Bifidobacterium breve BBr26 capable of inhibiting hyaluronidase activity and relieving allergy and application, product and method thereof

Abstract

The invention discloses bifidobacterium breve BBr26 capable of inhibiting hyaluronidase activity and relieving allergy as well as an application, a product and a method of the bifidobacterium breve BBr26, and belongs to the technical field of microorganisms. The invention provides a Bifidobacterium breve (Bifidobacterium breve) strain BBr26 with anti-allergy and / or immunoregulation capability, and the preservation number of the Bifidobacterium breve strain BBr26 is CGMCC (China General Microbiological Culture Collection Center) NO.32424. The invention also provides a preparation method of the Bifidobacterium breve strain The invention also provides an application of the strain bifidobacterium breve BBr26 in preparation of products for relieving atopic dermatitis, and / or food for enhancing immune function, and / or bacteriostatic agents, and a product and a method based on the strain BBr26. The strain disclosed by the invention has broad-spectrum bacteriostasis, can relieve allergy and atopic dermatitis, and can effectively play a role in immunoregulation.

Description

Technical Field

[0001] The present invention belongs to the technical field of microorganisms, and specifically relates to Bifidobacterium breve BBr26 capable of inhibiting hyaluronidase activity and relieving allergies, and applications, products and methods thereof. Background Art

[0002] The prevalence of allergic diseases has gradually increased in recent years. Common allergic diseases include allergic rhinitis, allergic asthma, allergic eczema, food allergy, atopic dermatitis and contact dermatitis. In allergic diseases, inflammation often occurs. Allergic reactions can be divided into types I, II, III and IV according to their mechanisms. Different types of allergic reactions have different mechanisms. In type I allergic reactions, IgE antibodies bind to IgE receptors on the surface of mast cells and basophils after contact with specific antigens, causing rapid degranulation and release of pharmacological mediators of local inflammation through bridging, resulting in clinical symptoms; in type II allergic reactions, antibodies bind to cell surface antigens themselves or to antigens and haptens adsorbed on the cell surface, and complement participates, resulting in cell and tissue damage; in type III allergic reactions, complement activation and neutrophil release of lytic enzymes cause tissue damage, inhaled antigens cause lung diseases and cause local damage, and systemic immune complexes formed by antibodies and microorganisms or self-antigens cause immune complexes to deposit in blood vessels, leading to 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, causing inflammation or delayed hypersensitivity reactions.

[0003] Hyaluronidase is a hydrolase that degrades hyaluronic acid. It is the main component of the extracellular matrix of the body's connective tissue and is present in most IgE-mediated type I and T-cell-mediated type IV allergic reactions. Most allergic diseases are type I allergic reactions related to hyaluronidase activity in the body. Therefore, the hyaluronidase in vitro inhibition test is an effective method for rapid in vitro screening of anti-allergic substances. At the same time, inhibiting hyaluronidase activity can ensure the normal content and function of hyaluronic acid. Hyaluronidase is used as a drug diffusion agent in clinical practice, especially in tumor treatment and subcutaneous administration. It has a good application prospect.

[0004] Bifidobacterium breve BBr26 is a Gram-positive anaerobic bacterium that is mostly found in infant feces and belongs to the subspecies of Bifidobacterium breve. No reports have been seen on Bifidobacterium breve that can relieve allergic symptoms based on the inhibition of hyaluronidase.

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

[0006] In order to solve the above-mentioned needs in the art, the present invention provides a product and method of Bifidobacterium breve BBr26 for relieving atopic dermatitis.

[0007] The purpose of the present invention is achieved through the following technical solutions: A Bifidobacterium breve (Bifidobacterium breve) that inhibits hyaluronidase activity and relieves allergies Bifidobacterium breve )BBr26, its deposit number is CGMCC NO.32424.

[0008] Bifidobacterium breve (CGMCC NO.32424) Bifidobacterium breve ) BBr26, use in the preparation of products for alleviating atopic dermatitis, and / or foods for enhancing immune function, and / or antibacterial agents.

[0009] 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; Preferably, the immunity 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; Preferably, the regulating cytokine levels comprises: reducing the levels of IL-4 and IL-13, and increasing the levels of IL-10 and INF-γ; Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, Propionibacterium acnes.

[0010] A product for relieving atopic dermatitis, comprising: an active ingredient; the active ingredient comprises: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0011] The product for alleviating atopic dermatitis further comprises: auxiliary materials.

[0012] A food for enhancing immune function, comprising: an active ingredient; the active ingredient comprises: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0013] The food for enhancing immune function also includes auxiliary materials.

[0014] An antibacterial agent, comprising: an active ingredient; the active ingredient comprises: Bifidobacterium breve (Bifidobacterium breve )BBr26.

[0015] The antibacterial agent further comprises: auxiliary materials.

[0016] An in vitro antibacterial method using Bifidobacterium breve (CGMCC NO.32424) Bifidobacterium breve )BBr26 is antibacterial.

[0017] The beneficial effects of the present invention are as follows: The present invention screened a new strain from infant feces samples, and molecular identification showed that the strain was Bifidobacterium breve ( Bifidobacterium breve ), and named it strain BBr26. The present invention has been confirmed by 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 hyaluronidase activity, inhibit mast cell secretion of tryptase, promote mouse spleen cells to secrete INF-γ, and relieve mouse atopic dermatitis. At the same time, the strain BBr26 also has a significant effect on the proliferation rate of macrophages RAW264.7, and effectively regulates the proliferation of macrophages RAW264.7 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, saprophytic Staphylococcus, and Propionibacterium acnes.

[0018] The Bifidobacterium breve of the present invention ( Bifidobacterium breve )The deposit information of BBr26 is as follows: Deposit number: CGMCC NO.32424; Classification name: Bifidobacterium breve Bifidobacterium breve ; Deposit date: October 30, 2024; Depository: General Microbiology Center, China Microbiological Culture Collection Administration; Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a bar graph showing the results of measuring the activity of Bifidobacterium breve BBr26 in inhibiting mast cell secretion of trypsin in Experimental Example 3 of the present invention.

[0020] Figure 2 It is a line graph of the ear thickness changes of each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0021] Figure 3These are skin tissue sections of each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0022] Figure 4 This is a microscopic examination of mast cell staining in atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0023] Figure 5 It is a bar graph of the thymus index of each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0024] Figure 6 It is a bar graph of the spleen index of each group of atopic dermatitis model mice in Experimental Example 8 of the present invention.

[0025] Figure 7 This is a bar graph of the levels of serum IL-4, IL-13, IL-10, and INF-γ in each group of mice in the atopic dermatitis model of Experimental Example 8 of the present invention.

[0026] Figure 8 This is a bar graph of the serum IgE content of each group of mice in the atopic dermatitis model of Experimental Example 8 of the present invention.

[0027] Figure 1 The meanings of the marks are listed as follows: CK: blank control group; C48 / 80: positive drug control group; BBr26: probiotic treatment group.

[0028] Figures 2 - 8 The meanings of the marks are listed as follows: NC: blank control group; MC: model group; KT: positive drug intervention group; BBr26: Bifidobacterium breve BBr26 intervention group. DETAILED DESCRIPTION

[0029] The details of the present invention are further described below in conjunction with specific embodiments and experimental examples, but the protection scope of the present invention is not limited thereto.

[0030] Source of biological material 1. Bifidobacterium breve M-16V used in Experimental Example 2 of the present invention is commercially available; 2. Experimental Example 3 of the Present Invention Mast cells were purchased from China Center for Type Culture Collection.

[0031] 3. The Balb / c mice used in Experimental Example 4 of the present invention were purchased from Hubei Safety Evaluation Center.

[0032] 4. The Escherichia coli ATCC25922, Salmonella enteritidis ATCC14028, and Staphylococcus aureus ATCC25923 used in Experimental Example 6 of the present invention were purchased from 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; Streptococcus anginosus was Streptococcus pharyngitis 4997, which has been registered in NCBI with the accession number MT459365; it is currently preserved in the applicant's laboratory; Staphylococcus saprophyticus and Propionibacterium acnes are preserved by Wuhan Weikang Probiotics Research Institute Co., Ltd. The applicant promises to release it to the public within 20 years from the date of application for verification of the technical effects of the present invention.

[0033] 5. The macrophage RAW264.7 used in Experimental Example 7 of the present invention is commercially available.

[0034] VI. The Balb / c mice used in Experimental Example 8 of the present invention were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd.

[0035] The first group of examples, Bifidobacterium breve BBr26 of the present invention This group of embodiments provides a method for inhibiting hyaluronidase activity and relieving allergies. Bifidobacterium breve ) BBr26. All embodiments of this group have the following common features: the Bifidobacterium breve ( Bifidobacterium breve )The deposit number of BBr26 is CGMCC NO.32424.

[0036] Any act of culturing, propagating, fermenting, enriching, producing, preparing, using, inoculating, amplifying, transforming, modifying, transforming, selling, or promising to sell a strain of Bifidobacterium breve BBr26 with a preservation number of CGMCC NO. 32424, and / or using a strain of Bifidobacterium breve BBr26 with a preservation number of CGMCC NO. 32424 in combination with other probiotics, and / or using a strain of Bifidobacterium breve BBr26 with a preservation number of CGMCC NO. 32424 to antagonize various pathogens, and / or prepare an antibacterial agent, and / or prepare an immune-enhancing food, and / or a functional food for regulating intestinal flora falls within the protection scope of the present invention.

[0037] The other probiotics include but are not limited to: Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii lactic acid subsp., Lactobacillus helveticus, Lactobacillus casei, Lactobacillus crispatus, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus salivarius, Saccharomyces cerevisiae, Delb's spore-forming torula, Candida, Wickham's yeast, Pichia, Saccharomyces boulardii, Torulopsis spp., Saccharomyces cerevisiae, Rhodotorula, Schizosaccharomyces pombe , Saccharomyces spp., Bacillus thuringiensis, Bacillus 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.

[0038] Those skilled in the art can make conventional selections or adjustments to pharmaceutical excipients according to actual production needs, combined with 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.), and then make a strain of 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 without technical barriers for those skilled in the art.

[0039] The second group of examples, application of Bifidobacterium breve BBr26 of the present invention This group of embodiments provides Bifidobacterium breve ( Bifidobacterium breve ) BBr26, use in the preparation of products for alleviating atopic dermatitis, and / or foods for enhancing immune function, and / or antibacterial agents.

[0040] 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; Preferably, the immunity 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; Preferably, the regulating cytokine levels comprises: reducing the levels of IL-4 and IL-13, and increasing the levels of IL-10 and INF-γ; Preferably, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, Propionibacterium acnes.

[0041] The third group of embodiments, the product for relieving atopic dermatitis of the present invention This group of embodiments provides a product for relieving atopic dermatitis. All embodiments in this group have the following common features: the product for relieving atopic dermatitis comprises: an active ingredient; the active ingredient comprises: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0042] In a further embodiment, the product for alleviating atopic dermatitis is characterized by further comprising: auxiliary materials.

[0043] In a more specific embodiment, the excipient is 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, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0044] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of preparation preparation (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.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare Bifidobacterium breve BBr26 with a preservation number of CGMCC NO. 32424 into different dosage forms, such as powders, tablets, oral liquids, gels, patches, sprays, lotions, granules, etc.

[0045] In a specific embodiment, the dosage form of the product for relieving atopic dermatitis is selected from: one or more of powder, tablet, liquid, and capsule.

[0046] The fourth group of embodiments, the food for enhancing immune function of the present invention 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 includes: active ingredients; the active ingredients include: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

[0047] In a further embodiment, the immune function enhancing food further includes: auxiliary materials.

[0048] In a more specific embodiment, the excipient is 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, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0049] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of preparation preparation (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.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare Bifidobacterium breve BBr26 with a preservation number of CGMCC NO. 32424 into different dosage forms, such as powders, tablets, oral liquids, gels, patches, sprays, lotions, granules, etc.

[0050] In a specific embodiment, the dosage form of the food for enhancing immune function is selected from: one or more of powder, tablet, liquid, and capsule.

[0051] The fifth group of embodiments, the antibacterial agent of the present invention 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.

[0052] In a further embodiment, the antibacterial agent further comprises: auxiliary materials.

[0053] In a more specific embodiment, the excipient is 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, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants, deflocculating agents, filter aids, release retardants, etc.

[0054] According to the content of the present invention, for different needs in actual production applications, combined with conventional technical means in the field of preparation preparation (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.), those skilled in the art can select and formulate the above-mentioned excipients, and prepare Bifidobacterium breve BBr26 with a preservation number of CGMCC NO. 32424 into different dosage forms, such as powders, tablets, oral liquids, gels, patches, sprays, lotions, granules, etc.

[0055] In a specific embodiment, the dosage form of the antibacterial agent is selected from: one or more of powder, tablet, liquid, and capsule.

[0056] The sixth group of embodiments, the in vitro antibacterial method of the present invention 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.

[0057] In a specific embodiment, the antibacterial spectrum includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, Propionibacterium acnes.

[0058] “Above” and “below” in this article include the original number.

[0059] 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.

[0060] The culture medium and reagents used in the experimental examples of the present invention are as follows: 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 0.05g / L, pH=6.80 ± 0.20, 121℃, sterilization for 20 min.

[0061] L-MRS medium: Add 0.05% (w / v) L-cysteine ​​hydrochloride to MRS medium, pH = 6.80 ± 0.20, 121°C, sterilize for 20 minutes.

[0062] The activation of bacterial strains and the preparation of bacterial suspension in the following experimental examples are as follows: Activation culture of Bifidobacterium breve: the strain was taken out from the -80℃ refrigerator, inoculated in LMRS anaerobic tube liquid culture medium at a rate of 2% (v / v), and cultured at 37℃ to obtain the first-generation Bifidobacterium breve; then inoculated in LMRS anaerobic tube liquid culture medium at a rate of 2% (v / v), and cultured at 37℃ for 16h to obtain the second-generation Bifidobacterium breve BBr26 bacterial liquid for use.

[0063] Preparation of Bifidobacterium breve BBr26 bacterial suspension: Bifidobacterium breve BBr26 was activated for 2 generations according to a 2% inoculation rate. The cultured Bifidobacterium breve bacterial liquid was centrifuged at 10,000 r / min for 1 min, the supernatant was poured out, the bacteria were washed twice with an equal volume of PBS, and then resuspended with the same volume of PBS to obtain Bifidobacterium breve BBr26 bacterial suspension for use.

[0064] Experimental Example 1: Isolation and identification of Bifidobacterium breve (1) Isolation and screening of Bifidobacterium breve The collected infant feces were placed in a sterile sampling tube and transported in an ice box. They were diluted with 0.85% saline gradient under sterile conditions, and the appropriate dilution gradient was selected and spread on the LMRS + lithium salt agar plate, 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, LMRS liquid anaerobic tubes were used for culture at 37°C for 8-12 hours, and 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.

[0065] (2) Identification of Bifidobacterium breve: The target strains were cultured in liquid, the cells were collected, the genomic DNA was extracted, and the universal primers 27F and 1492R recorded 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: 10×buffer 10μL, 10mMdNTP 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 94℃ 30s, 60℃ 30s, and 72℃ 1min for a total of 35 cycles, and then extended 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 , the strain was determined to be Bifidobacterium breve.

[0066] Experimental Example 2: Screening of Bifidobacterium breve with the ability to inhibit hyaluronidase activity (1) Activate the strain to the second generation. Add 0.1 mL of 2.5 mmol / L CaCl2 and 0.5 mL of 600U / 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 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 nm, and M-16V was used as the control strain.

[0067] (2) Calculation method: Inhibition rate / % = [(AB)-(CD)] / (AB) Where: A is the absorbance of the control group (acetic acid buffer replaces the bacterial suspension); B is the absorbance of the blank group (acetic acid buffer replaces the enzyme solution and bacterial suspension); C is the absorbance of the experimental group; D is the absorbance of the blank group Table 1. Results of inhibition of hyaluronidase activity by Bifidobacterium breve

[0068] Hyaluronidase is a hydrolase that degrades hyaluronic acid. It is the main component of the extracellular matrix of the body's connective tissue and is present in most IgE-mediated type I and T-cell-mediated type IV allergic reactions. Most allergic diseases are type I allergic reactions associated with hyaluronidase activity in the body. Therefore, the in vitro inhibition test of hyaluronidase is an effective method for rapid in vitro screening of anti-allergic substances. Five strains of Bifidobacterium breve isolated from infant feces were screened for inhibition of hyaluronidase activity, and Bifidobacterium breve M-16V, which has been reported to have allergy efficacy, was used as a control. The experimental results are shown in Table 1. Two strains of Bifidobacterium breve were screened for inhibition of hyaluronidase activity that was stronger than M-16V, and one strain of Bifidobacterium breve 4 had a significantly stronger ability to inhibit hyaluronic acid activity than Bifidobacterium breve M-16V, indicating that Bifidobacterium breve 4 has the initial ability to inhibit hyaluronidase activity, correct Th1 / Th2 imbalance and relieve allergic symptoms. Relieve IgE-mediated allergic reactions. The strain was named Bifidobacterium breve BBr26 and deposited. The deposit information is as follows: Deposit number: CGMCC NO.32424; Classification name: Bifidobacterium breve Bifidobacterium breve ; Deposit date: October 30, 2024; Depository: General Microbiology Center, China Microbiological Culture Collection Administration; Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0069] Experimental Example 3: Bifidobacterium breve BBr26 inhibits trypsin secretion activity of mast cells (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.

[0070] (2) Preparation of Bifidobacterium breve BBr26: Activate the strain for three generations. Take 1 ml of the bacterial solution and centrifuge to collect the cells, then wash twice with sterile PBS and finally resuspend in DMEM medium. Finally, dilute the bacterial suspension to the required cell concentration according to the cell number and set aside.

[0071] (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 breve BBr26 bacterial suspension was inoculated into each well to make the bacterial suspension concentration reach 2 × 10 8CFU / 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 for 3 hours. After the incubation, 100μl of 100μg / ml C48 / 80 was added to the probiotic treatment group and the positive drug control group, and 100μl of DMEM medium was added to the blank control group, and they were placed at 37°C for 1 hour. After the incubation, ice bath for 10 minutes to terminate the reaction. Place the 24-well plate in an ultra-clean workbench, carefully transfer all the liquid in the well to a sterile centrifuge tube, and centrifuge at 1000g for 20 minutes; then take 50μl of the supernatant and determine the concentration of tryptase using the corresponding ELISA kit; Table 2. Inhibition of tryptase secretion by mast cells by Bifidobacterium breve BBr26

[0072] When allergies cause mast cell degranulation, a large amount of tryptase enters the blood, 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 content of tryptase 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.

[0073] Experimental Example 4: Bifidobacterium breve BBr26 promotes INF-γ secretion in mouse spleen cells (1) Preparation of Bifidobacterium breve BBr26 bacterial suspension: Activate the strain for three generations. Take 1 ml of the bacterial suspension and centrifuge to collect the cells, then wash twice with sterile PBS and finally resuspend in DMEM medium. Finally, dilute the bacterial suspension to the required cell concentration according to the cell number and set aside.

[0074] (2) Balb / c mice were killed by cervical dislocation, and the spleen was removed and placed in pre-cooled RPMI-1640 culture medium containing 5% FBS. The spleen was ground with the smooth end of a syringe in a 70 μm mesh until no obvious tissue blocks were found. The cells were filtered into a 50 ml centrifuge tube and centrifuged at 500 g for 5 min. The supernatant was discarded. The cells at the bottom of the centrifuge tube were dispersed and precipitated. After about 5 min, 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.

[0075] (3) The spleen cells were collected 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) was added to a 96-well plate (100 μL per well). 100 μL of culture medium was used as the blank control group (NC), 100 μL of 2 μg / mL lipopolysaccharide (LPS) was used as the positive control group-1; 100 μL of 4 μg / mL concanavalin A (conA) was used as the positive control group-2; then, the treated spleen cells were cultured for 48 hours, the supernatant was centrifuged, and analyzed with commercially available IFN-γ and IL-12 ELISA reagents.

[0076] Table 3. Bifidobacterium breve BBr26 promotes the secretion of interferon-γ in mouse spleen cells

[0077] Type I hypersensitivity reactions mediated by specific IgE antibodies are mainly manifested as Th1 / Th2 immune imbalance in allergic bodies. Th2 cells secrete excessive cytokines such as interleukin-4 (IL-4), stimulating B cells to produce excessive IgE antibodies. Interferon-γ (IFN-γ) can inhibit the overexpression of cytokines in Th2 lymphocytes, especially inhibit the secretion of IL-4, thereby reducing the proliferation of B cells. IFN-γ can also stimulate the immune response of Th1 and inhibit the synthesis of IgE. The experimental results are shown in Table 3. The positive control LPS group and the CoaA group can significantly increase the content of INF-γ, indicating that the mouse spleen cells extracted this time can secrete INF-γ normally; compared with the blank control group, Bifidobacterium breve BBr26 can promote the secretion of INF-γ by mouse spleen cells. It shows that Bifidobacterium breve BBr26 has the effect of alleviating type I hypersensitivity reactions mediated by specific IgE antibodies.

[0078] 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 polymorphic hypersensitivity reactions.

[0079] Experimental Example 5: Tolerance of Bifidobacterium breve BBr26 to Simulated Gastric Fluid (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, and filter with a 0.22μm microporous filter membrane to sterilize after fully dissolving; (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 based on the results of the previous live bacteria count. 8 The experiment was performed with 1000 μg of CFU / mL.

[0080] (3) Artificial gastric juice resistance test: Take 0.1mL of bacterial suspension and add it into 0.9mL of artificial gastric juice (pH=3) and incubate at 37℃ for 3h; dilute the treated solution at 0h and 3h respectively and spread it on LMRS plate for live bacteria count. Calculate the survival rate of strains in artificial gastric juice and bile salt according to the following calculation formula:

[0081] Wherein, N1 represents the number of viable bacteria in the strain system after treatment, lg (CFU / mL); N0 represents the initial number of viable bacteria in the strain system, that is, the number of viable bacteria measured at 0h, lg (CFU / mL) Table 4. Tolerance results of Bifidobacterium breve in artificial gastric fluid (pH=3)

[0082] 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 had good tolerance and that Bifidobacterium breve BBr26 could survive in a low-acid gastric juice environment.

[0083] Experimental Example 6: Bifidobacterium breve BBr26 inhibits the growth of pathogenic bacteria Preparation of Escherichia coli ATCC25922 suspension: Escherichia coli ATCC25922 was inoculated into LB liquid culture medium at 2% (v / v), cultured at 37°C for 16-20h, and then the bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0084] Preparation of Salmonella Enteritidis ATCC14028 bacterial suspension: Salmonella Enteritidis ATCC14028 was inoculated into LB liquid culture medium at 2% (v / v), cultured at 37°C for 16-20h, and then the bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0085] Preparation of Staphylococcus aureus ATCC25923 bacterial suspension: Staphylococcus aureus ATCC25923 was inoculated into LB liquid culture 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.

[0086] Preparation of Streptococcus Parisi suspension: Bacteroides fragilis was inoculated into BHI blood plates containing 5% defibrinated sheep blood and cultured at 37°C for 16-20 h. The bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0087] 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 bacterial suspension concentration was adjusted to 10 8 CFU / mL.

[0088] 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.

[0089] Preparation of saprophytic Staphylococcus aureus suspension: Saprophytic Staphylococcus aureus was inoculated into LB liquid medium at 2% (v / v), cultured at 37°C for 16-20 h, and then the concentration of the bacterial suspension was adjusted to 10 8 CFU / mL.

[0090] 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.

[0091] Antibacterial experiment: MRS broth 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) was cooled to about 55°C and mixed with the indicator bacteria suspension in a certain proportion to make the number of viable indicator bacteria above 10. 6 CFU / mL, and then quickly poured into the plate with the Oxford cup placed in advance. After the culture medium cooled and solidified, take out the Oxford cup and inject 200μL of the strain fermentation liquid (the number of viable bacteria is 10 8 CFU level), and the diameter of the inhibition zone was measured after overnight culture at 37°C.

[0092] Table 5. Inhibition of pathogen growth by Bifidobacterium breve BBr26

[0093] 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, Propionibacterium acnes, etc. Among them, 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.

[0094] Experimental Example 7: Study on the proliferation of Bifidobacterium breve BBr26 on macrophage RAW264.7 Activation of Bifidobacterium breve BBr26: The strain BBr26 was taken out from a -80°C refrigerator, inoculated in a LMRS anaerobic tube liquid culture medium at a rate of 2% (v / v), and cultured at 37°C to obtain a first-generation Bifidobacterium breve BBr26 bacterial solution; then inoculated in a LMRS anaerobic tube liquid culture medium at a rate of 2% (v / v), and cultured at 37°C for 16 hours to obtain a second-generation Bifidobacterium breve BBr26 bacterial solution for use.

[0095] RAW264.7 cell culture: After taking out RAW264.7 cells from the ultra-low temperature refrigerator for recovery, use DMEM complete medium containing 1% non-essential amino acids, 1% penicillin-streptomycin and 10% fetal bovine serum, and place them in a constant temperature incubator with 5% CO2 at 37°C for culture. Change the medium every other day, and when the cell confluence reaches more than 80%, subculture. Wash twice with sterile PBS, add 2mL of culture medium, gently scrape the cells with a cell scraper, centrifuge and discard the supernatant, resuspend with DMEM complete culture medium, and subculture at a ratio of 1:3.

[0096] The concentration of RAW264.7 cells was adjusted to 1 × 10 4 cell / mL, inoculated in 96-well plates at 100μL / well, and cultured for 24h. Then the supernatant in the well plate was aspirated and washed twice with PBS. The experiment was divided into a blank group and a BBr26 group. The blank group: only 100μL DMEM culture medium was added to the plate wells, placed in a 5% CO2 incubator at 37℃ and cultured for 24h; the BBr26 group: 100μL bacterial suspension was added to the well plate (the bacterial suspension with good viable bacteria was washed twice with PBS, resuspended with DMEM, and diluted according to the ratio of bacterial volume: cell volume of 100:1), and cultured for 24h. After the incubation and culture, the supernatant in the well plate was aspirated, washed once with PBS, and 100μL of DMEM culture medium containing 10% CCK-8 was added. It was incubated at 37℃ in the dark for 1h, and the absorbance value (OD value) of each well at a wavelength of 450nm was measured with an enzyme marker.

[0097] The cell viability calculation method is CV=OD1 / OD2×100%.

[0098] 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.

[0099] Table 6. Results of the experiment on the proliferation of RAW264.7 cells by Bifidobacterium breve BBr26

[0100] Probiotics can enhance the activity of monocytes and macrophages, increase the content of reactive oxygen and reactive nitrogen, and phagocytic ability by inducing them. They can also process macrophage antigens and present them to T and B lymphocytes, causing humoral immunity and cellular immunity, thereby regulating the body's nonspecific and specific immune functions. The experiment on the proliferation of RAW264.7 macrophages by Bifidobacterium breve BBr26 found that the proliferation rate of RAW264.7 macrophages by Bifidobacterium breve BBr26 was as high as 178.32%, which was significantly higher than the common immunomodulatory strain Lactobacillus rhamnosus LGG, indicating that Bifidobacterium breve BBr26 has the function of immunomodulation.

[0101] Experimental Example 8: Verification of the effect of Bifidobacterium breve BBr26 in relieving atopic dermatitis in mice 1. Atopic dermatitis modeling in mice Forty healthy Balb / c mice aged 6 weeks were randomly divided into 4 groups, with 10 mice in each group. The 4 groups were blank group (Control), model group (DNFB) administered with 2,4-dinitrofluorobenzene (DNFB), positive drug group (KT) administered with ketotifen, and intervention group administered with Bifidobacterium breve BBr26. The experiment lasted for three weeks: the first week was the adaptation period for mice; gavage began in the second week until the end of the experiment.

[0102] (1) Bifidobacterium breve BBr26 intervention group: Oral administration of Bifidobacterium breve BBr26 bacterial solution, 0.2 mL of bacterial solution (1×10 10 CFU / mouse / time) by gavage; (2) Positive drug intervention group: 0.2 mL of ketotifen (KT) solution was administered intragastrically (the total amount of drug administered in a single intragastrical gavage was 1 mg / kg based on the mouse's body weight) per mouse per time. (3) The blank control group (NC) and the model group (MC) were not treated with B. breve BBr26 bacterial solution or ketotifen (KT) by oral gavage, and only an equal amount of normal saline was gavaged 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 back 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 back 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 every three days for a total of 4 times. The right ears of the blank group mice were only coated with an equal amount of acetone / olive oil matrix solution as a control; all groups were allowed to drink water and eat freely. The ear thickness of the six groups of mice was measured with a vernier caliper before the modeling began. After the modeling was completed, blood was collected and the mice were killed. The mouse serum was collected to measure the mouse-related inflammatory indicators; the mouse spleen and thymus were collected to measure the mouse-related organ index; the mouse ear thickness was measured, and the mouse back skin tissue was extracted for subsequent determination of related indicators.

[0103] Effect of Bifidobacterium breve BBr26 on ear thickness in mice with atopic dermatitis 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 2As shown in the figure, as the modeling time prolonged, the ear thickness of the blank control group remained basically unchanged, and the ear thickness of the model group (DNFB), BBr26 intervention group, and positive drug group (KT) mice was significantly (p<0.01) higher than that of the blank control group (Control), indicating that the atopic dermatitis mouse modeling was successful. The ear thickness of the right ear of the BBr2 pre-group mice was significantly (p<0.01) lower than that of the model group and the positive drug group coated with 0.5% DNFB solution, indicating that Bifidobacterium breve BBr26 can alleviate the allergic symptoms of atopic dermatitis mice.

[0104] Effects of Bifidobacterium breve BBr26 on inflammatory infiltration in the skin tissue of mice with atopic dermatitis After the experiment, the exposed skin tissues from the backs of mice in each group were extracted and stored in tissue freezing solution for H&E staining and section preparation. Figure 3 As shown, compared with the blank group (Control), the back skin thickness and inflammatory infiltration of mice in the model group (DNFB) were significantly increased; Bifidobacterium breve BBr26 significantly reduced the increase in mouse skin thickness and alleviated skin inflammatory infiltration; it can be seen that Bifidobacterium breve BBr26 can significantly improve skin inflammation in mice with atopic dermatitis.

[0105] 4. Mast Cell Staining in Atopic Dermatitis Mice The 5µm thick slices were baked, dehydrated with alcohol, stained with mast cell staining solution, differentiated, and sealed to make pathological sections. Three different fields of view were selected under an optical microscope to count the number of mast cells. After the modeling was completed, blood was collected and the mice were killed. The skin of the depilated area on the back of the mice was taken for histopathological analysis. The histopathological sections of the back skin of the mice were stained with toluidine blue dye. It can be seen from 4 that the infiltration of mast cells in the skin tissue was reduced. At the same time, the number of mast cells produced was reduced, and the skin Bifidobacterium breve BBr26 was inhibited, which significantly inhibited tissue inflammation.

[0106] 5. Determination of thymus index and spleen index of atopic dermatitis mice 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 shown in Figure 5 , Figure 6 Compared 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.

[0107] 6. Determination of inflammatory factors in serum of mice with atopic dermatitis After the modeling, blood was collected and the mice were killed. Blood was collected from the eyeballs of the mice and centrifuged at 3000 rpm / min×15min. The upper serum was collected and the levels of IL-4, IL-13, IL-10, and INF-γ were measured using an enzyme-linked immunosorbent assay kit. The core mechanism of atopic dermatitis is an immune abnormality dominated by Th2-type reactions. After invading the skin barrier, allergens are taken up by antigen-presenting cells DC and LC, and then DC presents the allergens to Th0 cells. Under the action of cytokine IL-4, Th0 cells differentiate and proliferate into Th2. In this process, Th2 cell-specific nuclear transcription factor GATA3 plays a decisive role. A large number of Th2 cells secrete cytokines IL-4, IL-5, and IL-13, forming an excessive Th2-type immune response. The excessive secretion and expression of these Th2-type cytokines leads to a decrease in the expression of loricrin and involucrin, which maintain the integrity of the skin barrier, thereby exacerbating the destruction of the skin barrier.

[0108] 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.

[0109] IL-10 helps to inhibit Th2 immune response and restore Th1 / Th2 immune balance. The results of the measurement of IL-10 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.

[0110] 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 the expression of IL-4 receptor.

[0111] VII. Determination of IgE Content in Atopic Dermatitis Mice The core immune mechanism of atopic dermatitis is the 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 measurement 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 having the ability to inhibit hyaluronidase activity and relieve allergies ( Bifidobacterium breve )BBr26 ， It is characterized in that its deposit number is CGMCC NO.32424.

2. Bifidobacterium breve with a deposit number of CGMCC NO.32424 ( Bifidobacterium breve ) Application of BBr26 in the preparation of products for alleviating atopic dermatitis, and / or foods for enhancing immune function, and / or antibacterial agents.

3. The Bifidobacterium breve (CGMCC NO.32424) according to claim 2. Bifidobacterium breve ) Application of BBr26 in the preparation of a product for relieving atopic dermatitis, and / or a food for enhancing immune function, and / or an antibacterial agent, characterized in that: 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; And / or, the immunity 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; And / or, the regulating cytokine levels includes: reducing the levels of IL-4 and IL-13, and increasing the levels of IL-10 and INF-γ; And / or, the antibacterial spectrum of the antibacterial agent includes: Escherichia coli, Salmonella enteritidis, Staphylococcus aureus, Streptococcus anginosus, Streptococcus Parisi, Streptococcus pasteurianus, Staphylococcus saprophyticus, Propionibacterium acnes.

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

5. A product for relieving atopic dermatitis according to claim 4, characterized in that: Also includes: Auxiliary materials.

6. A food for enhancing immune function, comprising: Active ingredient; characterized in that the active ingredient includes: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

7. The food for enhancing immune function according to claim 6, characterized in that: Also includes: Auxiliary materials.

8. An antibacterial agent comprising: Active ingredient; characterized in that the active ingredient includes: Bifidobacterium breve ( Bifidobacterium breve )BBr26.

9. An antibacterial agent according to claim 8, characterized in that: Also includes: Auxiliary materials.

10. An in vitro antibacterial method, characterized in that: The Bifidobacterium breve (CGMCC NO.32424) was used. Bifidobacterium breve )BBr26 is antibacterial.

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