Multiomics Study On Intestinal Flora And Metabolism Of Patients With Ulcerative Colitis In Xinjiang

Objective: To find the difference between intestinal flora and fecal metabolites in Uygur and Han patients with ulcerative colitis(UC),to screen the biomarkers of fecal metabolism level and the bacteria with disease identification ability in UC patients,and to analyze the internal relationship between intestinal flora and metabolism difference between Uygur and Han UC patients.The role of different bacteria and metabolites in UC inflammation was studied by animal model construction.Methods: 1)There were 23 newly diagnosed Uygur patients with ulcerative colitis and 25 Han patients with ulcerative colitis.Healthy spouses or first-degree relatives who have lived together for more than 1year with the patient group are selected to match the patients 1: 1 as a control group.The provided stool samples were amplified by the highly variable region of bacterial16 Sr DNA,the Miseq library was constructed and IIIumina was sequenced,and the RDP classifier was used to classify and annotate each sequence.Then,the classification of samples was obtained by comparing with sliva database.Analyzed the microbial diversity and abundance index among the groups,and screened out bacteria with significant differences in abundance among the groups.Completed taxonomy and species structure analysis,and further screened bacteria with disease identification ability through random forest analysis.All metabolites were extracted from the same stool sample as in the first stage.The LC-MS metabolomics analysis platform was used to quantitatively identify the differential metabolites.Objective to search for different metabolites among groups and screen biomarkers of metabolic level in UC feces.Based on the results of microbial diversity and metabonomics experiments,the correlation between differentially expressed microorganisms and metabolites in Uygur and Han patients with UC was analyzed by bioinformatics analysis.3)The UC rat model was established,and the fecal bacteria transplantation of Uygur and Han ulcerative colitis fecal bacteria liquid was carried out to UC model rats and normal rats,respectively.The colonic mucosal tissues were collected to detect the expression of UC-related cytokines(TNF-α,IL-4,IL-6,IL-10).Results: 1)The results of microbial diversity analysis showed that there were significant differences in microbial community diversity between Uygur UC patients and normal Uygur group,Uygur normal group and Han normal group,Han UC patients and Uygur normal group.The microbial diversity of Uygur UC patients was significantly lower than that of the normal control group(P < 0.05).However,there was no significant difference in microbial diversity and richness index between Han UC patients and their relatives(P>0.05).The abundance of Burkholderia-Caballeronia-Paraburkholderia in the Uygur UC group was significantly higher than that in the Han UC group,and the abundance of Faecalibacterium,Bifidobacterium and Blautia in the Han UC group were significantly higher than that of the Uygur UC group(P< 0.05).Compared with the Uygur normal control group,the abundance of Veillonella was significantly higher in the Uygur UC group,while the abundance of Subdoligranulum and Ruminococcaceae＿UCG-002 was significantly lower(P<0.05).The abundance of Prevotella＿9 in the Han UC group was significantly higher than that in the Han normal control group.While Blautia,Anaerostipes and [Eubacterium]＿hallii＿group were significantly lower than the Han normal control group(P<0.05).Through ROC analysis,six important microbial species were screened to have certain ability to discriminate Uygur UC disease status and healthy controls: Christensenellaceae＿R＿7＿group,Ruminococcaceae＿UCG＿005,Ruminococcaceae＿UCG＿010,Ruminococcaceae＿UCG＿013,Haemophilus and Ezakiella.2)Partial least squares discriminant analysis(PLS-DA)found that there were significant differences in metabolites between groups.Two potential biomarkers that can be annotated were screened between the Han UC group and Han normal control group.In the Uygur UC group 21 potential biomarkers that can be annotated were screened out between the Uygur normal control group.Omega-3 arachidonic acid as a potential biomarker was significantly reduced in Uygur UC patients.In the association analysis of microbial diversity and metabonomics,Haemophilus was found to be highly negatively correlated with N-acetylgalactosamine in Uygur patients with UC.3)Compared with the control group,the levels of TNF-α and IL-6 in the two groups were significantly increased(P<0.05),and IL-4,IL-10 were significantly increased after the fecal bacteria liquid of Uygur and Han UC patients was treated with ulcerative colitis model rats(P<0.05).There was no significant difference in the levels of TNF-α,IL-10 and IL-4 between the Uygur and Han fecal bacterial transplantation groups(P>0.05),but the IL-6 level of the Uygur UC group was significantly higher than that of the Han group(P < 0.05).No significant changes in the expression levels of inflammatory factors were observed in the enema treatment group with fecal bacteria liquid of the two ethnics of normal rats.Conclusion:There are differences in the structure and diversity of gut microbes between Uygur and Han UC patients.Christensenellaceae＿R＿7＿group,Ruminococcaceae＿UCG＿005,Ruminococcaceae＿UCG＿010,Ruminococcaceae＿UCG＿013,Haemophilus and Ezakiella6 microbial species have certain diagnostic ability for Uygur UC patients and healthy controls.There are significant differences in intestinal fecal metabolites between Uygur UC patients and Han UC patients.Two potential biomarkers that can be annotated were screened between the Han UC group and the normal control group.The Uygur UC group and the normal control group 21 potential biomarkers that can be annotated were screened among the groups.Omega-3 arachidonic acid as a potential biomarker was significantly reduced in Uygur UC patients.Haemophilus is likely to affect the intestinal mucosal barrier by affecting the expression of N-acetylgalactosamine.The influence of differences in gut microbes and metabolomics on UC disease may be related to the expression level of IL-6.The difference in intestinal microbial structure,the interaction between flora disorders and metabolic levels,and the role of Omega-3,IL-6,and NF-κb signaling pathways may be related to the special clinical manifestations and genetic background of Uyghur UC patients in Xinjiang.The level of intestinal microbes and fecal metabolites may be the promoting factors of ulcerative colitis disease rather than the initiating factors.