Application of intestinal bacteria in ulcerative colitis prediction technology

Abstract

The invention discloses an application of intestinal bacteria in an ulcerative colitis prediction technology, and belongs to the technical field of biology, and the intestinal bacteria are Alistipes finegoldii; the method comprises the following steps: detecting the relative abundance of Alistipes finegii in an excrement sample of a patient, and using the low relative abundance of Alistipes finegii to predict the disease activity of ulcerative colitis; according to the application disclosed by the invention, not only is the association between the Alistipes finegili and the UC disease activity verified, but also the potential action mechanism of the Alistipes finegili is further explored. By means of 16s amplicon sequencing and transcriptomics combined analysis, it is found that the Alistipes finegili plays an anti-inflammatory role through an Hsp90 / AMPK / ACC signal channel, and the intestinal barrier function is maintained. The invention verifies that a new view angle is provided for the effect of the Alistipes finegili in the UC pathogenesis, and provides a theoretical basis for developing a treatment strategy based on the strain.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, in particular to the application of intestinal bacteria in the prediction technology of ulcerative colitis. Background Art

[0002] Inflammatory bowel disease (IBD) is an idiopathic intestinal inflammatory disease that affects the ileum, colon, and rectum. It includes two subtypes: ulcerative colitis (UC) and Crohn's disease (CD). Compared with CD, UC is more common and can occur in people of all ages.

[0003] The pathogenesis of UC is still unclear and the therapeutic effect is poor. As the disease course prolongs, drug resistance and canceration rates increase. No effective early warning molecules for the disease have been found, so it is urgent to explore the pathogenesis of UC and explore new targets.

[0004] Microbiome research is developing rapidly. Intestinal flora plays an important role in the occurrence and development of UC. Studies have shown that intestinal flora imbalance in patients with active UC is manifested as an increase in potential pro-inflammatory and harmful pathogens, which interact with intestinal mucosal immunity through their metabolites, jointly inducing the occurrence of intestinal inflammatory microenvironment, thereby causing intestinal barrier function damage. Among them, lipopolysaccharides produced by intestinal Gram-negative bacteria and harmful molecules metabolized by other pathogenic microorganisms can also enter the circulatory system through the damaged intestinal barrier, aggravating local inflammation under repeated stimulation, and then involving other systems. For this reason, the application of intestinal bacteria in the prediction technology of ulcerative colitis is proposed.

[0005] The above information disclosed in this background technology is only used to increase the understanding of the background technology of the present invention and therefore, it may include information that does not constitute the prior art known to ordinary technicians in this field. Summary of the invention

[0006] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, one object of the present invention is to propose the application of intestinal bacteria in the prediction technology of ulcerative colitis.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] Application of intestinal bacteria in ulcerative colitis prediction technology: the intestinal bacteria is Alistipes finegoldii;

[0009] Among them, the relative abundance of Alistipes finegoldii in patient stool samples was detected, and the low relative abundance of Alistipes finegoldii was used to predict the disease activity of ulcerative colitis;

[0010] The predicted mechanism involves Alistipes finegoldii acting through the Hsp90 / AMPK / ACC signaling pathway as a key early warning molecule for ulcerative colitis disease activity.

[0011] As a further optimization scheme of the present invention, 16S amplicon sequencing and transcriptomics combined analysis were used to quantify the relative abundance of Alistipes finegoldii in fecal samples;

[0012] The combined analysis results were used to evaluate the status of the Hsp90 / AMPK / ACC signaling pathway to predict the disease activity of ulcerative colitis.

[0013] Compared with the prior art, the present invention has the following beneficial effects:

[0014] The present invention confirms through clinical sample analysis and animal experiments that the low relative abundance of Alistipes finegoldii can effectively predict the disease activity of UC, providing more accurate targets and predictive indicators for FMT treatment.

[0015] This study not only verified the association between Alistipes finegoldii and UC disease activity, but also further explored its potential mechanism of action. Through 16S amplicon sequencing and transcriptomics combined analysis, it was found that Alistipes finegoldii exerted anti-inflammatory effects through the Hsp90 / AMPK / ACC signaling pathway and maintained intestinal barrier function. The verification of the role of Alistipes finegoldii in the pathogenesis of UC provides a new perspective and provides a theoretical basis for the development of therapeutic strategies based on this species.

[0016] This paper uses a combination of 16S amplicon sequencing, transcriptomics, ROC model and other omics technologies and bioinformatics analysis methods to deeply explore the changes in the intestinal flora of UC and the mechanism of action of Alistipes finegoldii from multiple dimensions. The integrated application of multiple omics technologies improves the reliability and comprehensiveness of the research and provides more abundant information for the diagnosis and treatment of UC.

[0017] The above summary is for illustrative purposes only and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments and features described above, further aspects, embodiments and features of the present invention will be readily apparent by reference to the accompanying drawings and the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a diagram showing the difference in intestinal flora between healthy volunteers and UC patients in the present invention;

[0019] Figure 2 This is a diagram showing the difference in intestinal flora between active UC patients (UC-Ac) and remission UC patients (UC-Re) in the present invention;

[0020] Figure 3 A schematic diagram of Alistipes finegoldii as an independent predicted molecule of UC in the present invention;

[0021] Figure 4 Schematic diagram of intestinal flora of mice with enteritis in remission after treatment (DSS-Ac) and mice with enteritis without treatment (DSS-Re);

[0022] Figure 5 Schematic diagram of 16s amplicon sequencing and transcriptome analysis of DSS-Ac and DSS-Re in the present invention;

[0023] Figure 6 This is a diagram showing the effect of Alistipes finegoldii in experimental enteritis in the present invention;

[0024] Figure 7 This is a diagram showing the effect of Alistipes finegoldii in the present invention in inhibiting enteritis through the Hsp90 / AMPK / ACC pathway. DETAILED DESCRIPTION

[0025] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Embodiment 1

[0027] As attached Figure 1 As shown, in area A, the Shannon index (one of the best evaluation indicators of α-diversity uniformity and richness) in the intestinal flora of UC patients was significantly lower than that of the healthy control group;

[0028] Area B: PCoA analysis suggested that there were differences in the diversity of intestinal flora between UC patients and healthy volunteers (β-diversity);

[0029] Area C-E: The combined use of Welch's T test, Indicator analysis and LDA analysis showed that Alistipes was significantly reduced in UC patients and significantly increased in healthy volunteers.

[0030] Embodiment 2

[0031] As attached Figure 2 As shown, area A: the Shannon index (one of the best evaluation indicators of α-diversity uniformity and richness) in the intestinal flora of UC-Ac patients was significantly lower than that of the healthy control group;

[0032] Area B: PCoA analysis showed that there were differences in the diversity of intestinal flora between UC-Ac patients and UC-Re patients (β-diversity);

[0033] Area C-D: The combined use of Indicator analysis and LDA analysis showed that Alistipes was significantly reduced in UC-Ac patients and significantly increased in UC-Re patients.

[0034] Embodiment 3

[0035] As attached Figure 3 As shown in the figure, the ROC model was used to analyze the clinical indicators such as the differential bacterial genera / species and Mayo score of UC patients in the active stage vs the remission stage, and the AUC values, specificity and sensitivity of 15 differential bacterial genera and 3 differential bacterial species were calculated. It was found that Alistipes finegoldii can be used as an independent predictor of UC activity.

[0036] Embodiment 4

[0037] As attached Figure 4 As shown, in order to further clarify the different bacterial species, a DSS-induced enteritis mouse experiment was conducted, mainly to compare the differences in intestinal flora between enteritis mice in the disease remission period after treatment and untreated enteritis mice.

[0038] Area A: The OTUs in the healthy control group (Control) were more than those in the two groups of enteritis mice, and DSS-Re was more than DSS-Ac;

[0039] Area B: The results of the dominant species heat map at the species level indicated that Alistipes finegoldii decreased significantly during the active period of enteritis;

[0040] Zone C-D: Indicator analysis at the genus and species levels suggested that the number of Alistipesfinegoldii in the genus Alistipes was significantly reduced during the active stage of enteritis.

[0041] Embodiment 5

[0042] As attached Figure 5As shown, 16S amplicon sequencing and transcriptome analysis of intestinal tissues were performed on the intestinal contents of the DSS-Ac vs. DSS-Re groups of mice, and the target signaling pathways were screened out based on the differential KEGG enrichment in the reference RNA seq: AMPK signaling pathway (ko04152), Estrogen signaling pathway (ko04915); the target genes were screened out based on the differential test FDR value: Hspa1b, Hspa1a, Hsp90ab1, Gpd1, Fasn, Eif2s2, Cytip, COL5A3, Clec7a, Ccdc122, Casp4, Bmp5, Apobec1, Apoa1, Acaca, etc.

[0043] KEGG analysis showed that the AMPK (Scd2, Fasn, Acaca, irs1, Creb3l1) signaling pathway was enriched (A). The association analysis of the two groups found that Alistipes was significantly correlated with the expression of key genes in the AMPK signaling pathway (B).

[0044] Embodiment 6

[0045] As attached Figure 6 As shown, area A: Alistipes finegoldii significantly reduced the disease activity (DAI) of enteritis mice;

[0046] Area B-C: Alistipes finegoldii significantly improved the degree of colon shortening in enteritis mice;

[0047] Region D: Alistipes finegoldii significantly downregulated the expression of the classic inflammatory pathway NFκB;

[0048] Area E: Alistipes finegoldii significantly improved colon pathological damage in enteritis mice.

[0049] Embodiment 7

[0050] As attached Figure 7 As shown, area A: Alistipes finegoldii was found to significantly activate the Hsp90 / AMPK / ACC pathway at the mRNA level;

[0051] Area B-C: Immunohistochemistry of mouse colon tissue indicated that Alistipes finegoldii significantly increased the protein content of Hsp90 / AMPK / ACC.

[0052] Parts not involved in the present invention are the same as the prior art or can be implemented by the prior art. Although the embodiments of the present invention have been shown and described, it is understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Application of intestinal bacteria in ulcerative colitis prediction technology, characterized by: The intestinal bacteria is Alistipes finegoldii; Among them, the relative abundance of Alistipes finegoldii in patient stool samples was detected, and the low relative abundance of Alistipes finegoldii was used to predict the disease activity of ulcerative colitis; The predicted mechanism involves Alistipes finegoldii acting through the Hsp90 / AMPK / ACC signaling pathway as a key early warning molecule for ulcerative colitis disease activity.

2. The use of intestinal bacteria in the prediction technology of ulcerative colitis according to claim 1, characterized in that: Combined 16S amplicon sequencing and transcriptomic analysis was used to quantify the relative abundance of Alistipes finegoldii in fecal samples; The combined analysis results were used to evaluate the status of the Hsp90 / AMPK / ACC signaling pathway and predict the disease activity of ulcerative colitis.

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