Bifidobacterium breve BBr26 capable of inhibiting hyaluronidase activity and allergy relief, and its application, product and method

Abstract

The present invention provides a Bifidobacterium breve BBr26 with the ability to inhibit hyaluronidase activity and relieve allergies, and its application, product and method, which belong to the field of microbial technology. The present invention provides a Bifidobacterium breve ( Bifidobacterium breve ) strain BBr26, with a deposit number of CGMCC No. 32424. The present invention also provides the use of this strain, Bifidobacterium breve BBr26, in the preparation of atopic dermatitis-relieving pharmaceuticals, immune-enhancing foods, and / or antibacterial agents, as well as products and methods based on this strain. The strain of the present invention exhibits broad-spectrum antibacterial properties, can alleviate allergies and atopic dermatitis, and effectively regulates the immune system.

Description

Technical Field

[0001] The present invention belongs to the field of microbial technology, and specifically relates to Bifidobacterium breve BBr26 capable of inhibiting hyaluronidase activity and allergy relief, as well as applications, products and methods thereof. Background Art

[0002] The prevalence of allergic diseases has been increasing in recent years. Common allergic diseases include allergic rhinitis, allergic asthma, allergic eczema, food allergies, atopic dermatitis, and contact dermatitis. Allergic diseases are often accompanied by inflammation. Allergic reactions can be categorized by their mechanisms: Type I, II, III, and IV. Each type of allergic reaction has distinct mechanisms. In type I allergic reactions, IgE antibodies, following contact with a specific antigen, bind to IgE receptors on mast cells and basophils. This bridging action triggers rapid degranulation and the release of pharmacological mediators of local inflammation, resulting in clinical symptoms. In type II allergic reactions, antibodies bind to cell surface antigens or to antigens or haptens adsorbed on the cell surface, leading to complement involvement and cell and tissue damage. In type III allergic reactions, complement activation and the release of lytic enzymes by neutrophils cause tissue damage. Inhaled antigens can cause localized lung damage. Systemic immune complexes formed by antibodies and microorganisms or self-antigens lead to intravascular deposition of immune complexes, causing vasculitis or glomerulonephritis. Type IV allergic reaction antigens sensitize the body, forming inflammatory T cells and sensitized T cells, which again combine with specific antigens to directly kill target cells or produce a variety of cytokines, resulting in inflammation or delayed hypersensitivity reactions.

[0003] Hyaluronidase is a hydrolase that degrades hyaluronic acid, a major component of the extracellular matrix of connective tissue. It is present in most IgE-mediated type I and T-cell-mediated type IV allergic reactions. The vast majority of allergic diseases are type I allergic reactions associated with hyaluronidase activity in vivo. Therefore, the in vitro hyaluronidase inhibition assay is an effective method for rapid screening of anti-allergic substances in vitro. Simultaneously, inhibiting hyaluronidase activity ensures normal hyaluronic acid content and function. Hyaluronidase is clinically used as a drug diffusion agent, particularly in tumor treatment and subcutaneous drug delivery, and holds great promise.

[0004] Bifidobacterium breve BBr26 is a Gram-positive anaerobic bacterium commonly found in infant feces and belongs to the subspecies Bifidobacterium. However, there are no reports of Bifidobacterium breve being able to alleviate allergic symptoms by inhibiting hyaluronidase.

[0005] Therefore, there is a need in the art to develop a new strain of Bifidobacterium breve that simultaneously inhibits hyaluronidase activity, has anti-allergic effects, and has broad-spectrum antibacterial properties. Summary of the Invention

[0006] To address the above-mentioned needs in the art, the present invention provides a product and method for alleviating atopic dermatitis using a strain of Bifidobacterium breve BBr26.

[0007] The purpose of the present invention is achieved through the following technical solutions:

[0008] A Bifidobacterium breve that inhibits hyaluronidase activity and relieves allergies ( Bifidobacterium breve )BBr26, its deposit number is CGMCC NO.32424.

[0009] The preservation number is CGMCC NO.32424 Bifidobacterium breve ( Bifidobacterium breve ) BBr26, application in the preparation of medicines for relieving atopic dermatitis, and / or foods for enhancing immune function, and / or antibacterial agents.

[0010] The relief of atopic dermatitis includes: reducing ear thickness, reducing skin thickness, relieving skin inflammatory infiltration, reducing mast cell count, increasing thymus index, reducing spleen index, inhibiting hyaluronidase activity, and regulating cytokine levels;

[0011] Preferably, the immunity enhancement includes: increasing the proliferation rate of immune cells RAW264.7, promoting the secretion of INF-γ by spleen cells, inhibiting the secretion of tryptase by mast cells, and regulating cytokine levels;

[0012] Preferably, regulating cytokine levels includes: reducing the levels of IL-4 and IL-13, and increasing the levels of IL-10 and INF-γ;

[0013] Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, and Propionibacterium acnes.

[0014] A drug for relieving atopic dermatitis, comprising: an active ingredient; the active ingredient comprises: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0015] The drug for relieving atopic dermatitis further comprises: excipients.

[0016] A food for enhancing immune function, comprising: active ingredients; the active ingredients comprising: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0017] The food for enhancing immune function further comprises auxiliary materials.

[0018] An antibacterial agent, comprising: an active ingredient; the active ingredient comprises: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0019] The antibacterial agent further comprises: auxiliary materials.

[0020] An in vitro antibacterial method using Bifidobacterium breve (CGMCC NO.32424) Bifidobacterium breve ) BBr26 is antibacterial.

[0021] The beneficial effects of the present invention are as follows:

[0022] The present invention screened a new strain from infant fecal samples, and molecular identification showed that the strain was Bifidobacterium breve ( Bifidobacterium breve ), and named it strain BBr26. The present invention has been confirmed through a large number of experiments that the new strain BBr26 has a high tolerance to gastric juice and can relieve polytype allergic reactions. The strain BBr26 can effectively inhibit the activity of hyaluronidase, inhibit the secretion of tryptase by mast cells, promote the secretion of INF-γ by mouse spleen cells, and relieve atopic dermatitis in mice. At the same time, the strain BBr26 also has a significant effect on the proliferation rate of macrophages RAW264.7, effectively regulating the proliferation of macrophages RAW264. to perform immunomodulation. Not only that, the strain BBr26 of the present invention also has a strong and broad-spectrum antibacterial function, and can play a good inhibitory effect on many pathogens including Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, and Propionibacterium acnes.

[0023] Bifidobacterium breve of the present invention ( Bifidobacterium breve ) The deposit information of BBr26 is as follows:

[0024] Deposit number: CGMCC NO.32424;

[0025] Classification name: Bifidobacterium breve Bifidobacterium breve ;

[0026] Deposit date: October 30, 2024;

[0027] Depository: China General Microbiological Culture Collection Center;

[0028] Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1This is a bar graph showing the results of the assay of the activity of Bifidobacterium breve BBr26 in inhibiting tryptase secretion from mast cells in Experimental Example 3 of the present invention.

[0030] Figure 2 This is a line graph showing the changes in ear thickness in each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0031] Figure 3 These are skin tissue sections of each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0032] Figure 4 This is a microscopic examination of mast cell staining in atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0033] Figure 5 This is a bar graph of the thymus index of each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0034] Figure 6 This is a bar graph of the spleen index of each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0035] Figure 7 This is a bar graph showing the levels of IL-4, IL-13, IL-10, and INF-γ in the serum of mice in each group of the atopic dermatitis model of Experimental Example 8 of the present invention.

[0036] Figure 8 This is a bar graph showing the serum IgE levels in each group of mice in the atopic dermatitis model of Experimental Example 8 of the present invention.

[0037] Figure 1 The meanings of each mark are listed as follows: CK: blank control group; C48 / 80: positive drug control group; BBr26: probiotic treatment group.

[0038] Figures 2 - 8 The meanings of each mark are listed as follows: NC: blank control group; MC: model group; Keto: positive drug intervention group; BBr26: Bifidobacterium breve BBr26 intervention group. DETAILED DESCRIPTION

[0039] The details of the present invention are further described below in conjunction with specific embodiments and experimental examples, but the scope of protection of the present invention is not limited thereto.

[0040] Sources of biological materials

[0041] 1. The Bifidobacterium breve M-16V used in Experimental Example 2 of the present invention is commercially available;

[0042] 2. Experimental Example 3 of the Present Invention Mast cells were purchased from China Center for Type Culture Collection.

[0043] 3. The Balb / c mice used in Experimental Example 4 of the present invention were purchased from Hubei Safety Evaluation Center.

[0044] 4. The Escherichia coli ATCC25922, Salmonella Enteritidis ATCC14028, and Staphylococcus aureus ATCC25923 used in Experimental Example 6 of the present invention were purchased from the Guangdong Microorganism Collection Center; Streptococcus pasteurianus was Streptococcus pasteurianus 12758, which has been registered in NCBI with the accession number MW445225; Streptococcus Parisi was Streptococcus Parisi 21178, which has been registered in NCBI with the accession number OM403351; and Streptococcus anginosus was Streptococcus pharyngitis strain 4997, which has been registered in NCBI with the accession number MT459365. They are currently maintained in the applicant's laboratory. Staphylococcus saprophyticus and Propionibacterium acnes are maintained in the Wuhan Weikang Probiotics Research Institute Co., Ltd. The applicant promises to release these to the public for use in verifying the technical effects of the present invention within 20 years from the filing date.

[0045] 5. The macrophage RAW264.7 used in Experimental Example 7 of the present invention is commercially available.

[0046] 6. The Balb / c mice used in Experimental Example 8 of the present invention were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd.

[0047] The first group of examples, Bifidobacterium breve BBr26 of the present invention

[0048] This group of embodiments provides a method for inhibiting hyaluronidase activity and relieving allergies. Bifidobacterium breve ) BBr26. All embodiments in this group have the following common features: the Bifidobacterium breve ( Bifidobacterium breve ) The deposit number of BBr26 is CGMCC NO.32424.

[0049] Any act of culturing, propagating, fermenting, enriching, producing, preparing, using, inoculating, amplifying, transforming, modifying, transforming, selling, or offering for sale the strain of Bifidobacterium breve BBr26 with the deposit number CGMCC NO. 32424, and / or using the strain of Bifidobacterium breve BBr26 with the deposit number CGMCC NO. 32424 in combination with other probiotics, and / or using the strain of Bifidobacterium breve BBr26 with the deposit number CGMCC NO. 32424 to antagonize various pathogens, and / or prepare antibacterial agents, and / or prepare foods that enhance immune function, and / or foods that regulate intestinal flora, all fall within the scope of protection of the present invention.

[0050] The other probiotics include but are not limited to Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii lactis subsp. lactis, Lactobacillus helveticus, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus salivarius, Saccharomyces cerevisiae, Torulella delbrueckii, Candida, Wickham's yeast, Pichia pastoris, Saccharomyces boulardii, Torulopsis spp., Schizosaccharomyces spp., Rhodotorula spp., and Schizosaccharomyces pombe. , Saccharomyces boulardii, Bacillus thuringiensis, Brevibacillus laterosporus, Bacillus megaterium, Bacillus mucilaginosus, Bacillus azotobacterium, Bacillus sphaericus, Clostridium butyricum, Bifidobacterium adolescentis, Bifidobacterium angularis, Bifidobacterium animalis, Bifidobacterium asteroides, Bifidobacterium bifidum, Bifidobacterium bovis, Bifidobacterium breve, Bifidobacterium dentate, Bifidobacterium infantis (i.e., Bifidobacterium longum infantis subsp. infantis), Bifidobacterium lactis (i.e., Bifidobacterium animalis subsp. cremoris), Bifidobacterium longum, Bifidobacterium pseudocatenulatum, as well as Bifidobacterium thermophilum and Bifidobacterium acidothermophilum.

[0051] Those skilled in the art can, based on actual production needs, combine conventional technical means or basic common sense of pharmaceutical production processes (for example, "Encyclopedia of Preparation Technology", "Encyclopedia of Food and Food Production", "Lactic Acid Bacteria and Fermented Foods", "Research and Application of Microbial Agent Technology", etc.) to routinely select or adjust pharmaceutical excipients, and then prepare Bifidobacterium breve BBr26 with a preservation number of CGMCC NO. 32424 into products with different dosage forms, different storage conditions, and different shelf lives. This is possible and easy to do for those skilled in the art without any technical obstacles.

[0052] Group 2 Example: Application of Bifidobacterium breve BBr26 of the present invention

[0053] This group of examples provides a collection number of CGMCC NO.32424 Bifidobacterium breve ( Bifidobacterium breve ) BBr26, application in the preparation of medicines for relieving atopic dermatitis, and / or foods for enhancing immune function, and / or antibacterial agents.

[0054] In a specific embodiment, the relief of atopic dermatitis includes: reducing ear thickness, reducing skin thickness, relieving skin inflammatory infiltration, reducing the number of mast cells, increasing thymus index, reducing spleen index, inhibiting hyaluronidase activity, and regulating cytokine levels;

[0055] Preferably, the immunity enhancement includes: increasing the proliferation rate of immune cells RAW264.7, promoting the secretion of INF-γ by spleen cells, inhibiting the secretion of tryptase by mast cells, and regulating cytokine levels;

[0056] Preferably, regulating cytokine levels includes: reducing the levels of IL-4 and IL-13, and increasing the levels of IL-10 and INF-γ;

[0057] Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, and Propionibacterium acnes.

[0058] The third group of examples, the drug for relieving atopic dermatitis of the present invention

[0059] This group of embodiments provides a drug for relieving atopic dermatitis. All embodiments in this group have the following common features: the drug for relieving atopic dermatitis comprises: an active ingredient; the active ingredient comprises: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0060] In a further embodiment, the drug for relieving atopic dermatitis is characterized in that it further comprises: excipients.

[0061] In a more specific embodiment, the excipients are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0062] According to the present invention, based on the different needs of actual production applications, combined with conventional technical means in the field of formulation preparation (for example, "Encyclopedia of Formulation Technology", "Encyclopedia of Food and Food Production", "Lactic Acid Bacteria and Fermented Foods", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare Bifidobacterium breve BBr26 with a deposit number of CGMCC NO. 32424 into different dosage forms, such as powders, tablets, oral liquids, gels, patches, sprays, lotions, granules, etc.

[0063] In a specific embodiment, the dosage form of the drug for relieving atopic dermatitis is selected from: one or more of powder, tablet, liquid, and capsule.

[0064] The fourth group of embodiments, the immune function enhancing food of the present invention

[0065] This group of embodiments provides a food for enhancing immune function. All embodiments of this group have the following common features: the food for enhancing immune function comprises: active ingredients; the active ingredients include: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0066] In a further embodiment, the immune function enhancing food further includes: auxiliary materials.

[0067] In a more specific embodiment, the excipients are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0068] According to the present invention, based on the different needs of actual production applications, combined with conventional technical means in the field of formulation preparation (for example, "Encyclopedia of Formulation Technology", "Encyclopedia of Food and Food Production", "Lactic Acid Bacteria and Fermented Foods", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare Bifidobacterium breve BBr26 with a deposit number of CGMCC NO. 32424 into different dosage forms, such as powders, tablets, oral liquids, gels, patches, sprays, lotions, granules, etc.

[0069] In a specific embodiment, the dosage form of the immune-enhancing food is selected from: one or more of powder, tablet, liquid, and capsule.

[0070] The fifth group of examples, the antibacterial agent of the present invention

[0071] This group of embodiments provides an antibacterial agent. All embodiments of this group have the following common features: the antibacterial agent comprises: an active ingredient; the active ingredient comprises: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0072] In a further embodiment, the antibacterial agent further comprises: an auxiliary material.

[0073] In a more specific embodiment, the excipients are selected from: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0074] According to the present invention, based on the different needs of actual production applications, combined with conventional technical means in the field of formulation preparation (for example, "Encyclopedia of Formulation Technology", "Encyclopedia of Food and Food Production", "Lactic Acid Bacteria and Fermented Foods", "Research and Application of Microbial Agent Technology", etc.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare Bifidobacterium breve BBr26 with a deposit number of CGMCC NO. 32424 into different dosage forms, such as powders, tablets, oral liquids, gels, patches, sprays, lotions, granules, etc.

[0075] In a specific embodiment, the dosage form of the antibacterial agent is selected from: one or more of powder, tablet, liquid, and capsule.

[0076] The sixth group of examples, the in vitro antibacterial method of the present invention

[0077] This group of embodiments provides an in vitro antibacterial method. All embodiments in this group have the following common features: using Bifidobacterium breve ( Bifidobacterium breve ) BBr26 is antibacterial.

[0078] In a specific embodiment, the antibacterial spectrum includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, and Propionibacterium acnes.

[0079] “Above” and “below” in this article include the original number.

[0080] In this article, "Bifidobacterium breve ( Bifidobacterium breve )BBr26", "Bifidobacterium breve ( Bifidobacterium breve ) strain BBr26", "Bifidobacterium breve BBr26", "BBr26", "BBr26 strain", "strain BBr26" all refer to: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0081] The culture medium and reagents used in the experimental examples of the present invention are as follows:

[0082] MRS medium: peptone 10 g / L, beef extract powder 5 g / L, yeast powder 5 g / L, glucose 20 g / L, anhydrous sodium acetate 5 g / L, diammonium hydrogen citrate 2 g / L, Tween 80 1 g / L, K2HPO4 2 g / L, MgSO4 0.2 g / L, MnSO4 0.05 g / L, pH = 6.80 ± 0.20, 121°C, sterilization for 20 min.

[0083] L-MRS medium: Add 0.05% (w / v) L-cysteine hydrochloride to MRS medium, pH = 6.80 ± 0.20, sterilize at 121°C for 20 minutes.

[0084] The activation of bacterial strains and preparation of bacterial suspensions in the following experimental examples are as follows:

[0085] Activation culture of Bifidobacterium breve: The strain was removed from a -80°C freezer and inoculated into LMRS anaerobic tube liquid culture medium at a 2% (v / v) inoculation rate. The culture was incubated at 37°C to obtain the first-generation Bifidobacterium breve. The strain was then inoculated into LMRS anaerobic tube liquid culture medium at a 2% (v / v) inoculation rate. The culture was incubated at 37°C for 16 h to obtain the second-generation Bifidobacterium breve BBr26 bacterial solution for use.

[0086] Preparation of Bifidobacterium breve BBr26 bacterial suspension: Bifidobacterium breve BBr26 was activated for 2 generations according to a 2% inoculum size. The cultured Bifidobacterium breve bacterial suspension was centrifuged at 10,000 rpm for 1 min. The supernatant was poured out and the cells were washed twice with an equal volume of PBS and resuspended in the same volume of PBS to obtain a Bifidobacterium breve BBr26 bacterial suspension for use.

[0087] Experimental Example 1: Isolation and identification of Bifidobacterium breve

[0088] (1) Isolation and screening of Bifidobacterium breve

[0089] The collected infant feces were placed in a sterile sampling tube and transported in an ice box. They were diluted with 0.85% saline gradients under sterile conditions, and the appropriate dilution gradient was selected and spread on LMRS + lithium salt agar plates and cultured at 37°C for 48-72 hours. The suspected single colonies were picked by observing the colony morphology with the naked eye, and they were observed under a microscope and preliminarily screened and purified. After purification, the LMRS liquid anaerobic tube was used to culture at 37°C for 8-12 hours, the supernatant was centrifuged and resuspended in a sterile 25% glycerol aqueous solution and stored in the strain bank of Wuhan Weikang Probiotics Research Institute.

[0090] (2) Identification of Bifidobacterium breve:

[0091] The target strains were cultured in liquid, the bacteria were collected, and the genomic DNA was extracted. The universal primers 27F and 1492R described in paragraph 58 of Chinese invention patent ZL202210478937.4 were used to amplify the 16SrDNA fragment. The PCR amplification products were detected by agarose gel electrophoresis, and the PCR products were sequenced. The PCR reaction system included: 10× buffer 10μL, 10mM dNTP 2μL, 1μL of upper and lower primers, 2μL of DNA template, 0.5μL of Taq enzyme, and 34μL of ddH2O. Pre-denaturation at 95℃ for 10min; then 35 cycles of 94℃ for 30s, 60℃ for 30s, and 72℃ for 1min, followed by extension at 72℃ for 5min. The PCR products were detected by gel electrophoresis and sent to Wuhan Jinkairui Bioengineering Co., Ltd. for sequencing. The identified gene sequences were compared in the NCBI database using the BLAST tool. According to the results of molecular biological identification, the Latin name of the strain category is Bifidobacterium breve , and the strain was identified as Bifidobacterium breve.

[0092] Experimental Example 2: Screening of Bifidobacterium breve for its ability to inhibit hyaluronidase activity

[0093] (1) Activate the bacteria to the second generation. Add 0.1 mL of 2.5 mmol / L CaCl2 and 0.5 mL of 600 U / mL hyaluronidase to the test tube and treat at 37℃ for 20 min; add 0.5 mL of the test bacterial suspension and treat at 37℃ for 20 min; add 0.5 mL of 0.5 mg / mL sodium hyaluronate solution and treat at 37℃ for 30 min; let it stand at room temperature for 5 min, add 0.5 mL of acetylacetone solution (1.4 mL of acetylacetone dissolved in 20 mL of 1.0 mol / L Na2CO3 solution, prepared immediately) and 0.1 mL of 0.4 mol / L NaOH solution, boil in water bath for 15 min, and immediately ice bath for 5 min; add 1 mL of Ehrlich reagent (weigh 0.8 g of p-dimethylaminobenzaldehyde dissolved in 15 mL of concentrated hydrochloric acid and 15 mL of anhydrous ethanol, prepared immediately), color at room temperature for 20 min, and incubate at 530℃ for 30 min. The absorbance was measured at a wavelength of 100 nm, with M-16V as the control strain.

[0094] (2) Calculation method: Inhibition rate / % = [(AB)-(CD)] / (AB) Where: A is the absorbance of the control group (acetic acid buffer replaces bacterial suspension); B is the absorbance of the blank group (acetic acid buffer replaces enzyme solution and bacterial suspension); C is the absorbance of the experimental group; D is the absorbance of the blank group

[0095] Table 1. Results of inhibition of hyaluronidase activity by Bifidobacterium breve

[0096]

[0097] Hyaluronidase is a hydrolase that degrades hyaluronic acid and is a major component of the extracellular matrix of connective tissue. It is present in most IgE-mediated type I and T cell-mediated type IV allergic reactions. The vast majority of allergic diseases are type I allergic reactions associated with hyaluronidase activity in vivo. Therefore, the in vitro hyaluronidase inhibition assay is an effective method for rapid screening of anti-allergic substances. Five strains of Bifidobacterium breve isolated from infant feces were screened for hyaluronidase inhibition activity. Bifidobacterium breve M-16V, which has been reported to have anti-allergic effects, was used as a control. The results are shown in Table 1. Two strains of Bifidobacterium breve were identified that showed stronger hyaluronidase inhibition than M-16V. One strain, Bifidobacterium breve 4, exhibited significantly greater inhibition of hyaluronic acid activity than M-16V. This suggests that Bifidobacterium breve 4 has preliminary potential to inhibit hyaluronidase activity, correcting Th1 / Th2 imbalance and alleviating allergic symptoms. This may alleviate IgE-mediated allergic reactions. The strain was named Bifidobacterium breve BBr26 and deposited. The deposit information is as follows:

[0098] Deposit number: CGMCC NO.32424;

[0099] Classification name: Bifidobacterium breve Bifidobacterium breve ;

[0100] Deposit date: October 30, 2024;

[0101] Depository: China General Microbiological Culture Collection Center;

[0102] Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0103] Experimental Example 3: Bifidobacterium breve BBr26 inhibits tryptase secretion activity of mast cells

[0104] (1) Mast cell culture: Cultivate mast cells: Aspirate the cell suspension in the culture flask with a pipette, centrifuge at 800 rpm / min for 3 minutes, discard the supernatant, resuspend with an equal volume of culture medium, and subculture at a ratio of 1:3.

[0105] (2) Preparation of Bifidobacterium breve BBr26: Activate the strain for three generations. Collect the cells by centrifugation in 1 ml of the bacterial suspension, wash twice with sterile PBS, and resuspend in DMEM medium. Finally, dilute the bacterial suspension to the desired cell concentration based on the cell count and set aside.

[0106] (3) The mastoma cells cultured for 24 h were taken out of the constant temperature incubator and placed in a clean bench. The experiment set up three treatments, namely the probiotic treatment group (BBr26): 0.5 ml of Bifidobacterium brevis BBr26 suspension was inoculated into each well to make the bacterial suspension concentration reach 2 × 10 8 CFU / mL; positive drug control group (C48 / 80): add 0.5 ml of DMEM medium; blank control group (CK): add 0.5 ml of DMEM medium. The above 24-well plate was placed in a 37°C constant temperature incubator and incubated for 3 hours. After the incubation period, 100 μl of 100 μg / ml C48 / 80 was added to the probiotic treatment group and the positive drug control group, and 100 μl of DMEM medium was added to the blank control group, and the plates were incubated at 37°C for 1 hour. After the incubation period, the reaction was terminated by placing the plates on ice for 10 minutes. The 24-well plate was placed in a clean bench, and all the liquid in the wells was carefully transferred to a sterile centrifuge tube. The plates were centrifuged at 1000 g for 20 minutes. Then, 50 μl of the supernatant was taken and the tryptase concentration was determined using the corresponding ELISA kit.

[0107] Table 2. Inhibition of tryptase secretion by mast cells by Bifidobacterium breve BBr26

[0108]

[0109] When allergies cause mast cell degranulation, a large amount of tryptase enters the bloodstream, causing a sharp increase in serum concentration. Serum tryptase has been widely used in clinical practice to diagnose mast cell-related diseases, including allergic reactions, mastocytosis, and many other secondary or idiopathic mast cell activation syndromes. The experimental results are shown in Table 2. Figure 1 As shown, the tryptase content secreted by mast cells in the Bifidobacterium breve BBr26 group was 217.15 ng / mL, which was significantly lower than that in the blank control group (CK) and the positive drug group (C48 / 80), indicating that Bifidobacterium breve BBr26 has the effect of alleviating allergic reactions and inhibiting many other secondary or idiopathic mast cell activation syndromes such as mastocytosis.

[0110] Experimental Example 4: Bifidobacterium breve BBr26 promotes INF-γ secretion in mouse spleen cells

[0111] (1) Preparation of Bifidobacterium breve BBr26 bacterial suspension: Activate the strain for three generations. Collect the cells by centrifugation in 1 ml of the bacterial suspension, then wash twice with sterile PBS and resuspend in DMEM medium. Finally, dilute the bacterial suspension to the desired cell concentration based on the cell count and set aside.

[0112] (2) Balb / c mice were killed by cervical dislocation, and the spleen was removed and placed in pre-cooled RPMI-1640 medium containing 5% FBS; the spleen was ground with the smooth end of a syringe in a 70 μm mesh until there was no obvious tissue block, and the cells were filtered into a 50 ml centrifuge tube, centrifuged at 500 g for 5 minutes, and the supernatant was discarded; the cells at the bottom of the centrifuge tube were dispersed, and after precipitation, 2-5 ml of red blood cell lysis buffer was added. After about 5 minutes, phosphate buffer solution (PBS) was added to terminate the reaction and centrifuged; a certain volume (about 5 ml) of complete cell culture medium (RPMI-1640 + 10% FBS + 1% penicillin and streptomycin + 50 μM β-mercaptoethanol) was added and mixed to obtain a single-cell suspension; the single-cell suspension was stained with trypan blue, counted under a microscope, and placed on ice for later use.

[0113] (3) The spleen cells were cultured at 4×10 6 cells / mL was added to a 96-well plate (100 μL per well), and then the Bifidobacterium breve suspension was added at a concentration of 4×10 6 CFU / well (i.e. 2×10 7 CFU / mL) were added to a 96-well plate (100 μL per well). 100 μL of culture medium served as the blank control (NC), 100 μL of 2 μg / mL lipopolysaccharide (LPS) served as the positive control-1, and 100 μL of 4 μg / mL concanavalin A (conA) served as the positive control-2. The treated spleen cells were then cultured for 48 hours, the supernatant was centrifuged, and analyzed using commercially available IFN-γ and IL-12 ELISA kits.

[0114] Table 3. Bifidobacterium breve BBr26 promotes the secretion of interferon-γ in mouse spleen cells

[0115]

[0116] Type I hypersensitivity reactions mediated by specific IgE antibodies are primarily manifested by an imbalance in the Th1 / Th2 immune system within the allergic body. Th2 cells secrete excessive amounts of cytokines such as interleukin-4 (IL-4), stimulating B cells to produce excessive amounts of IgE antibodies. Interferon-γ (IFN-γ) can inhibit the overproduction of cytokines in Th2 lymphocytes, particularly IL-4, thereby reducing B cell proliferation. IFN-γ can also stimulate Th1 immune responses and inhibit IgE production. The experimental results, shown in Table 3, show that both the positive control LPS group and the CoaA group significantly increased INF-γ levels, indicating that the mouse spleen cells extracted from this study can secrete INF-γ normally. Compared with the blank control group, Bifidobacterium breve BBr26 enhanced INF-γ secretion in mouse spleen cells. This suggests that Bifidobacterium breve BBr26 has the potential to alleviate type I hypersensitivity reactions mediated by specific IgE antibodies.

[0117] Bifidobacterium breve BBr26 was evaluated by inhibiting hyaluronidase activity, inhibiting mast cell tryptase secretion activity, and promoting mouse spleen cells, indicating that Bifidobacterium breve BBr26 can alleviate polytype hypersensitivity reactions.

[0118] Experimental Example 5: Tolerance of Bifidobacterium breve BBr26 to Simulated Gastric Fluid

[0119] (1) Preparation of reagents: Preparation of artificial gastric juice: Prepare 0.5% normal saline, add 0.3% pepsin, adjust the pH to 3 with dilute hydrochloric acid 1M HCl, fully dissolve, filter and sterilize with a 0.22μm microporous filter membrane;

[0120] (2) Preparation of experimental strains: According to the preparation method of Bifidobacterium breve BBr26 bacterial suspension, obtain Bifidobacterium breve BBr26 bacterial suspension, and adjust the number of live bacteria in the bacterial suspension to 10 according to the results of the previous live bacteria count. 8 The experiment was performed with 1000 CFU / mL.

[0121] (3) Artificial gastric juice resistance test: 0.1 mL of bacterial suspension was added to 0.9 mL of artificial gastric juice (pH = 3) and incubated at 37 °C for 3 h. The treated solution was gradient diluted at 0 h and 3 h, and then spread on LMRS plates for viable bacterial count. The survival rate of the strain in artificial gastric juice and bile salt was calculated according to the following calculation formula:

[0122]

[0123] Wherein, N1 represents the number of viable bacteria in the strain system after treatment, 1g (CFU / mL); N0 represents the initial number of viable bacteria in the strain system, that is, the number of viable bacteria measured at 0h, 1g (CFU / mL)

[0124] Table 4. Tolerance results of Bifidobacterium breve in artificial gastric fluid (pH=3)

[0125]

[0126] According to the experimental results in Table 4, the survival rate of Bifidobacterium breve BBr26 after being treated in artificial gastric juice (pH=3) for 3 hours was 72.42%, indicating that it has good tolerance and that Bifidobacterium breve BBr26 can survive in a low-acid gastric juice environment.

[0127] Experimental Example 6: Bifidobacterium breve BBr26 inhibits the growth of pathogenic bacteria

[0128] Preparation of Escherichia coli ATCC25922 suspension: Escherichia coli ATCC25922 was inoculated into LB liquid medium at 2% (v / v), cultured at 37°C for 16-20 hours, and then the bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0129] Preparation of Salmonella Enteritidis ATCC14028 suspension: Salmonella Enteritidis ATCC14028 was inoculated into LB liquid medium at 2% (v / v), cultured at 37°C for 16-20 hours, and then the bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0130] Preparation of Staphylococcus aureus ATCC25923 suspension: Staphylococcus aureus ATCC25923 was inoculated into LB liquid medium at 2% (v / v), cultured at 37°C for 16-20 h, and then the bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0131] Preparation of Streptococcus Parisi suspension: Bacteroides fragilis was inoculated into BHI blood plates containing 5% defibrinated sheep blood, cultured at 37°C for 16-20 hours, and the bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0132] Preparation of Streptococcus pasteurianus suspension: Streptococcus pasteurianus was inoculated into a BHI blood plate containing 5% defibrinated sheep blood and cultured at 37°C for 16-20 hours. The concentration of the bacterial suspension was adjusted to 10 8 CFU / mL.

[0133] Preparation of Streptococcus anginosus MW090399 bacterial suspension: Streptococcus anginosus was inoculated into TSA blood plates containing 5% defibrinated sheep blood and cultured at 37°C for 16-20 hours. The bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0134] Preparation of saprophytic Staphylococcus suspension: Saprophytic Staphylococcus was inoculated into LB liquid medium at 2% (v / v), cultured at 37°C for 16-20 h, and then the concentration of the suspension was adjusted to 10 8CFU / mL.

[0135] Preparation of Propionibacterium acnes suspension: Propionibacterium acnes was inoculated into TSA blood plates containing 5% defibrinated sheep blood and cultured at 37°C for 48 h. The bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0136] Bacteriostasis test: Cool the MRS broth medium containing 1.5% agar (BHI medium containing 5% defibrinated sheep blood for Streptococcus Parisi, Streptococcus pasteurianus, and Streptococcus anginosus; TSA medium containing 5% defibrinated sheep blood for Propionibacterium acnes) to about 55°C and mix it with the indicator bacteria suspension in a certain proportion so that the number of viable indicator bacteria is 10 6 CFU / mL, and then quickly pour it into the plate with the Oxford cup placed in advance. After the culture medium cools and solidifies, take out the Oxford cup and inject 200μL of the strain fermentation liquid into each well (the number of viable bacteria is 10 8 CFU level), and the diameter of the inhibition zone was measured after overnight incubation at 37°C.

[0137] Table 5. Inhibition of pathogen growth by Bifidobacterium breve BBr26

[0138]

[0139] As shown in Table 5, Bifidobacterium breve BBr26 has a significant inhibitory effect on Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, and Propionibacterium acnes. Staphylococcus aureus and Propionibacterium acnes are common pathogens on the surface of the skin, indicating that Bifidobacterium breve BBr26 can effectively inhibit the growth of common pathogens on the surface of the skin.

[0140] Experimental Example 7: Study on the proliferation of Bifidobacterium breve BBr26 on macrophage RAW264.7

[0141] Activation of Bifidobacterium breve BBr26: Remove strain BBr26 from a -80°C freezer, inoculate at a 2% (v / v) inoculum into LMRS anaerobic tube liquid culture medium, and incubate at 37°C to obtain a first-generation B. breve BBr26 bacterial suspension. Inoculate again at a 2% (v / v) inoculum into LMRS anaerobic tube liquid culture medium, and incubate at 37°C for 16 h to obtain a second-generation B. breve BBr26 bacterial suspension for later use.

[0142] RAW264.7 cell culture: After thawing, remove RAW264.7 cells from the freezer and resuspend in complete DMEM medium supplemented with 1% nonessential amino acids, 1% penicillin-streptomycin, and 10% fetal bovine serum (FBS). Culture in a 5% CO2 incubator at 37°C. Change the medium every other day. Subculture when the cells reach a confluency of 80% or higher. Wash twice with sterile PBS, add 2 mL of culture medium, gently scrape the cells with a cell scraper, centrifuge, discard the supernatant, and resuspend in complete DMEM medium. Subculture at a 1:3 ratio.

[0143] The concentration of RAW264.7 cells was adjusted to 1 × 10 4 Cells were seeded into 96-well plates at a concentration of 100 μL / well at 100 cells / mL and cultured for 24 hours. The supernatant was then aspirated and the plates were washed twice with PBS. The experiment was divided into a blank group and a BBr26 group. In the blank group, only 100 μL of DMEM medium was added to the wells and the plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. In the BBr26 group, 100 μL of a bacterial suspension (a viable bacterial suspension was washed twice with PBS, resuspended in DMEM, and diluted to a ratio of 100:1) was added to the wells and cultured for 24 hours. After the incubation period, the supernatant was aspirated, the plates were washed once with PBS, and 100 μL of DMEM medium containing 10% CCK-8 was added. The plates were incubated at 37°C in the dark for 1 hour. The absorbance (OD) of each well was measured at 450 nm using a microplate reader.

[0144] The cell viability was calculated as CV = OD1 / OD2 × 100%.

[0145] Note: CV-cell viability (%); OD1: OD value of each treatment group at 450 nm wavelength; OD2: OD value of the blank group at 450 nm wavelength.

[0146] Table 6. Results of the RAW264.7 cell proliferation assay using Bifidobacterium breve BBr26

[0147]

[0148] Probiotics can enhance the activity of monocytes and macrophages, increasing the levels of reactive oxygen species and reactive nitrogen species, and their phagocytic capacity. They can also process and present macrophage antigens to T and B lymphocytes, triggering humoral and cellular immunity, thereby regulating the body's nonspecific and specific immune functions. Experiments on the proliferation of RAW264.7 macrophages by Bifidobacterium breve BBr26 showed a proliferation rate of 178.32%, significantly higher than that of the commonly used immunomodulatory strain Lactobacillus rhamnosus LGG, indicating that Bifidobacterium breve BBr26 has immunomodulatory properties.

[0149] Experimental Example 8: Verification of the Effect of Bifidobacterium breve BBr26 in Alleviating Atopic Dermatitis in Mice

[0150] 1. Atopic dermatitis model in mice

[0151] Forty healthy, 6-week-old Balb / c mice were randomly divided into four groups of 10 mice each: a blank control group (Control), a model group (DNFB) administered with 2,4-dinitrofluorobenzene (DNFB), a positive drug group (Keto) administered with ketotifen, and an intervention group (Bifidobacterium breve BBr26). The experiment lasted three weeks: the first week was an acclimatization period for the mice; gavage administration began in the second week and continued until the end of the experiment.

[0152] (1) Bifidobacterium breve BBr26 intervention group: Oral administration of Bifidobacterium breve BBr26 bacterial solution, with 0.2 mL of bacterial solution (1×10 10 CFU / mouse / time) by gavage;

[0153] (2) Positive drug intervention group: Oral gavage of ketotifen (Keto) solution, with a dose of 0.2 mL of solution (the total amount of drug for a single oral gavage is 1 mg / kg based on the mouse's body weight) per mouse per time;

[0154] (3) The blank control group (NC) and the model group (MC) were not given Bifidobacterium breve BBr26 bacterial solution or ketotifen (Keto) by gavage, and only an equal amount of normal saline was given by gavage as a control; the second to third weeks were the modeling period. On the first day of modeling, 100 μL of 0.5% DNFB solution was applied to the backs of the mice in the model group, BBr26 intervention group, and positive drug group, and 20 μL of 0.5% DNFB solution was applied to the right ears of the mice in the model group, BBr26 intervention group, and positive drug group for skin sensitization and stimulation. Starting from the fifth day, 100 μL of 0.2% DNFB solution was applied to the backs of the mice in the model group, BBr26 intervention group, and positive drug group, and 20 μL of 0.5% DNFB solution was applied to the right ears of the mice in the model group, BBr26 intervention group, and positive drug group for skin sensitization and stimulation every three days, for a total of 4 times. The blank group of mice received only an equal amount of acetone / olive oil matrix solution applied to their right ears as a control. All groups had free access to water and food. Before modeling, the ear thickness of the six groups of mice was measured using a vernier caliper. After modeling, blood was drawn and the mice were sacrificed. Serum was collected and assayed for relevant inflammatory markers. The spleen and thymus of the mice were collected and the relevant organ indices were measured. The ear thickness of the mice was measured, and the back skin tissue of the mice was extracted for subsequent measurement of relevant indicators.

[0155] Effect of Bifidobacterium breve BBr26 on ear thickness in mice with atopic dermatitis

[0156] The thickness of the right ear of each group of mice was measured with a vernier caliper on days 0, 2, 5, 8, 10, 13, and 15 after modeling. Figure 2 As shown, ear thickness remained essentially unchanged in the blank control group over time. Ear thickness in the model group (DNFB), BBr26-treated group, and positive drug group (Keto) was significantly (p<0.01) higher than in the blank control group, demonstrating the success of the atopic dermatitis mouse model. The right ear thickness of mice in the BBr2-treated group was significantly (p<0.01) lower than that in the model group and positive drug group treated with 0.5% DNFB solution, indicating that Bifidobacterium breve BBr26 can alleviate allergic symptoms in atopic dermatitis mice.

[0157] Effects of Bifidobacterium breve BBr26 on inflammatory infiltration in the skin tissue of mice with atopic dermatitis

[0158] After the experiment, the exposed skin tissues from the backs of mice in each group were extracted and stored in tissue freezing solution, and then H&E stained sections were prepared. Figure 3 As shown, compared with the blank group (Control), the back skin thickness and inflammatory infiltration of the model group (DNFB) mice were significantly increased; Bifidobacterium breve BBr26 significantly reduced the increase in mouse skin thickness and alleviated the skin inflammatory infiltration; it can be seen that Bifidobacterium breve BBr26 can significantly improve skin inflammation in mice with atopic dermatitis.

[0159] 4. Mast Cell Staining in Atopic Dermatitis Mice

[0160] 5µm thick sections were baked, dehydrated with alcohol, and stained with mast cell staining solution. After differentiation, the slides were transparently mounted and prepared for pathological sections. The number of mast cells was counted in three different fields under an optical microscope. After modeling, blood was collected and the mice were sacrificed. Histopathological analysis of the skin from the depilated back of the mice was performed. Histopathological sections of the skin from the back of the mice were stained with toluidine blue. As shown in Figure 4, mast cell infiltration in the skin tissue was reduced. At the same time, the number of mast cells produced was reduced, and the inhibition of skin Bifidobacterium breve BBr26 significantly suppressed tissue inflammation.

[0161] 5. Determination of thymus index and spleen index in atopic dermatitis mice

[0162] After the modeling was completed, blood was collected and the mice were killed. The thymus index and spleen index of the mice were measured. The experimental results are as follows Figure 5 、 Figure 6Compared with the blank control group, the thymus index of the model group decreased significantly, and the spleen index increased significantly. Through the intervention of Bifidobacterium breve BBr26, the abnormal thymus index and spleen index of mice were alleviated. It appears that Bifidobacterium breve BBr26 can alleviate the excessive immune response of allergic mice.

[0163] 6. Determination of inflammatory factors in the serum of mice with atopic dermatitis

[0164] After modeling, blood was collected and the mice were sacrificed. Eye blood was collected and centrifuged at 3000 rpm / min for 15 minutes. The supernatant serum was collected and assayed for IL-4, IL-13, IL-10, and INF-γ levels using enzyme-linked immunosorbent assay kits. The core mechanism of atopic dermatitis is an immune abnormality dominated by a Th2 response. After allergens invade the skin barrier, they are taken up by antigen-presenting cells (DCs) and LCs. DCs then present the allergens to Th0 cells. Under the influence of the cytokine IL-4, Th0 cells differentiate into Th2 cells and proliferate. The Th2 cell-specific nuclear transcription factor GATA3 plays a crucial role in this process. A large number of Th2 cells secrete the cytokines IL-4, IL-5, and IL-13, resulting in an excessive Th2 immune response. The overproduction of these Th2 cytokines leads to a decrease in the expression of loricrin and involucrin, proteins that maintain skin barrier integrity, thereby exacerbating skin barrier damage.

[0165] IL-4 and IL-13 play an important role in the entire Th2 immune response. The results of the measurement of IL-4 and IL-13 in the serum of atopic mice showed that Figure 8 , Bifidobacterium breve BBr26 significantly inhibited the increase of IL-4 and IL-13 in serum, thereby maintaining the integrity of the skin barrier.

[0166] IL-10 helps to inhibit Th2 immune response and restore the Th1 / Th2 immune balance. The results of the IL-10 assay in the serum of atopic mice are as follows: Figure 7 , Bifidobacterium breve BBr26 significantly increased the content of IL-10 in serum, thereby inhibiting the Th2 immune response, restoring the Th1 / Th2 immune balance, and alleviating the Th2 immune response.

[0167] The expression of IFN-γ associated with Th1 immune response can inhibit IgE synthesis, Th2 cell differentiation, and IL-4 receptor expression. The results of the measurement of IL-10 in the serum of atopic mice are as follows: Figure 8 , Bifidobacterium breve BBr26 significantly increased the content of IFN-γ in serum, thereby inhibiting IgE synthesis, Th2 cell differentiation, and IL-4 receptor expression.

[0168] VII. Determination of IgE Content in Atopic Dermatitis Mice

[0169] The core immune mechanism of atopic dermatitis is an excessive Th2 immune response. Th2 immune cells induce the synthesis of IgE and the infiltration of inflammatory cells by releasing cytokines such as IL-4, IL-5, and IL-13. The results of IgE determination in serum samples of atopic dermatitis mice are as follows: Figure 8 , Bifidobacterium breve BBr26 significantly reduced the synthesis of IgE in mice and alleviated the excessive Th2 immune response.

Claims

1. A Bifidobacterium breve that inhibits hyaluronidase activity and relieves allergies ( Bifidobacterium breve )BBr26 ， It is characterized in that its preservation number is CGMCC NO.32424.

2. Bifidobacterium breve with a deposit number of CGMCC NO.32424 ( Bifidobacterium breve ) Use of BBr26 in preparing a medicine for relieving atopic dermatitis, and / or a food for enhancing immune function, and / or an antibacterial agent, characterized in that: The immune enhancement includes: increasing the proliferation rate of immune cells RAW264.7, promoting the secretion of INF-γ by spleen cells, inhibiting the secretion of tryptase by mast cells, and regulating cytokine levels; The antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus pasteurianus, Staphylococcus saprophyticus, and Propionibacterium acnes.

3. according to claim 2, the deposit number is CGMCC NO.32424 Bifidobacterium breve ( Bifidobacterium breve ) Use of BBr26 in preparing a medicine for relieving atopic dermatitis, and / or a food for enhancing immune function, and / or an antibacterial agent, characterized in that: The atopic dermatitis relief includes: reducing ear thickness, reducing skin thickness, relieving skin inflammatory infiltration, reducing mast cell count, increasing thymus index, reducing spleen index, inhibiting hyaluronidase activity, and regulating cytokine levels.

4. according to claim 3, the deposit number is CGMCC NO.32424 Bifidobacterium breve ( Bifidobacterium breve ) Use of BBr26 in preparing a medicine for relieving atopic dermatitis, and / or a food for enhancing immune function, and / or an antibacterial agent, characterized in that: The regulating cytokine levels includes: reducing the levels of IL-4 and IL-13, and increasing the levels of IL-10 and INF-γ.

5. A drug for relieving atopic dermatitis, comprising: Active ingredient; characterized in that the active ingredient includes: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

6. A drug for alleviating atopic dermatitis according to claim 4, characterized in that: Also includes: Auxiliary materials.

7. A food for enhancing immune function, comprising: Active ingredient; characterized in that the active ingredient includes: Bifidobacterium breve ( Bifidobacterium breve ) BBr26; the immune enhancement includes: increasing the proliferation rate of immune cells RAW264.7, promoting spleen cells to secrete INF-γ, inhibiting mast cells to secrete tryptase, and regulating cytokine levels.

8. The food for enhancing immune function according to claim 7, characterized in that: Also includes: Auxiliary materials.

9. A bacteriostatic agent comprising: Active ingredient; characterized in that the active ingredient includes: Bifidobacterium breve ( Bifidobacterium breve ) BBr26; The antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus pasteurianus, Staphylococcus saprophyticus, and Propionibacterium acnes.

10. The antibacterial agent according to claim 9, characterized in that Also includes: Auxiliary materials.

11. An in vitro antibacterial method, characterized in that: The collection number is CGMCC NO.32424. Bifidobacterium breve ) BBr26 is antibacterial.

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