Bifidobacterium bifidum and application thereof in relieving ovalbumin-induced food allergy

Abstract

The invention discloses bifidobacterium bifidum and application of the bifidobacterium bifidum in relieving ovalbumin-induced food allergy, and belongs to the technical field of microorganisms. According to the invention, a strain of bifidobacterium bifidum CCFM1063 is screened out, and the bifidobacterium bifidum CCFM1063 has the effects of relieving weight loss and allergic symptoms of mice caused by OVA; according to the present invention, with the application of the loratadine, the high mMCP-1 level caused by OVA can be significantly reduced, the IgE level can be reduced, the mRNA level of the jejunum allergy related cytokines such as TSLP, IL-4 and IL-33 can be significantly reduced, and the partial effect is superior to the loratadine; when the bifidobacterium bifidum CCFM1063 is combined with breast milk oligosaccharide 3 '-SL, the bifidobacterium bifidum CCFM1063 has a synergistic effect on improving food allergy symptoms, and the effect of the bifidobacterium bifidum CCFM1063 is obviously superior to that of a single strain.

Description

Technical Field

[0001] The invention relates to Bifidobacterium bifidum and application thereof in alleviating food allergy induced by ovalbumin, and belongs to the technical field of microorganisms and the technical field of medicine. Background Art

[0002] Allergic diseases are increasing worldwide, with airway and food allergies being the most common, especially in infants and young children. Food allergies usually occur when the body is hypersensitive to certain proteins when ingesting food. For infants and young children whose gastrointestinal function is not yet fully developed, food allergies may cause chronic non-infectious diarrhea, which has a long course and is prone to recurrence, seriously affecting growth, development and health.

[0003] Currently, the common methods of avoiding or treating food allergies are avoiding the consumption of foods containing corresponding allergens and their products, removing the corresponding allergens from foods, and using anti-allergic drugs for clinical treatment. However, these methods cannot fundamentally alleviate or control food allergies. Clinical treatment drugs are mainly specific immunotherapy to induce the body's tolerance to allergens. When the host is exposed to the allergens again, it can induce allergic reactions of varying degrees.

[0004] The pathogenesis of food allergies involves complex immune mechanisms triggered by exposure to certain food proteins. When these proteins or allergens that are mistakenly considered harmful by the body are absorbed by antigen-presenting cells, they stimulate B cells to produce allergen-specific immunoglobulin E (IgE) antibodies. After subsequent exposure to the same allergen, IgE antibodies that have bound to high-affinity IgE receptors on mast cells and basophils recognize allergens, triggering mast cells to release various mediators including mMCP-1, leading to allergic symptoms such as anaphylactic shock. Therefore, IgE and mMCP-1 are important parameters for judging the severity of food allergies. Although the pathogenesis of food allergies is still unclear, intestinal microecological imbalance and immune dysfunction are at the core. Some beneficial microorganisms in the intestine can improve intestinal peristalsis, prevent pathogenic bacteria from colonizing in the intestine, and maintain the balance of intestinal flora. Probiotics can maintain the regulatory effect of the intestinal mucosal immune system through various mechanisms, regulate type 2 immunity, and promote intestinal barrier functions. Therefore, the discovery of probiotics or probiotics combined with prebiotics that can improve food allergy symptoms may solve the problems of recurrence in food allergy treatment. Currently, few bifidobacteria have been discovered that can regulate IgE and mMCP-1 levels in food allergic individuals and enhance the alleviating effect of human milk oligosaccharides on food allergies. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide a strain that can be used to prevent and / or treat food allergies; and to provide a strain that can be compounded with oligosaccharides for the treatment of food allergies to achieve a synergistic promoting effect.

[0006] In order to achieve the above objectives, the present invention provides the use of Bifidobacterium bifidum CCFM1063 in the preparation of medicines for alleviating food allergies.

[0007] In one embodiment, the relief of food allergy comprises at least one of the following aspects:

[0008] (1) Improve weight loss caused by food allergies;

[0009] (2) reduce the expression levels of IgE and mMCP-1 in individuals with food allergies;

[0010] (3) regulating the expression levels of individual inflammatory factors;

[0011] (4) Improve jejunal tissue lesions.

[0012] In one embodiment, the drug further comprises human milk oligosaccharides; optionally, the human milk oligosaccharides include 3'-sialyllactose.

[0013] In one embodiment, the content of human milk oligosaccharides in the drug is not less than 4×10 -4 g / g.

[0014] In one embodiment, the viable count of Bifidobacterium bifidum CCFM1063 in the drug is not less than 1×10 9 CFU / g.

[0015] In one embodiment, the Bifidobacterium bifidum CCFM1063 is in the form of bacterial liquid, lyophilized powder or fermentation composition.

[0016] The present invention also provides a medicine for alleviating food allergies, wherein the medicine contains Bifidobacterium bifidum CCFM1063; optionally, the medicine also contains human milk oligosaccharides.

[0017] The present invention also provides a health product for enhancing immunity, wherein the medicine contains Bifidobacterium bifidum CCFM1063; optionally, the medicine also contains human milk oligosaccharides.

[0018] The present invention also provides the use of Bifidobacterium bifidum CCFM1063 in enhancing the efficacy of food allergy drugs, wherein the drug contains human milk oligosaccharides as a main active ingredient.

[0019] In one embodiment, the medicine contains a synbiotic prepared by mixing Bifidobacterium bifidum CCFM1063 and human milk oligosaccharides; the Bifidobacterium bifidum CCFM1063 includes a bacterial liquid, a lyophilized powder or a fermentation composition of Bifidobacterium bifidum CCFM1063.

[0020] Beneficial effects:

[0021] 1. The present invention provides Bifidobacterium bifidum CCFM1063 that can alleviate the pathological characteristics of food allergic mice and thus alleviate food allergies. After the bifidobacterium provided by the present invention acts on food allergic mice, the allergic reactions of the mice can be significantly alleviated. Specifically, compared with the modeling group, the Bifidobacterium bifidum CCFM1063 provided by the present invention can: (1) improve the weight loss of food allergic mice; (2) significantly improve the serum OVA-specific IgE level of food allergic mice; (3) reduce the serum mMCP-1 level of food allergic mice; (4) significantly reduce the serum IL-4 level of food allergic mice; (5) improve the degree of jejunal tissue lesions in food allergic mice; and (6) significantly reduce the mRNA levels of Th2 cell-related cytokines IL-4 and epithelial cell factors IL-33 and TLSP in the jejunal tissue of food allergic mice.

[0022] 2. Bifidobacterium bifidum CCFM1063 and human milk oligosaccharides play a synergistic role in alleviating food allergies. Combining Bifidobacterium bifidum CCFM1063 with human milk oligosaccharides to prepare synbiotics can effectively improve the alleviating effect on food allergies. Compared with the model group, the allergy symptom score was reduced by 60%, the mMCP-1 level was reduced by 51.7%, the OVA-sIgE level was reduced by 38.8%, and the IL-4 level was reduced by 43.4%.

[0023] Therefore, the Bifidobacterium bifidum CCFM1063 of the present invention has great application prospects in the preparation of products (such as food or medicine, etc.) for preventing and / or treating food allergies.

[0024] Bifidobacterium bifidum has been included in the "List of Bacteria that Can Be Used in Food" issued by the Ministry of Health. In addition, as a probiotic, Bifidobacterium bifidum has been proven to have the following functions: (1) Maintaining the balance of normal intestinal bacterial flora, inhibiting the growth of pathogens, and preventing diseases such as constipation, diarrhea and gastrointestinal disorders; (2) Anti-tumor; (3) Synthesizing nutrients such as vitamins and amino acids in the intestine; (4) Improving the body's absorption of calcium ions; (5) Lowering blood cholesterol levels and preventing and treating hypertension; (6) Improving the lactose tolerance of dairy products and increasing digestibility; (7) Enhancing human immune function; (8) Anti-aging; (9) Enhancing the body's nonspecific and specific immune responses; (10) Controlling the effect of toxinemia in the body; (11) Improving the host's tolerance to radiation. Therefore, the Bifidobacterium bifidum CCFM1063 screened and obtained by the present invention is healthy, has no toxic side effects and has certain benefits to the human body. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 : Changes in body weight percentage of experimental mice in different groups.

[0026] Figure 2 : Allergy symptom scores of experimental mice in different groups.

[0027] Figure 3 : The levels of serum OVA-specific IgE, mast cell protease 1 (mMCP-1), and IL-4 in different groups of experimental mice. In the figure: *: P<0.05, **: P<0.01, ***: P<0.001.

[0028] Figure 4 :Pathological sections of jejunum tissues of experimental mice in different groups.

[0029] Figure 5 :Cytokine mRNA levels in jejunal tissues of experimental mice in different groups. In the figure: *: P<0.05, **: P<0.01, ***: P<0.001.

[0030] Figure 6 :Bifidobacterium bifidum CCFM1063 and 3'-SL synergistically alleviate food allergy in mice. A. Allergy score; B. Mast cell protease 1 (mMCP-1); C. Serum OVA-specific IgE; D. Serum IL-4 level. In the figure: *: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.0001. DETAILED DESCRIPTION

[0031] The present invention will be better understood through the following examples.

[0032] In the present invention, unless otherwise specified, "%" or percentage used to describe concentration or ratio are all weight percentages.

[0033] The present invention relates to the following culture medium:

[0034] MRS liquid culture medium: 10 g tryptone, 10 g beef extract, 5 g yeast powder, 20 g glucose, 2 g diammonium hydrogen citrate, 5 g sodium acetate, 2 g dipotassium hydrogen phosphate, 0.5 g magnesium sulfate heptahydrate, 0.25 g manganese sulfate monohydrate, 1 mL Tween 80 and 0.5 g cysteine, add water to 1000 mL.

[0035] MRS solid medium is obtained by adding 1.5% agar based on the total weight of liquid medium.

[0036] The BALB / c female mice involved in the following examples were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., and the OVA involved in the following examples was purchased from Sigma-Aldrich.

[0037] The preparation method of the OVA solution involved in the following embodiments is as follows:

[0038] OVA sensitization solution: Each mouse was intraperitoneally injected with 0.2 mL of sensitization solution, which was a mixture of 0.1 mL of normal saline containing 50 μg of OVA and 0.1 mL of alum adjuvant containing 2 mg of aluminum hydroxide.

[0039] OVA provocation solution: Each mouse was intragastrically administered with 0.2 mL of provocation solution, which was physiological saline containing 50 mg OVA.

[0040] The Bifidobacterium bifidum CCFM1063 involved in the following examples has been disclosed in patent CN110331119B.

[0041] Example 1: Cultivation of Bifidobacterium bifidum CCFM1063

[0042] The specific steps are as follows:

[0043] Bifidobacterium bifidum CCFM1063 was inoculated into MRS liquid culture medium and cultured at 37°C for 24 hours. The bacterial liquid was then transferred into fresh MRS liquid culture medium at a 2% inoculation rate and cultured under the same conditions for 24 hours. The bacteria were centrifuged at 8000 r / min for 15 minutes, washed with 0.9% saline and centrifuged again at 8000 r / min for 10 minutes. The bacteria were collected and resuspended with 30% (m / v) sucrose solution to prepare resuspensions, which were frozen at -80°C for use.

[0044] Preparation of bacterial suspension for oral administration: When Bifidobacterium bifidum CCFM1063 is used for oral administration to mice, the prepared resuspension is taken out from -80°C, centrifuged at 4°C, 8000r / min for 10min, the supernatant is discarded, and the suspension is resuspended with 0.9% saline to obtain a bacterial suspension for oral administration.

[0045] Example 2: Effect of Bifidobacterium bifidum CCFM1063 on the alleviation of weight loss in mice with food allergy

[0046] The specific steps are as follows:

[0047] Four-week-old SPF BALB / c female mice were divided into four groups, namely, normal group (Control), model group (OVA), positive control group (OVA+Loratadine), and Bifidobacterium bifidum group (OVA+CCFM1063);

[0048] Each group consisted of 8 animals, housed in the Experimental Animal Center of Jiangnan University, fed with ordinary feed, with a constant temperature of 21-26°C, humidity of 40-70%, noise less than or equal to 60dB, and animal illumination of 15-20LX (all animal experimental procedures were reviewed and approved by the Animal Welfare and Ethics Management Committee of Jiangnan University).

[0049] The experimental period was 7 weeks in total. The first week was the adaptation period. The mice were weighed after the adaptation period. During the adaptation period, they were fed with Co60 irradiated growth and breeding feed for experimental mice (Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd.).

[0050] Modeling process: Modeling began in the second week (counted as day 0), mice were weighed at the same time every week, and OVA sensitization solution was injected intraperitoneally on days 0 and 14 for sensitization; OVA challenge solution was gavaged from day 28, 0.2 mL per mouse, once every 2 days, for a total of 6 times, until the last OVA challenge solution gavage on day 43, and mice were weighed the day after each gavage. The mice were killed after the last weighing of the mice 1 hour after the last challenge.

[0051] Normal group (Control): 0.2 mL of 0.9% saline was administered intragastrically every day.

[0052] Model group (OVA): During the modeling process, mice were intragastrically administered with 0.2 mL of 0.9% saline every day.

[0053] Positive control group (OVA+Loratadine): During the modeling process, mice in the experimental group were intragastrically administered with 20 mg / kg WB of Loratadine every day.

[0054] Bifidobacterium bifidum CCFM1063 (OVA+CCFM1063) group: During the modeling process, mice in the experimental group were gavaged with 0.2 mL of bacterial solution every day (the number of viable bacteria per gavage was 1×10 9 The preparation method of bacterial suspension was the same as that in Example 1.

[0055] The results are as follows Figure 1 As shown: During the experiment, the percentage changes in the body weight of mice in different groups (with day 0 as 100%). Among them, after the start of the OVA oral challenge (28 days from the start of the experiment), the weight change of the model group showed a significant downward trend, and Bifidobacterium bifidum CCFM1063 effectively alleviated the weight loss of mice caused by food allergies, and had a similar alleviating effect as the existing loratadine. At the end of the experiment, the average weight of mice in the Bifidobacterium bifidum CCFM1063 group was 17.2g, which was 1.1 times higher than that of the loratadine group, indicating that Bifidobacterium bifidum CCFM1063 is better than drugs in alleviating food allergies caused by OVA.

[0056] Example 3: Effect of Bifidobacterium bifidum CCFM1063 on allergic symptoms in food allergic mice

[0057] The experimental methods and bacterial activation are as described above.

[0058] After the last oral administration of OVA solution, the allergic symptoms of mice were scored according to the allergic symptom scoring table. Among them, "no allergic symptoms" was recorded as 0 points, "repeated scratching of ears and mouth, scratching of ear canals with hind legs, scratching marks on tail, and rough hair" was recorded as 1 point, "reduced activity, reduced food intake, and messy hair" was recorded as 2 points, "long-term immobility, accelerated breathing rate, messy and dull hair, and erect hair" was recorded as 3 points, "protruding eyeballs, conjunctival congestion, no response to stimulation, tremors, convulsions, and cyanosis of lips" was recorded as 4 points, and "death" was recorded as 5 points. The scoring results are shown in the table. Figure 2 .

[0059] Depend on Figure 2 It can be seen that the mice in the normal group had smooth and shiny hair and no allergic symptoms; the mice in the model group had messy and dull hair and had piloerection; Bifidobacterium bifidum CCFM1063 can improve the allergic symptoms of mice caused by food allergies, and has a similar effect to loratadine.

[0060] Example 4: Alleviating effect of Bifidobacterium bifidum CCFM1063 on serum OVA-specific IgE, mMCP-1 and IL-4 levels in food allergic mice

[0061] The experimental methods and bacterial activation are as described above.

[0062] After removing the eyeballs to collect blood and killing the mice, the blood was allowed to stand for 2 hours and then centrifuged at 3000r / min to collect the mouse serum. The levels of OVA-specific IgE, mast cell protease (mMCP-1) and allergy-related cytokine IL-4 in the mouse serum were determined by ELISA kits. The results are shown in Figure 3 shown.

[0063] Depend on Figure 3 It can be seen that compared with the model group, after mice were gavaged with the bacterial solution of Bifidobacterium bifidum CCFM1063, the content of OVA-specific IgE in the serum decreased, which was similar to the positive control group. The level of mast cell protease-1 (mMCP-1) in the serum of the Bifidobacterium bifidum CCFM1063 group was also significantly lower than that of the model group (p<0.001), a decrease of 33.9%, similar to loratadine (a decrease of 30.6%); in addition, the level of IL-4, a cytokine associated with Th2 cells in the serum, was also slightly reduced in the Bifidobacterium bifidum CCFM1063 group. The above data show that the strain Bifidobacterium bifidum CCFM1063 in the present invention can reduce the high mMCP-1 level caused by OVA.

[0064] Example 5: The alleviating effect of Bifidobacterium bifidum CCFM1063 on the lesion degree of jejunum tissue in mice with food allergy. The experimental method and strain activation are as mentioned above.

[0065] The mice were killed 1 hour after the last oral administration of OVA solution. The mouse jejunal tissue was fixed with 4% paraformaldehyde, and then washed, dehydrated, transparent, waxed, embedded, sliced, spread, glued, baked, stained with hematoxylin-eosin (HE), differentiated, rinsed, counterstained, dehydrated, transparent and sealed to make jejunal HE stained sections. Pathological section scanner was used to record micrographs. The results of jejunal HE pathological sections are shown in Figure 4 .

[0066] Depend on Figure 4 It can be seen that the villi of the small intestine of the jejunum of the normal group of mice were complete and neatly arranged, and the lamina propria was tight and not loose;

[0067] The intestinal villi of the jejunum of the model group mice were broken and arranged in disorder, with inflammatory cell infiltration and loose lamina propria;

[0068] Compared with the model group, the integrity, arrangement density, inflammatory cell infiltration and looseness of the lamina propria of the jejunum of mice in the Bifidobacterium bifidum CCFM1063 group were alleviated, which had a similar improvement effect as loratadine.

[0069] Example 6: Effects of Bifidobacterium bifidum CCFM1063 on the levels of inflammatory factors in the jejunum tissue of food allergic mice The experimental method and strain activation are as described above.

[0070] Mice were killed 1 hour after the last oral administration of OVA solution. The jejunal tissue of mice was taken and quickly frozen with liquid nitrogen. 50 mg of jejunal tissue was weighed, and the total RNA in the jejunal tissue was extracted by TriZol method. The RNA was reverse transcribed into cDNA using HiScriptⅢAll-in-one RTSuperMix, and the target gene was amplified by real-time quantitative polymerase chain reaction (qRT-PCR). The qRT-PCR system was 2ⅹSybGreen 5μL, 0.2μL of forward and reverse primers, 1μg of template cDNA, and it was made up to 20μL with water. The qRT-PCR amplification conditions were 95℃5min, 95℃30s, 60℃30s, 72℃2min, 35 cycles, and 72℃10min. The sequences of primers for related genes are shown in Table 1. The expression changes of various cytokine genes relative to the housekeeping gene (β-actin) were calculated according to the formula: gene relative expression level = 2-ΔΔCt. The results are shown in Figure 5 .

[0071] Table 1 Primer sequences

[0072]

[0073]

[0074] Depend on Figure 5 It can be seen that after mice were gavaged with Bifidobacterium bifidum CCFM1063, the levels of cytokines IL-4, IL-33 and TSLP mRNA in jejunal tissue were significantly lower than those in the model group (p<0.05), indicating that the strain Bifidobacterium bifidum CCFM1063 in the present invention can inhibit intestinal Th2 response.

[0075] Overall, it was shown that Bifidobacterium bifidum CCFM1063 could improve OVA-induced allergic reactions by significantly downregulating the level of mMCP-1, and had a good regulatory effect on intestinal tissue pathological damage and allergy-related cytokines, ultimately manifesting as a significant alleviating effect on OVA sensitization.

[0076] The above experimental results show that Bifidobacterium bifidum CCFM1063 can effectively improve the symptoms of weight loss and allergies in mice. ELISA results showed that Bifidobacterium bifidum CCFM1063 improved the key indicators of OVA-induced sensitization, including significantly downregulating the level of mMCP-1. The results of intestinal pathological sections showed that Bifidobacterium bifidum CCFM1063 could significantly improve the pathological damage of the jejunum of mice caused by OVA, effectively increasing the length of villi and reducing the depth of crypts. qPCR results showed that Bifidobacterium bifidum CCFM1063 could also regulate the homeostasis of the jejunal immune system, including significantly downregulating the levels of proinflammatory cytokines related to intestinal allergies.

[0077] In summary, Bifidobacterium bifidum CCFM1063 can significantly improve OVA-induced food allergy in mice. It can be used to prepare medicines or health products for the prevention and treatment of food allergies, or to produce foods that are beneficial to intestinal health, such as probiotic beverages, sour soy milk, fermented jelly, fermented tea beverages or dairy products (such as yogurt, cheese products, lactic acid bacteria, milk powder), etc.

[0078] Example 7: The alleviating effect of Bifidobacterium bifidum CCFM1063 combined with 3'-SL on food allergy in mice

[0079] The experimental method and strain activation are shown in Example 1-2, except that two new groups were added: the human milk oligosaccharide 3'-sialyllactose 3'-Sialyllactose (3'-SL group) (400 mg / kg / d) group, and the synbiotic combination of Bifidobacterium bifidum CCFM1063 and 3'-SL (400 mg / kg / d) (3'-SL+CCFM1063 group). During the modeling process, the 3'-SL group was gavaged with 400 mg / kg 3'-SL once a day; the 3'-SL+CCFM1063 group was gavaged with 400 mg / kg 3'-SL and 0.2 mL of Bifidobacterium bifidum CCFM1063 bacterial solution once a day (the number of viable bacteria in a single gavage was 1×10 9 CFU). After the last oral administration of OVA solution, the allergic symptoms of mice were scored according to the allergic symptom scoring table, and the serum OVA-specific IgE and mMCP-1 levels were detected.

[0080] Figure 6 The results showed that both CCFM1063 and synbiotic groups could regulate food allergies, but the synbiotic group had a better effect than CCFM1063 and 3'-SL groups, and there was a significant difference, indicating that CCFM1063 and 3'-SL had a synergistic effect, which was better than the single CCFM1063 and breast milk oligosaccharide groups. Compared with the model group, the allergy symptom score of the synbiotic group decreased by 60%, the mMCP-1 level decreased by 51.7%, the OVA-sIgE level decreased by 38.8%, and the IL-4 level decreased by 43.4%.

[0081] In summary, the combination of Bifidobacterium bifidum CCFM1063 and 3'-SL in the embodiment of the present invention can significantly reduce the levels of specific antibody IgE and serum mMCP-1 and IL-4 in food allergic mice, thereby alleviating allergic reactions. Therefore, the synbiotic combination of Bifidobacterium bifidum CCFM1063 and 3'-SL can be used to alleviate food allergic reactions.

[0082] Patent CN114949006 A discloses Bifidobacterium breve M_16V that can alleviate food allergies. However, the above patent does not include OVA-sIgE and mMCP-1, two key indicators for evaluating food allergies in OVA-induced food allergies. In addition, the IL-4 level in the serum detected by the patent did not change significantly. In another study, it was found that Bifidobacterium breve M_16V alone reduced the allergic symptom score and mMCP-1 level in mice, but the difference was not significant compared with the model group. Only after the bacteria were combined with oligosaccharides (scFOSlcFOS) did the allergy score and mMCP-1 level significantly reduce (P < 0.05) (van Esch BC, Abbring S, Diks MA, Dingjan GM, Harthoorn LF, Vos AP, Garssen J. Post-sensitization administration of non-digestible oligosaccharides and Bifidobacterium breve M-16V reduces allergic symptoms in mice. Immun Inflamm Dis. 2016, 4(2): 155-165.). The Bifidobacterium bifidum CCFM1063 in this patent significantly reduced the mouse allergy score (P < 0.05), the level of mast cell protease mMCP-1 (P < 0.05), and after subsequent compounding with breast milk oligosaccharides, the mouse allergy score dropped more significantly (P < 0.001), and at the same time, the OVA-specific IgE, mMCP-1 (P < 0.001), and IL-4 levels were significantly reduced. Therefore, the Bifidobacterium bifidum CCFM1063 described in the present invention, especially after compounding with breast milk oligosaccharides 3'-SL, can simultaneously regulate the abnormal levels of specific IgE, mMCP-1, and IL-4 caused by food allergies, and the effect of improving food allergies is significantly better than the compounding of Bifidobacterium breve M_16V and oligosaccharides.

[0083] Example 8: Preparation of freeze-dried powder of Bifidobacterium bifidum CCFM1063

[0084] Bifidobacterium bifidum CCFM1063 is inoculated into the culture medium at an inoculum amount of 3% of the total mass of the culture medium, and cultured at 37°C for 18 hours to obtain a culture solution; the culture solution is centrifuged to obtain bacterial cells; the culture solution is centrifuged to obtain bacterial cells; the bacterial cells are washed three times with a phosphate buffer solution with a pH of 7.2 to 7.4, and then resuspended with a trehalose lyophilization protectant with a trehalose concentration of 100 g / L (the mass ratio of the lyophilization protectant to the bacterial cells is 2:1) to obtain a resuspension; the resuspension is freeze-dried by a vacuum freezing method to obtain freeze-dried powders of Bifidobacterium breve CFM1400; wherein the culture medium contains water accounting for 87.7% of the total mass of the culture medium, 10% of enzyme-hydrolyzed skim milk, 0.5% of glucose, 1.5% of trypsin and 0.3% of yeast extract solution; the pH of the culture medium is 6.8.

[0085] Take 1×10 9 CFU of Bifidobacterium bifidum CCFM1063 freeze-dried powder, reconstituted with water and then gavaged into OVA-induced food allergic mice, can effectively alleviate the symptoms of sensitized mice.

[0086] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. Application of Bifidobacterium bifidum CCFM1063 in the preparation of medicines for alleviating food allergies.

2. The use according to claim 1, characterized in that The relief of food allergy includes at least one of the following aspects: (1) Improve weight loss caused by food allergies; (2) reduce the expression levels of IgE and mMCP-1 in individuals with food allergies; (3) regulating the expression level of inflammatory factors; (4) Improve jejunal tissue lesions.

3. The use according to claim 1 or 2, characterized in that: The medicine further comprises human milk oligosaccharides; optionally, the human milk oligosaccharides include 3'-sialyllactose.

4. The use according to claim 3, characterized in that The content of human milk oligosaccharides in the drug is not less than 4×10 -4 g / g.

5. The use according to claim 4, characterized in that The number of Bifidobacterium bifidum CCFM1063 in the drug is not less than 1×10 9 CFU / g.

6. The use according to claim 5, characterized in that The Bifidobacterium bifidum CCFM1063 is in the form of bacterial liquid, freeze-dried powder or fermentation composition.

7. A medicine for alleviating food allergies, characterized in that: The medicine contains Bifidobacterium bifidum CCFM1063; optionally, the medicine also contains human milk oligosaccharides.

8. A health product for enhancing immunity, characterized in that: The medicine contains Bifidobacterium bifidum CCFM1063; optionally, the medicine also contains human milk oligosaccharides.

9. Use of Bifidobacterium bifidum CCFM1063 in enhancing the efficacy of food allergy drugs, characterized in that: The medicine uses human milk oligosaccharides as a main active ingredient.

10. The use according to claim 9, characterized in that The medicine contains a synbiotic prepared by mixing Bifidobacterium bifidum CCFM1063 and human milk oligosaccharides; the Bifidobacterium bifidum CCFM1063 includes a bacterial liquid, a lyophilized powder or a fermentation composition of Bifidobacterium bifidum CCFM1063.

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