Ackermania muciniphila YG2645 with function of regulating intestinal-immune-metabolic axis and application of Ackermania muciniphila YG2645

Abstract

The invention relates to the field of microorganisms, in particular to Akkermansia muciniphila YG2645 with an intestinal tract-immune-metabolic axis regulating function, an inactivated thallus, an extract, a culture, a culture supernatant, a fermentation product and / or a fermentation supernatant of the Akkermansia muciniphila YG2645, a composition containing the Akkermansia muciniphila YG2645 and a preparation method of the composition containing the Akkermansia muciniphila YG2645 and the composition containing the Akkermansia muciniphila YG2645 and the composition containing the composition. The invention also relates to application of the compound in preparation of products for preventing and / or treating inflammatory diseases. The Akkermansia muciniphila YG2645 and related products of the Akkermansia muciniphila YG2645 have the functions of regulating and controlling immune response, regulating and controlling metabolism, regulating intestinal flora and the like, and the Akkermansia muciniphila YG2645 and the related products of the Akkermansia muciniphila YG2645 have remarkable application value and market potential.

Description

Technical Field

[0001] The present invention relates to the field of microorganisms, and specifically to a strain of Akkermansia muciniphila YG2645 having the function of regulating the intestinal-immune-metabolic axis, its derivatives, a composition comprising the aforementioned, and the use of Akkermansia muciniphila or its derivatives and the composition comprising the aforementioned in the preparation of anti-inflammatory products. Background Art

[0002] The imbalance of intestinal flora is an important factor in the occurrence of various inflammatory diseases (such as inflammatory bowel disease, rheumatoid arthritis, systemic lupus erythematosus, etc.). Normal intestinal flora regulates the host immune system through various mechanisms to maintain the balance of inflammation. Th1 and Th17 cells are two important types of pro-inflammatory T cells. Th1 cells mainly secrete IFN-γ and participate in cell-mediated immune responses; Th17 cells secrete IL-17 and participate in the pathological process of inflammatory diseases. Both play a key role in many inflammatory diseases.

[0003] Traditional treatments for inflammatory diseases rely on drugs, such as nonsteroidal anti-inflammatory drugs (NSAIDs) and immunosuppressants, but these methods may cause side effects, and long-term use may cause side effects, such as gastrointestinal discomfort, immunosuppression, etc., affecting the overall health of patients. They are also unable to specifically regulate intestinal flora or metabolic pathways, let alone enhance intestinal barrier function.

[0004] Biological agents (such as monoclonal antibodies, immunosuppressants, etc.) can effectively alleviate inflammatory responses by targeting specific inflammatory mediators or cells. However, such treatments are costly, may cause serious side effects, and require medical monitoring under specific conditions.

[0005] Some probiotics (such as lactic acid bacteria, bifidobacteria, etc.) have been used to regulate intestinal flora and treat certain inflammatory diseases. However, these probiotics mainly work by competitively inhibiting harmful bacteria or enhancing host immune responses, and lack the function of specifically regulating specific immune pathways or metabolic pathways.

[0006] Akkermansia muciniphila is an intestinal probiotic that plays a key role in human intestinal health, especially in the maintenance of the mucus layer and the regulation of the immune system. Currently, there are no reports showing its role in inflammatory diseases.

[0007] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention and should not be regarded as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art. Summary of the invention

[0008] Purpose of the Invention

[0009] The object of the present invention is to provide a strain of Akkermansia muciniphila YG2645 having the function of regulating the intestinal-immune-metabolic axis, its derivatives, a composition comprising the aforementioned, and the use of Akkermansia muciniphila or its derivatives and the composition comprising the aforementioned in the preparation of anti-inflammatory products.

[0010] Solution

[0011] In order to achieve the above object, the present invention provides the following technical solutions:

[0012] In the first aspect, the present invention provides a strain of Akkermansia muciniphila YG2645 or a derivative thereof, which is classified and named Akkermansia muciniphila and is deposited in the General Microbiology Center of China Culture Collection Administration. The address of the depository is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The deposit date is May 10, 2024, and the deposit number is CGMCC No. 30598.

[0013] The Akkermansia muciniphila YG2645 of the present invention is isolated from a fecal sample of a healthy child and is identified as Akkermansia muciniphila through 16S rRNA sequencing.

[0014] In some embodiments, the 16S rRNA gene sequence of Akkermansia muciniphila YG2645 is shown as SEQ ID NO:1.

[0015] The colony characteristics of the strain include: after culturing on BHI agar medium for 24 to 48 hours, the diameter is between 0.01 mm and 1.0 mm, the colony edge is smooth, milky white, and viscous.

[0016] The inventors have discovered and experimentally confirmed that the Akkermansia muciniphila YG2645 strain (also referred to as YG2645) of the present invention has unique metabolic regulation and immunomodulatory functions, for example, it can accurately regulate the Th1 / Th17 cell balance and linoleic acid / bile acid metabolism. Specifically, the Akkermansia muciniphila YG2645 of the present invention has the following functions:

[0017] 1) Regulate immune response

[0018] On the one hand, it regulates the balance of Th1 / Th17 cells, for example, it can be manifested as specifically regulating the ratio of Th1 and Th17 cells, reducing excessive inflammatory responses, and thus alleviating the symptoms of inflammatory diseases; on the other hand, it promotes the production of anti-inflammatory factors, such as IL-10, and inhibits the release of inflammatory factors, such as IL-8 and IL-6.

[0019] 2) Regulate metabolism

[0020] Linoleic acid metabolism: Regulates the metabolic pathway of linoleic acid, reduces the production of its oxidation products, and alleviates inflammation caused by oxidative stress.

[0021] Bile acid metabolism: By regulating bile acid synthesis and conversion, the level of inflammatory secondary bile acids is reduced and the stability of the intestinal microenvironment is maintained.

[0022] 3) Regulate intestinal flora

[0023] Targeted regulation of bacterial flora combination: YG2645 can target and regulate the increase of bacterial flora combinations that are beneficial to inflammatory diseases (such as Ubosiella, Lachnospiraceae_NK4A136_group, Bifidobacterium, Lactobacillus, Limosilatobacillus, Ligilactobacillus, etc.), and inhibit harmful bacteria (such as Ileibacterium, Alistipes, Helicobacter, Faecalibaculum, Turicibacter, etc.).

[0024] Through the application of YG2645, the intestinal-immune-metabolism axis can be precisely regulated, the intestinal flora or metabolic pathways can be specifically regulated, and the intestinal barrier function can be enhanced, providing an innovative and effective biological treatment for inflammatory diseases, with significant clinical value and market potential. Moreover, this probiotic reduces the persistence of inflammation by regulating the immune response, key metabolic pathways and intestinal flora, and provides a comprehensive treatment plan. As a probiotic treatment, it has the advantages of high safety, low side effects, and relatively low cost.

[0025] In some feasible embodiments, the derivative is the inactivated bacteria, extract, culture, culture supernatant, fermentation product, fermentation supernatant or a combination thereof of Akkermansia muciniphila YG2645.

[0026] In a second aspect, the present invention provides a composition comprising an effective amount of a first active component, wherein the first active component comprises Akkermansia muciniphila YG2645 or a derivative thereof as described in the first aspect above.

[0027] In some embodiments, the composition is a pharmaceutical composition, a food composition, or a feed composition.

[0028] In some embodiments, the composition further comprises a second active ingredient.

[0029] In some embodiments, the second active component comprises a probiotic, a postbiotic, a prebiotic, an antimicrobial agent, an immunomodulator, an anticancer agent, an inflammatory therapeutic agent, or a combination thereof.

[0030] In some embodiments, the composition is a pharmaceutical composition, which further comprises a pharmaceutically acceptable carrier and / or excipient.

[0031] In some embodiments, the pharmaceutical composition is formulated as a dosage form for nasal, oral or intragastric administration, for example, in the form of pills, tablets, lozenges, lyophilized powders, granules, capsules, aqueous solutions, alcoholic solutions, oily solutions, syrups, emulsions, suspensions, suppositories, solutions for injection or infusion, ointments, gels, tinctures, creams, patches, lotions, sprays, aerosols, powder sprays, effervescent tablets, transdermal therapeutic systems, microcapsules, implants or sticks.

[0032] In a third aspect, the present invention provides the use of Akkermansia muciniphila YG2645 or its derivatives as described in the first aspect, and the composition as described in the second aspect in the preparation of a product, preferably a pharmaceutical product, for the following purposes:

[0033] 1) Regulate the balance of Th1 / Th17 cells;

[0034] 2) Regulate linoleic acid / bile acid metabolism;

[0035] 3) Regulating intestinal flora, including, for example, increasing the flora combination that is beneficial to inflammatory diseases (such as Ubosiella, Lachnospiraceae_NK4A136_group, Bifidobacterium, Lactobacillus, Limosilatobacillus, Ligilactobacillus, etc.), inhibiting harmful bacteria (such as Ileibacterium, Alistipes, Helicobacter, Faecalibaculum, Turicibacter, etc.);

[0036] 4) Reducing inflammation; preferably, the mechanism of reducing inflammation includes: promoting the production of anti-inflammatory factor IL-10, inhibiting the secretion of inflammatory factors TNF-α, IL-6, IL-8, IL-18, IL-1β and / or reducing the level of intercellular adhesion molecule-1.

[0037] In a fourth aspect, the present invention provides use of Akkermansia muciniphila YG2645 or a derivative thereof as described in the first aspect and the composition as described in the second aspect in the preparation of a product for preventing and / or treating inflammatory diseases.

[0038] In a possible embodiment, the product is a pharmaceutical product.

[0039] In some embodiments, the inflammatory disease is selected from the group consisting of inflammatory bowel disease, irritable bowel syndrome, celiac disease, gastroenteritis, duodenitis, jejunitis, ileitis, peptic ulcer, pouchitis, Collin's ulcer, appendicitis, colitis, diverticular disease, adenocarcinoma, proctitis, mucositis, chemotherapy-related enteritis, radiation-related enteritis, short bowel disease, chronic diarrhea, familial adenomatous polyposis, and perianal fistula. In some embodiments, the inflammatory bowel disease is ulcerative colitis, which may be acute or chronic ulcerative colitis.

[0040] In some embodiments, the Akkermansia muciniphila YG2645 or a derivative thereof or the composition inhibits inflammation by regulating the level of inflammatory factors.

[0041] In particular, in some embodiments of treating inflammatory bowel disease, the Akkermansia muciniphila YG2645 or its derivatives or the composition inhibits inflammation by promoting the production of anti-inflammatory factor IL-10, reducing the levels of inflammatory factors TNF-α, IL-6, IL-8, IL-18, IL-1β and / or reducing the level of intercellular adhesion molecule-1.

[0042] In some embodiments, the medicament is formulated for nasal, oral, or intragastric administration.

[0043] In some embodiments, the drug is in the form of pills, tablets, lozenges, lyophilized powders, granules, capsules, aqueous solutions, alcoholic solutions, oily solutions, syrups, emulsions, suspensions, suppositories, solutions for injection or infusion, ointments, gels, tinctures, creams, patches, lotions, sprays, aerosols, powder sprays, effervescent tablets, transdermal therapeutic systems, microcapsules, implants or sticks.

[0044] In a fifth aspect, the present invention provides a method for preventing and / or treating inflammatory diseases, comprising administering to a subject in need thereof a preventive and / or therapeutically effective amount of Akkermansia muciniphila YG2645 or its derivatives as described in the first aspect or the composition as described in the second aspect.

[0045] In some embodiments, the inflammatory disease is selected from the group consisting of inflammatory bowel disease, irritable bowel syndrome, celiac disease, gastroenteritis, duodenitis, jejunitis, ileitis, peptic ulcer, pouchitis, Collin's ulcer, appendicitis, colitis, diverticular disease, adenocarcinoma, proctitis, mucositis, chemotherapy-related enteritis, radiation-related enteritis, short bowel disease, chronic diarrhea, familial adenomatous polyposis, and perianal fistula. Wherein, the inflammatory bowel disease may be acute or chronic ulcerative colitis.

[0046] The "preventively and / or therapeutically effective amount" may vary depending on the subject of administration, the subject organ, symptoms, the method of administration, etc., and can be determined based on the doctor's judgment, taking into account the type of dosage form, the method of administration, the patient's age and weight, the patient's symptoms, etc.

[0047] Beneficial Effects

[0048] The Akkermansia muciniphila YG2645 of the present invention has the effects of accurately regulating the intestinal-immune-metabolic axis, specifically regulating the intestinal flora or metabolic pathways, enhancing the intestinal barrier function, and alleviating inflammation (for example, cellular inflammation induced by LPS and TNF-α).

[0049] In addition, it was found that YG2645 has a good improvement effect on both acute and chronic UC:

[0050] For acute UC, YG2645 can reduce the expression of TNF-α and IL-6, effectively improve the symptoms of acute UC, and reduce the levels of inflammatory factors.

[0051] For chronic UC, YG2645 intervention can significantly alleviate intestinal epithelial damage and inflammatory infiltration, significantly improve the colon shortening caused by DSS, and reduce colon tissue pathological damage. At the gene level, YG2645 significantly reduced the expression of pro-inflammatory factor IL-18 and upregulated the expression of anti-inflammatory factor IL-10. It showed a good effect in alleviating chronic UC at the phenotypic, pathological and molecular levels.

[0052] In addition, it was found and confirmed that YG2645 can effectively improve acute colitis by regulating Th1 and Th17 cell responses.

[0053] From the above, it can be seen that YG2645 shows great research and application potential, as well as its value as a potential therapeutic strategy. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] One or more embodiments are exemplarily described by the pictures in the accompanying drawings, and these exemplary descriptions do not constitute limitations on the embodiments. The special word "exemplary" here means "used as an example, embodiment or illustrative". Any embodiment described as "exemplary" here is not necessarily interpreted as being superior or better than other embodiments.

[0055] Figure 1 The effect of AKK on the factors related to the cell inflammation model is shown; Figure A shows the protein level of IL-8 in each treatment group, Figure B shows the mRNA level of IL-β in each treatment group, and Figure C shows the ICAM1 level in each treatment group; Compared with the model (TL) group, * p<0.05, ** p<0.01, *** p<0.001; compared with the blank control group (i.e., Con group), # p<0.05, ## p<0.01, ### p<0.001.

[0056] Figure 2 It shows that inactivated AKK improves the functional assessment of acute UC. Among them: Figure A shows the DAI score of inactivated AKK in the 7-day acute UC model; Figure B shows the weight change of inactivated AKK in the 7-day acute UC model; Figure C shows the colon length of inactivated AKK in the 7-day acute UC model; Figure D shows the spleen weight of inactivated AKK in the 7-day acute UC model; Figure E shows the level of colon tumor necrosis factor (TNF-α) in the 7-day acute UC model; Figure F shows the level of pro-inflammatory factor IL-6) in the 7-day acute UC model. Compared with the control group, the model group # p<0.05, ## p<0.01; YG2645 group compared with model group, *p<0.05, ** p<0.01.

[0057] Figure 3 It shows that inactivated YG2645 improves the functional evaluation of chronic UC. Among them: Figure A shows the DAI split of inactivated YG2645 in a 29-day chronic UC model; Figure B shows the weight change of inactivated YG2645 in a 29-day chronic UC model; Figure C shows the colon length of inactivated YG2645 in a 29-day chronic UC model; Figure D shows the anatomy of the colon and spleen of inactivated YG2645 in a 29-day chronic UC model. Compared with the control group, the model group # p<0.05, ## p<0.01; YG2645 group compared with model group, * p<0.05, ** p<0.01.

[0058] Figure 4 It shows that inactivated YG2645 improves chronic UC colon damage and regulates the level of inflammatory factors. Among them: Figure A shows the pathological damage of the colon to the colon by inactivated YG2645 in the 29-day chronic UC model, colon mucosal epithelium (black arrow), inflammatory infiltration (white arrow), basal goblet cells (striped arrow), submucosal edema (scattered arrow), 10× microscope, scale = 100μm; Figure B shows the pathological score of the colon by inactivated YG2645 in the 29-day chronic UC model; Figure C shows the gene level of colon inflammatory factors in the 29-day chronic UC model by inactivated YG2645. Compared with the control group, the model group # p<0.05, ## p<0.01; YG2645 group compared with model group, * p<0.05, ** p<0.01, *** p<0.001.

[0059] Figure 5 Figure 2 shows the results of cell statistics of the reduction of Th1 / Th17 / Th2 / Treg cells in the intestinal mucosa of colitis mice after treatment with inactivated YG2645. Among them: Figure A shows the Th1 cell; mouse colon cell count statistics; Figure B shows the Th17 cell; mouse colon cell count statistics; Figure C shows the Treg cell; mouse colon cell count statistics; Figure D shows the Th2 cell; mouse colon cell count statistics. Error bars represent SD. Compared with the DSS group, * p<0.05, ** p<0.01, *** p<0.001; compared with the blank control group (i.e., Con group), ## p<0.01, ### p<0.001.

[0060] Figure 6 Showing the regulatory effect of YG2645 on intestinal flora. Among them: Figure A is the PCoA analysis of the samples, showing the similarities and differences between different samples; Figure B is the Sobs index analysis, reflecting the species richness; Figure C is the Shannon index analysis, reflecting the species diversity; Figure D is the bacterial community analysis at the phylum level, showing the relative abundance of bacteria in different phyla; Figure E is the bacterial community analysis at the genus level, showing the relative abundance of bacteria in different genera; F is the heat map correlation analysis of intestinal flora and inflammatory markers, red indicates positive correlation, blue indicates negative correlation, and the depth of the color block indicates the strength of the correlation; the error bar indicates the standard deviation (SD).

[0061] Figure 7 Showing that YG2645 alleviates UC through metabolites. Among them: Figure A is the beta diversity analysis of the metabolic profile of colonic contents; Figure B is the volcano map of differential metabolites generated by the comparison between the DSS group and the ND group; Figure C is the volcano map of differential metabolites generated by the comparison between the YG2645 and DSS groups; Figure D is the comparison result of the KEGG enrichment analysis between the YG2645 and DSS groups, identifying the affected metabolic pathways; Figure E is the differential analysis of typical differential metabolites; Figure F is the heat map correlation analysis of typical differential metabolites and intestinal flora; Figure G is the network diagram analysis of metabolites, inflammatory markers and intestinal flora. Error bars represent SD. * p<0.05, **p<0.01, *** p<0.001.

[0062] The Akkermansia muciniphila YG2645 of the present invention is classified and named as Akkermansia muciniphila, with a preservation date of May 10, 2024; the preservation unit is the General Microbiology Center of China Microorganism Culture Collection Administration (referred to as CGMCC, address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code 100101); the preservation number is CGMCC No. 30598. DETAILED DESCRIPTION

[0063] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are 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. Unless otherwise expressly stated, throughout the specification and claims, the term "including" or its variations such as "comprising" or "including" will be understood to include the stated elements or components, without excluding other elements or other components.

[0064] In addition, in order to better illustrate the present invention, numerous specific details are provided in the specific embodiments below. It should be understood by those skilled in the art that the present invention can also be implemented without certain specific details. In some embodiments, raw materials, elements, methods, means, etc. well known to those skilled in the art are not described in detail, so as to highlight the main purpose of the present invention.

[0065] In the following examples, unless otherwise specified, other reagents or raw materials used are commercially available.

[0066] Example 1: Isolation and identification of Akkermansia muciniphila

[0067] (1) Strain isolation

[0068] Feces of healthy children in China were collected, and 1 g of fecal specimens were collected. Fecal genomic DNA was extracted using a fecal genome extraction kit (Qiagen, Germany) according to the operating manual, and its concentration and purity were detected using an enzyme reader. The fecal DNA concentration was adjusted to 10 ng / μL as a qPCR template for detection, and samples with positive test results were isolated for strains.

[0069] Take 1g of positive stool sample and put it into 9mL of PBS buffer. Vortex and mix well. Then dilute it tenfold with sterile saline. -4 , 10 -5 , 10 -6 Three dilution gradients were prepared. 0.5 mL of each dilution was added to a 4.5 mL enrichment tube containing mucin culture medium. The mixture was cultured anaerobically at 37 °C for 72 h. PBS was used for continuous gradient dilution. The dilution was 10 -3 ~10 -6 The diluted liquids were spread on BHI agar medium respectively, and 2 plates were spread for each dilution. The solid plates were cultured in an anaerobic incubator at 37°C for 48 to 72 hours. The colonies were picked and purified on BHI agar medium for more than 3 times until the colonies on the plates were consistent in morphology.

[0070] (2) Strain identification

[0071] A. Colony morphology

[0072] After culturing the 7 isolated strains on BHI agar medium for 24 to 48 hours, the diameter of the colonies ranged from 0.01 to 1.0 mm, and the edges of the colonies were smooth, milky white, and viscous.

[0073] B. 16S rRNA sequencing

[0074] The isolated strains were sent to Sangon Biotech (Shanghai) Co., Ltd. for 16S rRNA gene sequencing, and the 16S rRNA sequencing results were compared with the NCBI database by BLAST (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi). Combined with their colony morphology, the 7 isolated strains were preliminarily determined to be Akkermansia muciniphila (hereinafter referred to as Akk), and were named YG4072, YG2645, YG4071, YG4081, YG2643, YG2639 and YG2609 (hereinafter referred to as 4072, 2645, 4071, 4081, 2643, 2639 and 2609, respectively).

[0075] Through the efficacy screening of the following examples, it was identified that YG2645 has the ability to effectively relieve cellular inflammation and improve acute and chronic ulcerative colitis (UC) and acute colitis, that is, Akkermansia muciniphila YG2645 of the present invention, and its 16S rRNA sequence is shown in SEQ ID NO: 1.

[0076] Example 2: Screening of Akkermansia muciniphila (AKK) strains with anti-inflammatory function based on cell models

[0077] 1.1 Preparation of working solution

[0078] LPS mother solution: weigh 10 mg of LPS powder (Solebol, IL2020-10 mg) into a 15 ml EP tube and add 10 ml of MEM basal medium to prepare a concentrate with a concentration of 1 mg / ml (1000 μg / ml). Sterilize with a 0.22 μm filter membrane, dispense into 1.5 ml EP tubes, and store at -20°C.

[0079] LPS (1 μg / ml) + TNF-α (10 ng / ml) induction reagent: dilute the LPS stock solution with MEM to 1 μg / ml for later use; use LPS (1 μg / ml) solution to dilute tumor necrosis factor α (TNF-α) active protein (Yun-Clone, China) to the required concentration of the experiment (5 ng / ml, 10 ng / ml, 20 ng / ml, 30 ng / ml) for later use.

[0080] Preparation of improved BHI liquid (or solid) culture medium: weigh 0.5 g of L-cysteine ​​hydrochloride solid particles (Solaibao, China), weigh 37 g of BHI powder (oxoid, UK) and add them to 800 mL of distilled water (solid culture medium needs to add 2% agar powder (Solaibao, China), i.e. 20 g), stir and mix well, sterilize at 121°C and high pressure for 15 min, wait until the culture medium temperature drops to 50-60°C, dissolve 5 g of L-threonine (BBI, China) and N-acetylglucose (BBI, China) in 200 mL of distilled water (stir with a glass rod until the solution is completely dissolved), filter with a 0.22 μm filter, add to the high pressure culture medium, and store in a refrigerator at 4°C for use.

[0081] 1.2 Pretreatment of strains

[0082] Resuscitation of strains

[0083] After checking the number of the original strain taken out from -80℃, shake it in a 37℃ water bath to melt it, then add 5% of it to 5mL of modified BHIS liquid culture medium, loosen the bottle cap, and culture it at 37℃ under anaerobic conditions for 15-18h.

[0084] Strain verification

[0085] When the bacterial solution becomes turbid (OD value is about 0.4-0.8), use an inoculation loop to dip into the bacterial solution, draw three lines on the modified BHIS solid culture medium, culture at 37°C under anaerobic conditions for 48-72h, and pick a single colony for mass spectrometry identification.

[0086] Activation of strains

[0087] Pick the correctly identified colonies and put them into 5 mL of modified BHIS liquid culture medium, and culture them at 37°C under anaerobism for 18-22 hours until the bacterial solution becomes turbid; then add 5% of the bacterial solution into new modified BHIS liquid culture medium, and culture them at 37°C under anaerobic conditions for 15 hours until the bacterial solution becomes turbid, and repeat this action twice to achieve the best bacterial activity.

[0088] Dilution of bacterial solution

[0089] Determine the OD value of the cultured AKK bacterial solution and adjust the OD value of the strain to 0.4 (5×10 8 CFU / mL), without centrifugation, take an appropriate amount of bacterial solution and add it to the MEM culture medium required for the experiment, so that its MOI (Multiplicity of infection) = 100, which is used for subsequent cell infection experiments.

[0090] 1.3 Evaluation of the anti-inflammatory ability of AKK strains in inflammatory cell models

[0091] HT-29 cells (Punosai, China) were cultured to the logarithmic phase and the cells were collected. A normal control group (Con), a model group (LPS (1 μg / ml) + TNF-α (10 ng / ml), referred to as TL) and a treatment group (AKK strain) were set up in a 12-well plate. The cell suspension concentration was adjusted to 5×10 5 Each well was inoculated with 1 mL of cell suspension and the cell culture plate was placed in a 37°C, 5% CO2 incubator for 24 h.

[0092] After 24 hours of culture, the cells grew to a monolayer, the culture medium was removed, and the cells were washed once with sterile PBS. 1 ml of MEM culture medium was added to the control and model groups, and 1 mL of AKK strain dilution with an MOI of 100 was added to each well of the treatment group. After 6 hours of infection, the cells were washed once with sterile PBS, 1 mL of MEM culture medium was added to the control group, and 1 mL of induction reagent with a concentration of LPS (1 μg / ml) + TNF-α (10 ng / ml) was added to the model group and the treatment group, respectively. After 2 hours of culture, the cell supernatant was collected, and the protein level of the inflammatory factor IL-8 in the cell supernatant was determined using the MQ60PLUS fully automatic chemiluminescence immunoassay system (Thermogene Bio). TransZol (Quanshijin, R61225) was used to lyse the cells and extract RNA for subsequent RT-PCR, and the gene level content of the inflammatory factors IL-1β and IL-8 and the immunoglobulin superfamily ICAM-1 in the cell supernatant was detected (after obtaining the Ct value, 2 -△△Ct formula for calculation).

[0093] IL-8 is an important indicator in the pathogenesis of UC and a marker for measuring the effectiveness of treatment. Figure 1As shown in Figure A, after induction with LPS (1 μg / ml) and TNF-α (10 ng / ml), the IL-8 level in the cell supernatant increased significantly compared with the control group. However, after treatment with the standard strain ATCC BAA-835 (hereinafter referred to as ATCC835, 835), no significant changes were observed compared with the model group. In contrast, YG4072, YG2645, YG4071, YG4081, YG2643, YG2639, and YG2609 showed better inhibitory effects and significantly reduced the IL-8 factor content. According to the formula: Inhibition rate % = (model group mean - treatment group mean) / model group mean * 100%, the inhibition rates were calculated as follows: ATCC BAA-835 group 31.89%, YG4072 group 34.53%, YG2645 group 62.31%, YG4071 group 54.48%, YG4081 group 83.31%, YG2643 group 62.4%, YG2639 group 67.56%, and YG2609 group 70.12%.

[0094] Further RT-PCR detection showed that the gene expression level of inflammatory factor IL-1β also changed ( Figure 1 This confirmed that strains YG2645, YG2643, and YG2609 had significant effects on inhibiting the expression of IL-1β induced by LPS and TNF-α. Figure 1 As shown in Figure C, after LPS and TNF-α induction, the gene expression level of intercellular adhesion molecule-1 (ICAM-1) increased significantly, while YG2645 treatment significantly reduced the abnormally elevated ICAM-1 level. This indicates that the YG2645 strain can effectively regulate the expression of ICAM-1 induced by LPS and TNF-α.

[0095] In summary, the YG2645 strain can not only significantly inhibit the expression of IL-8 and IL-1β, but also reduce the expression of ICAM-1 associated with inflammation and tumor cell adhesion, thereby effectively alleviating LPS- and TNF-α-induced cellular inflammation. Its anti-inflammatory ability is superior to other strains and has great research and application potential.

[0096] Example 3: Screening of AKK strains with the function of improving acute UC

[0097] (1) Animal experiment design

[0098] 42 8-week-old C57BL / 6J male mice free of specific pathogens (SPF) were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. The mice were raised under a 12-hour light / 12-hour dark cycle, a temperature of 23±3°C, and a humidity of 50%±10%. All mice were allowed to eat and drink freely, and mouse molar sticks were placed in the breeding environment. After one week of adaptive breeding, they were divided into 7 groups, 6 mice in each group, distributed in 2 cages, and acute modeling was carried out for 7 days.

[0099] The specific groupings are:

[0100] (1) Control group: free access to water (0.5% sucrose aqueous solution), intragastric administration of 0.1 ml PBS every day for 7 days;

[0101] (2) Model group: free access to water (0.5% sucrose aqueous solution + 3.0% DSS), intragastric administration of 0.1 ml PBS every day for 7 days;

[0102] (3) YG4081 group: free access to water (0.5% sucrose aqueous solution + 3.0% DSS), oral administration of 0.1 ml 1×10 9 CFUmL -1 Bacterial solution, for 7 days;

[0103] (4) YG2645 group: free access to water (0.5% sucrose aqueous solution + 3.0% DSS), oral administration of 0.1 ml 1×10 9 CFUmL -1 Bacterial solution, for 7 days;

[0104] (5) YG2643 group: free access to water (0.5% sucrose aqueous solution + 3.0% DSS), oral administration of 0.1 ml 1×10 9 CFUmL -1 Bacterial solution, for 7 days;

[0105] (6) YG2609 group: free access to water (0.5% sucrose aqueous solution + 3.0% DSS), oral administration of 0.1 ml 1×10 9 CFUmL -1 Bacterial solution, for 7 days;

[0106] (7) ATCC835 group: free access to water (0.5% sucrose aqueous solution + 3.0% DSS), oral administration of 0.1 ml 1×10 9 CFUmL -1 Bacterial solution for 7 days.

[0107] During the experiment, drinking water for each group was changed every day, DSS (MP, M.W36000-50000) had to be prepared and used immediately, body weight, stool hardness, and fecal occult blood (Shanghai ELISA) were measured every day, and the disease activity index (DAI) was evaluated. On the 7th day of the experiment, the mice were euthanized after blood was collected. The collected blood was centrifuged at 4°C, 5000g for 10 min, the serum was separated, and stored at -80°C. The colon length and spleen weight were recorded, and the colon tissue was collected and fixed with 4% paraformaldehyde. The colon tissue, colon contents, and cecal contents were placed in sterile freeze-drying tubes and quickly frozen with liquid nitrogen, and then stored at -80°C.

[0108] (2) Measurement method

[0109] a. Pathological examination

[0110] The fixed colon tissues were embedded in paraffin, sectioned at 5 mm, stained with hematoxylin and eosin (H&E), and the stained slides were observed under an optical microscope.

[0111] b. Protein detection

[0112] The colon tissue was removed at -80°C, homogenized with an automatic grinder, and the supernatant was used to detect the levels of tumor necrosis factor (TNF-α) and pro-inflammatory factor IL-6 in the colon tissue using an enzyme immunosorbent assay (ELISA) kit.

[0113] Four strains (YG4081, YG2645, YG2643 and YG2609) with anti-inflammatory effects screened in vitro were evaluated for their efficacy in relieving acute ulcerative colitis (UC) in vivo, and the ATCC835 group was used as the standard control bacteria group. The disease activity index (DAI) score comprehensively considers the weight changes, stool characteristics and fecal occult blood levels of mice, and can phenotypically reflect the progression of acute UC. During the seven-day modeling experiment, it was observed that the DAI score of the model group increased significantly and the body weight decreased significantly compared with the control group. YG2645 treatment significantly reduced the DAI score and significantly alleviated the weight loss caused by DSS ( Figure 2 Figures A and B in the middle). On the seventh day of modeling, the mice were dissected and found that the colon length of the model group was significantly shortened and the spleen weight was significantly increased compared with the control group. YG2645 treatment can significantly alleviate the shortening of colon length and increase of spleen weight caused by DSS ( Figure 2 In addition, the expression of inflammatory factors in colon tissue was detected.

[0114] The results showed that compared with the control group, the levels of tumor necrosis factor (TNF-α) and pro-inflammatory factor IL-6 in the model group were increased, while YG2645 was able to reduce the expression of TNF-α and IL-6, among which the decrease in TNF-α showed a significant difference ( Figure 2The above results indicate that YG2645 can effectively improve the symptoms of acute UC and reduce the levels of inflammatory factors, demonstrating its value as a potential therapeutic strategy.

[0115] Example 4: YG2645 precisely regulates the intestinal-immune-metabolism axis to significantly improve chronic UC

[0116] 4.1YG2645 improves chronic UC and intestinal epithelial barrier damage

[0117] (1) Animal experiment design

[0118] Eighteen 8-week-old C57BL / 6J male mice free of specific pathogens (SPF) were purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. The mice were raised under a 12-hour light / 12-hour dark cycle, a temperature of 23±3°C, and a humidity of 50%±10%. All mice were allowed to eat and drink freely, and mouse molar sticks were placed in the feeding environment. After one week of adaptive feeding, they were divided into 7 groups, 6 mice in each group, distributed in 2 cages, and acute modeling was established for 29 days.

[0119] The specific groupings are:

[0120] 1) Control group: gavage with 0.1 ml PBS every day for 29 days;

[0121] 2) Model group: gavage with 0.1 ml PBS every day for 29 days;

[0122] 3) YG2645 group: 0.1 ml 1×10 9 CFUmL -1 Bacterial solution, lasting 29 days.

[0123] The control group freely drank 0.5% sucrose aqueous solution throughout the whole process; the model group and YG2645 group freely drank (0.5% sucrose aqueous solution + 2.5% DSS) for 5 days and (0.5% sucrose aqueous solution) for 7 days, 5 days + 7 days for 1 round, a total of 2.5 rounds. During the experiment, the drinking water of each group was changed every day, and DSS (MP, M.W36000-50000) needed to be prepared and used immediately. The body weight, stool hardness and fecal occult blood (Shanghai ELISA, ml095102) were measured every day, and the disease activity index (DAI) was evaluated. On the 29th day of the experiment, the mice were euthanized after blood was collected. The collected blood was centrifuged at 4°C, 5000g for 10 minutes, serum was separated, and stored at -80°C. The colon length and spleen weight were recorded, and the colon tissue was collected and fixed with 4% paraformaldehyde. The colon tissue, colon contents and cecal contents were placed in sterile freeze-drying tubes and quickly frozen with liquid nitrogen and stored at -80°C.

[0124] (2) Measurement method

[0125] a. Pathological examination

[0126] The fixed colon tissues were embedded in paraffin, sectioned at 5 mm, stained with hematoxylin and eosin (H&E) and PAS, and the stained slides were observed under a light microscope.

[0127] b. Genetic testing

[0128] Colon tissue was taken out at -80℃, and RNA was extracted from colon tissue using RNA extraction kit (TAKARA), and RNA concentration and purity were determined. RNA samples were reverse transcribed using TaKaRa kit and manufacturer's instructions. Finally, qPCR kit (Tiangen) and fluorescence quantitative PCR amplification instrument were used. The expression level of each gene was detected.

[0129] (3) Experimental results

[0130] After achieving satisfactory results in the acute ulcerative colitis (UC) model, we continued to evaluate the effect of YG2645 in the chronic UC model to explore its role in different UC disease courses.

[0131] The results showed that YG2645 could significantly reduce the disease activity index (DAI) score and improve the weight loss caused by DSS ( Figure 3 The anatomical results showed that 2.5% DSS intervention could lead to shortening of colon length, while YG2645 intervention significantly improved the shortening of colon caused by DSS ( Figure 3 Histopathological examination further confirmed the efficacy of YG2645.

[0132] HE staining and PAS staining results showed that DSS induced damage to the colonic mucosal epithelial structure, inflammatory infiltration, reduction of basal goblet cells, and edema of the submucosal layer. YG2645 intervention can significantly alleviate intestinal epithelial damage and inflammatory infiltration, and reduce colon tissue pathological damage ( Figure 4 In addition, at the gene level, YG2645 significantly reduced the expression of the pro-inflammatory factor IL-18 and upregulated the expression of the anti-inflammatory factor IL-10 ( Figure 4 (Figure C).

[0133] In summary, YG2645 showed good effects in alleviating chronic UC at the phenotypic, pathological and molecular levels. It can effectively improve the shortening of colon length, reduce intestinal tissue damage, and regulate the expression of inflammatory factors, showing its potential in the treatment of chronic UC.

[0134] 4.2YG2645 regulates cellular immune response through Th1 / Th17

[0135] (1) Colonic lamina propria cell extraction

[0136] a. Take a section of intestine and quickly place it in sterile pre-cooled ice HBSS (w / o), keeping the whole operation on ice; soak it in ice HBSS (w / o) and carefully clean the mesentery and fat, and try to clean it as clean as possible to avoid affecting the extraction effect.

[0137] b. Cut the intestine longitudinally, clean the intestinal contents and mucus, keep the intestine moist, and then cut it into 0.5 cm segments.

[0138] c. Add 20 mL of pre-digestion solution to each 50 mL centrifuge tube and transfer the intestinal segments to the tube.

[0139] d. Rotate on a rotator and incubate at 37°C for 20 mins. Vortex for 10 s and filter with a 100 μm filter.

[0140] e. Transfer the intestinal lamina propria tissue on the filter to a 50mL centrifuge tube containing 20mL pre-digestion solution, rotate on a rotator, and incubate at 37℃ for 20mins. At the same time, prepare: add 2.35mL digestion solution (w) to each C tube, add 162.5μL enzyme MIX solution to the C tube containing 2.35mL digestion solution, and mix gently. Preheat at 37℃ for 15mins.

[0141] f. Vortex for 10 seconds, filter with a 100 μm filter, transfer the intestinal tissue to a C tube containing digestive enzyme MIX, and tighten.

[0142] g. Incubate on a rotator at 37°C, 100 rpm for 20 mins.

[0143] h. Tighten the C tube and invert it in the GentleMACS Dissociator. Make sure the intestinal tissue is in the inverted bottom liquid and select GentleMACSprogramm_intestine_01.

[0144] i. Immediately after the procedure, remove and add 5 mL of PB buffer and filter with a 100 μm filter.

[0145] j. Rinse the filter and tube C with 10 mL of PB buffer.

[0146] k. Centrifuge at 300×g for 10 min at room temperature. The precipitate is relatively loose at this time. Aspirate the supernatant as completely as possible and resuspend the obtained cells in 4 mL of 40% percoll solution.

[0147] 1. Add 4 mL of 80% Percoll solution to a 15 mL centrifuge tube, and slowly add the resuspended liquid from step k to cover the 80% Percoll solution. Centrifuge at room temperature for 20 minutes at 600 g (1 for increase and 0 for decrease). After centrifugation, obvious stratification is observed, and the white flocculent layer in the middle is the colonic lamina propria cells. Carefully aspirate and transfer to a new 15 mL centrifuge tube, add iced HBSS (w / o) to make up to 10 mL, centrifuge at 4°C, 5000 rpm for 10 minutes, and gently pour off the supernatant.

[0148] *Solution dilution and preparation requires sterile operation

[0149] Prepare 50 mL of 1640 culture medium containing the cell stimulator GolgiPlug (add 2 μL of GolgiPlug to every 1 mL of 1640 complete culture medium).

[0150] m. Resuspend the cells with the remaining liquid at the bottom and add them to the prepared 1640 medium containing stimulation solution and stimulate at 37℃ for 5 hours.

[0151] (2) Antibody staining

[0152] a. Gently blow the cells in the well plate and transfer them to a 1.5 mL EP tube. Centrifuge at 300×g for 5 minutes at 4°C, remove the supernatant, resuspend in 50 μL CD16 / 32 blocking solution (50 μL PBS plus 2 μL CD16 / 32), and incubate on ice for 10 minutes.

[0153] b. Prepare a cell surface antibody mixture with PBS, cell death antibody dye, CD45, CD3, CD4, and CD8 antibodies to a final volume of 50 μL, and vortex to mix; add 50 μL of the surface antibody mixture to the liquid obtained in step a, and incubate on ice for 30 minutes away from light.

[0154] c. After incubation, add 1 mL of PBS and centrifuge at 300×g for 5 minutes at 4°C. Remove the supernatant and resuspend the cells with the remaining liquid at the bottom.

[0155] d. Add 200 μL Fixation / Permeabilization solution to each tube to resuspend the cells (including negative control and single-stained tubes, both need to be fixed and permeated), resuspend thoroughly (blow several times with a 1 mL gun), and incubate on ice for 40 minutes away from light.

[0156] *Use 1× Permeabilization buffer to prepare IFN-γ and IL-17A antibody suspension to a final volume of 50 μL / tube.

[0157] e. Add 200 μL 1× Permeabilization buffer and centrifuge at 600×g at 4°C for 5 minutes.

[0158] f. Remove the supernatant and resuspend the cells with the remaining liquid at the bottom. Resuspend the cells with 50 μL of cytokine staining solution and vortex gently. Incubate at 4°C for 30 minutes in the dark.

[0159] g. Add 200 μL 1× Permeabilization buffer to each tube and centrifuge at 600×g at 4°C.

[0160] h. After removing the supernatant, resuspend the cell pellet with 200 μL 1× Permeabilization solution and store at 4°C in the dark until ready for loading.

[0161] (3) Results analysis

[0162] The pathogenesis of ulcerative colitis (UC) is closely related to four immune cell subsets: Th1, Th2, Th17, and Treg. Among them, the enhanced activity of Th1 cells and the involvement of Th2 cells may lead to aggravated intestinal inflammation. IFN-γ and IL-12 produced by Th1 cells can aggravate intestinal inflammation, leading to intestinal mucosal damage and ulcer formation. IL-4, IL-5, and IL-13 produced by Th2 cells may promote excessive secretion of intestinal mucus, increase intestinal permeability, and further aggravate the inflammatory response. Th17 cells also play a key role in the inflammatory response. The increase in their number indicates that the inflammatory response is ongoing. IL-17 produced by Th17 cells can recruit and activate inflammatory cells such as neutrophils, leading to worsening intestinal inflammation. In contrast, Treg cells play a regulatory role in the immune system. They secrete cytokines such as IL-10 and TGF-β to inhibit the immune response and prevent excessive inflammation.

[0163] In this study, flow cytometry was used to analyze the effect of inactivated YG2645 on the distribution of immune cell subsets. Figure 5 As shown in Figures AD, inactivated YG2645 significantly downregulated the generation of Th1 and Th17 cells in the intestinal mucosa of mice with DSS-induced colitis, with specific inhibition rates of Th1: 68.58% and Th17: 68.66%, respectively; while it had no significant effect on Th2 and Treg cells.

[0164] These results suggest that YG2645 can effectively improve acute colitis by regulating Th1 and Th17 cell responses. In addition, YG2645 strain is a potential probiotic preparation with economic benefits in the treatment of colitis and deserves further development and research.

[0165] 4.3 Regulatory effect of YG2645 on intestinal flora

[0166] The 16S rRNA gene sequencing technology was used to explore the effect of YG2645 on the intestinal flora of DSS-induced colitis. Figure 6 As shown in Figures B and C, YG2645 increased the number of gut-specific OTUs and improved the Sobs index compared with the DSS group. The Shannon index showed that YG2645 increased the alpha diversity of the intestinal flora. To further understand the role of microbial diversity, beta diversity was analyzed by principal coordinate analysis (PCoA; Bray-Curtis distance). The NC, DSS, and YG2645 groups showed different clustering patterns on the PCoA plot, indicating significant differences among the three groups. YG2645 significantly altered the beta diversity of the intestinal flora ( Figure 6 At the phylum level, compared with the DSS group, YG2645 intervention increased the relative abundance of Firmicutes, Actinobacteria, and Proteobacteria, while decreasing the relative abundance of Verrucomicrobia and Bacteroidetes ( Figure 6 At the genus level, YG2645 increased the abundance of beneficial bacteria such as Dubosiella, Lachnospiraceae_NK4A136_group, Bifidobacterium, Lactobacillus, Limosilatobacillus, and Ligilactobacillus, and decreased the abundance of harmful bacteria such as Ileibacterium, Alistipes, Helicobacter, Faecalibaculum, and Turicibacter ( Figure 6 Figure E). By analyzing the relationship between changes in intestinal microbial composition and colitis-related indicators, it was found that these beneficial bacteria were negatively correlated with DAI, inflammation score, IL-6, IL-8, IL-18, Th17 and Th1, and positively correlated with colon length and anti-inflammatory factor IL-10. In contrast, the correlation trend between harmful bacteria and colitis-related indicators was the opposite ( Figure 6 (Fig. F).

[0167] 4.4YG2645 alleviates UC by regulating linoleic acid and hyobolic acid

[0168] To further explore the effect of YG2645 on the fecal metabolome of DSS-induced mice, the differences in fecal metabolomes among different groups were analyzed. Partial least squares discriminant analysis (PLS-DA) is a supervised machine learning algorithm that can distinguish the overall variability of samples.

[0169] The PLS-DA visualization results showed that the NC, DSS and YG2645 groups showed an obvious clustering trend ( Figure 7Figure A), indicating that YG2645 intervention significantly affected the fecal metabolome of DSS-induced mice. Compared with the ND group, 217 metabolites were increased and 192 metabolites were decreased in the DSS group; while compared with the DSS group, 76 metabolites were increased and 80 metabolites were decreased in the YG2645 group ( Figure 7 Figures B and C).

[0170] In order to analyze the mechanism of sample metabolic changes, metabolic pathway enrichment analysis was performed based on differential metabolites. The results showed that 11 metabolic pathways were significantly enriched in the YG2645 group compared with the DSS group ( Figure 7 In addition, 17 differential metabolites were screened between the two groups (P<0.05, VIP>1), among which the levels of linoleic acid, liquiritigenin, taurocholic acid, hyocholic acid and coumarin increased, while the levels of 12,13-EpOME, docosanedioate, 9R,10S-EpOME and tetrahydrocortisol decreased in the YG2645 group ( Figure 7 By analyzing the relationship between changes in intestinal metabolite composition and the abundance of microbial OTUs, it was found that at the genus level, the contents of linoleic acid, glycyrrhizin, taurocholic acid, and hyocholic acid were significantly positively correlated with Lachnospiraceae_NK4A136_group, Bifidobacterium, lactobacillus, Limosilatobacillus, and Ligilactobacillus, but significantly negatively correlated with Ileibacterium, Helicobacter, Faecalibaculum, and Turicibacter ( Figure 7 (Fig. F).

[0171] To explore the mechanism of YG2645 in alleviating colitis in mice, Pearson correlation analysis was performed to analyze the relationship between differential bacterial genera, key metabolites and inflammatory indicators. The results showed that linoleic acid and hyobolic acid were significantly positively correlated with Lachnospiraceae_NK4A136_group, Limosilatobacillus and Ligilactobacillus, and significantly negatively correlated with Th17, inflammatory score and disease activity index (DAI) ( Figure 7 Therefore, it is inferred that linoleic acid and hyocholic acid may be the key metabolites of YG2645 in alleviating colitis.

[0172] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A strain of Akkermansia muciniphila YG2645 or a derivative thereof, characterized in that: The Akkermansia muciniphila YG2645 is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration, with a deposit date of May 10, 2024 and a deposit number of CGMCC No.30598.

2. The Akkermansia muciniphila YG2645 or a derivative thereof according to claim 1, characterized in that The 16SrRNA gene sequence of Akkermansia muciniphila YG2645 is shown in SEQ ID NO:

1.

3. Akkermansia muciniphila YG2645 or a derivative thereof according to claim 1 or 2, characterized in that The derivative is the inactivated bacteria, extract, culture, culture supernatant, fermentation product, fermentation supernatant or a combination thereof of the Akkermansia muciniphila YG2645.

4. A composition, characterized in that The composition comprises an effective amount of a first active component, wherein the first active component comprises Akkermansia muciniphila YG2645 or a derivative thereof according to any one of claims 1 to 3.

5. The composition according to claim 4, characterized in that The composition is a pharmaceutical composition, a food composition or a feed composition.

6. The composition according to claim 4 or 5, characterized in that The composition further comprises a second active ingredient.

7. The composition according to claim 6, characterized in that The second active component comprises a probiotic, a postbiotic, a prebiotic, an antimicrobial agent, an immunomodulator, an anticancer agent, an inflammatory therapeutic agent, or a combination thereof.

8. Use of Akkermansia muciniphila YG2645 or a derivative thereof according to any one of claims 1 to 3 or a composition according to any one of claims 4 to 7 in the preparation of a product, preferably a pharmaceutical product, for the following purposes: 1) Regulate the balance of Th1 / Th17 cells; 2) Regulate linoleic acid / bile acid metabolism; 3) Regulate intestinal flora; 4) reducing inflammation; preferably, the mechanism of reducing inflammation includes: It promotes the production of anti-inflammatory factor IL-10, inhibits the secretion of inflammatory factors TNF-α, IL-6, IL-8, IL-18, IL-1β and / or reduces the level of intercellular adhesion molecule-1.

9. Use of Akkermansia muciniphila YG2645 or a derivative thereof according to any one of claims 1 to 3 or a composition according to any one of claims 4 to 7 in the preparation of a product for preventing and / or treating inflammatory diseases.

10. The use according to claim 9, characterized in that The product in question is a pharmaceutical product; and / or, the inflammatory disease is selected from the group consisting of inflammatory bowel disease, irritable bowel syndrome, celiac disease, gastroenteritis, duodenitis, jejunitis, ileitis, peptic ulcer, pouchitis, Collin's ulcer, appendicitis, colitis, diverticular disease, adenocarcinoma, proctitis, mucositis, chemotherapy-related enteritis, radiation-related enteritis, short bowel disease, chronic diarrhea, familial adenomatous polyposis, and perianal fistula; Preferably, the inflammatory bowel disease is ulcerative colitis.

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