteroides coprocola,Negativicutes,Veillonellales-selenmonadales,Bacteroides plebeius,and Veillonellaceae.Conclution1.In comparison to the healthy population,CSU patients with gastrointestinal damp heat syndrome have exhibited an increase in intestinal flora species diversity and a decrease in evenness.2.The species composition and structure of gut microbiota in CSU patients with gastrointestinal dampness heat syndrome have undergone significant alterations in comparison to the healthy population.3.A total of 24 potential biomarkers for CSU patients with gastrointestinal dampness heat syndrome were identified through LEfSe analysis.