| Objective:Through studying the biodiversity changes of the gut microbial community in patients with T2 DM clinically treated with GLP-1 RAs,the differential flora of gut flora after using GLP-1 RAs are explored,to analyze the correlation between the main flora in the gut and glycemic control and weight control,thus provide a theoretical basis for the subsequent study of the mechanism of gut microbes in the treatment of T2 DM with GLP-1 RAs.Methods:(1)This study used its own before-and-after controlled study.Forty-one patients with newly diagnosed T2 DM were recruited as subjects.Stool,blood,and BMI were collected from the subjects before GLP-1 RAs were used,one week after GLP-1 RAs were used,and 48 weeks after GLP-1 RAs were used.Samples before the use of GLP-1 RAs were labeled as Group F,samples one week after the use of GLP-1 RAs were labeled as Group S,and samples 48 weeks after the use of GLP-1 RAs were labeled as Group T.(2)16s rRNA amplicon sequencing were used to detest the biodiversity of the intestinal flora of subjects in group F,S,and T,and compare the differences in the composition and structure of the intestinal flora of the three groups,among which significant differences in the intestinal flora might be searched for by bioinformatics.(3)The FISH technique was used to detect changes in the abundance of Faecalibacterium prausnitzii and Lactobacillus delbrueckii in the intestine before and1 week after the use of GLP-1 Ras.(4)The correlation between Faecalibacterium prausnitzii and Lactobacillus delbrueckii,and,blood glucose and BMI,was analyzed.Results:(1)The results of 16 S sequencing showed that each group of samples had an average of 13,674 ASVs,including 6,176 ASVs in group F,6,592 in group S and3,054 in group T.(2)The results of the Rarefaction Curve and the Shannon Index Curve showed that the sample size of this study was sufficient and the data were reliable,which means the number of ASVs would not increase much even if the sample size was increased further.(3)α-diversity analysis showed that there was no significant change in species abundance and diversity of intestinal flora between group S and group F,while the species abundance of intestinal flora in group T decreased significantly compared to group F,although there was no significant change in species diversity in group T.(4)β-diversity analysis showed no significant change in species composition of the intestinal flora in groups F and S,but the composition and structure of the intestinal flora in group T were significantly different from groups F and S.(5)Anosim analysis showed that the Inter-group differences in this study were significantly greater than the Intra-group differences,and the results of β-diversity analysis were reliable,with significant differences in the composition of intestinal flora between group T and groups S and F.(6)The main bacteria of the three groups F,S,and T at the phylum level were Bacteroidota,Firmicutes,and Proteobacteria;at the phylum level were Bacteroidia,Clostridia,and Gammaproteobacteria;at the order level were Bacteroidales,Lachnospirales,Oscillospirales;at the family level were Bacteroidaceae,Lachnospiraceae,Ruminococcaceae;at the genus level were Bacteroides,Blautia,Faecalibacterium;the dominant species were Bacteroides＿plebeius and Bacteroides＿vulgatus.(7)The Lef Se analysis did not find any gut microorganisms with significant differences between the three groups.(8)FISH method detected that a significant increase in the abundance of Faecalibacterium prausnitzii in the intestine after 1 week of using GLP-1 RAs,and no significant change in Lactobacillus delbrueckii.(9)A significant decrease in fasting blood glucose levels and a significant decrease in body weight in subjects after 1 week of using GLP-1 RAs.(10)Spearman’s correlation analysis showed a significant negative correlation between the abundance of Faecalibacterium prausnitzii in the intestine and fasting blood glucose after 1 week of treatment with GLP-1 RAs for T2 DM.Conclusions:(1)The composition and structure of the intestinal flora of patients showed No significant change after 1 week of clinical treatment of T2 DM with GLP-1 RAs.(2)The α-diversity analysis and β-diversity analysis of the intestinal flora of patients with T2 DM were significantly different after 48 weeks of using GLP-1 RAs compared with before and 1 week after use,with the abundance of microbial communities in the intestine was significantly reduced,while the diversity was not changed.(3)Lef Se analysis did not find gut microorganisms with significant differences in the gut before the use of GLP-1 RAs,1 week after the use of GLP-1 RAs,and 48 weeks after the use of GLP-1 RAs.(4)FISH was used to detect changes in intestinal flora after 1 week of GLP-1RAs use and found a significant increase in the abundance of Faecalibacterium prausnitzii and a negative correlation with fasting glucose in T2 DM patients. |
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