| ObjectivesTo analyze the the difference of gut microbiota structure in patients with chronic kidney disease stage 4～5 group and control group by high throughput 16 S rDNA sequencing.To explore the possible role of gut microbiota in the development of chronic kidney disease,so as to provide a new target for the treatment of chronic kidney disease.Methods1.A total of 39 patients with stage4～5(CKD 4～5)in our hospital,including19 males and 20 females,were collected.A total of 40 healthy subjects from the physical examination center of the hospital during the same period were collected as the control group,including 18 males and 22 females.All subjects gave informed consent.Clinical data,stool samples and blood samples of each subject were collected,and trimethylamine oxide(TMAO),an important metabolite of microflora,was detected in blood by enzyme linked immunosorbent assay(ELISA).SPSS 23.0was used for statistical analysis.2.To analyze the diversity of gut microbiota in patients with chronic kidney disease stage 4～5 group and control group,high-throughput 16 S ribosomal DNA(16S rDNA)detection was used to analyze the V3-V4 conserved regions through the Mi Seq PE300 sequencing platform to understand the structure of fecal microbiota.Software such as QIIME and R language Stats package were used for statistical analysis of the differences of bacterial communities.Results1.Univariate analysis of general situation of gender and age between CKD4～5 group and healthy control group showed no significant difference(P > 0.05),showing comparability.At the same time,the level of Trimethylamine n-oxide(TMAO)between the two groups was higher in CKD4～5 group,and the difference was significant(P < 0.05).2.In the CKD 4 ～ 5 group and the control group,a total of 795 operational taxonomic units(OTUs)shared between groups or specific to each group were obtained,among which 255 OTUs with significant differences between the two groups were identified(p<0.05).The number of Final OUS in CKD 4～5 groups were lower than those in the control group,indicating that the control group had more abundant colony abundance than CKD 4 ～5 groups.The indexes of Chao1,Observe,PD whole tree,Shannon and Simpson of fecal colonies in the CKD 4～5group were relatively lower than those in the control group(p<0.05),suggesting that the diversity of species in the control group was greater.Adonis differential analysis(the number of samples in each group ≥5)showed that there were significant differences in the microbial community structure between the two groups(p<0.05),which suggested that the diversity of gut microbiota was highly correlated with CKD4 ～ 5.Additionally,61 genera with differences in the two groups were identified(p<0.05),and 111 species with significant differences in phylum,class,order,family and genus between the two groups were identified(p<0.05).The differential bacterial genera of a greater contribution in the control group were displayed in descending order: c＿Bacteroidia,o＿Bacteroidales,p＿Bacteroidetes,c＿Clostridia,o＿Clostridiales,f＿Ruminococcaceae,f＿Prevotellaceae,g＿Prevotella,g＿Faecalibacterium and g＿Roseburia,etc.While those in the CKD 4-5 group were displayed in following descending order: p＿Proteobacteria,f＿Enterobacteriaceae,o＿Enterobacteriales,c＿Gammaproteobacteria,c＿Bacilli,o＿Lactobacillales,g＿Escherichia＿Shigella,g＿Enterococcus,f＿Enterococcaceae and f＿Lactobacillaceae,etc.ConclusionsThere are abundant symbiotic communities in the human intestine.CKD4 ～ 5patients have rich bacterial diversity and different bacterial community structure.There is significant difference in intestinal microbial community composition between CKD4～5 group and healthy control group.Proteobacteria,Enterobacteriaceae,Enterobacteriales and other bacteria genera and their metabolites may be involved in the pathogenesis of CKD4 ～ 5.It is suggested that regulating intestinal flora as an intervention means may play an important role in improving the prognosis and improving the quality of life of patients with CKD. |
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