A probiotic composition for improving allergy

Abstract

The application provides an allergy-improving probiotic composition, and belongs to the field of microbial preparations.The probiotic composition comprises Bacillus coagulans powder, Bacillus coagulans postbiotic, fructooligosaccharide, lactitol, pin cherry powder and stachyose.The application optimizes the application form of Bacillus coagulans, and further compounding other raw materials to prepare a composition which can effectively relieve allergy, and as a whole, has a significant effect on an animal model.

Description

Technical Field

[0001] This invention belongs to the field of microbial preparations, and specifically relates to a probiotic composition for improving allergies. Background Technology

[0002] Allergy is an abnormal immune response that primarily results in tissue and cell damage or physiological dysfunction. When an allergen enters the body, it sensitizes the body and triggers the production of antibodies. When the allergen re-enters the body, it binds to the existing antibodies, causing a series of allergic reactions. Allergy is a major representative of type I hypersensitivity reactions and is clinically broadly divided into allergic reactions and allergic diseases. The former is a systemic syndrome caused by an abnormal reaction to an allergen. The latter involves a specific organ or tissue, leading to a disease. Common allergic diseases include allergic asthma, allergic rhinitis, hay fever, and certain types of dermatitis.

[0003] Anti-allergy probiotics are beneficial to the body. These microorganisms can effectively improve allergic constitution, promote nutrient absorption, and enhance the body's immunity. For example, compound Lactobacillus acidophilus helps improve the imbalance of human immune function, regulate the body's immunity, enhance the body's resistance, and help prevent and improve allergy symptoms. The most common anti-allergy probiotics also include Lactobacillus acidophilus and Lactobacillus reuteri. Anti-allergy probiotics can effectively relieve allergy symptoms and have very few side effects on the body, which is one aspect that makes them superior to other anti-allergy drugs.

[0004] Epigenetics are a collective term for the metabolites of probiotics after processing, including the bacterial cells and metabolic products. Selected epigenetics retain high physiological activity even after high-temperature treatment or processing with gastrointestinal digestive fluids, and they offer various health benefits to the human body. Existing technologies CN117085046A and CN116769682A both disclose the allergic-relieving effects of probiotic epigenetics.

[0005] *Bacillus coagulans* belongs to the genus *Bacillus* in taxonomy. Its cells are rod-shaped, Gram-positive, terminally spore-forming, and non-flagellated. It decomposes sugars to produce L-lactic acid, making it a homolactic acid fermenting bacterium. Existing technology CN112755054A discloses the application of *Bacillus coagulans* in the prevention and / or treatment of allergic diseases, clearly demonstrating its allergic-relieving and therapeutic effects; CN114480218A discloses *Bacillus coagulans* with the ability to reduce inflammatory factors. However, existing technologies lack research on the postbiotics of these strains. Furthermore, the application of combinations of probiotics and related postbiotics in anti-allergy treatments is limited in existing technologies, indicating significant room for development. Summary of the Invention

[0006] To address the aforementioned problems, this invention provides a compound composition that optimizes the use of strains and auxiliary materials in the prior art, resulting in a product that can effectively control allergies.

[0007] On the one hand, the present invention provides a probiotic composition for improving allergies.

[0008] The probiotic composition includes: Bacillus coagulans powder, Bacillus coagulans postbiotic, fructooligosaccharides, lactitol, acerola cherry powder, and stachyose.

[0009] Preferably, the ratio of Bacillus coagulans powder to Bacillus coagulans postbiotic is 1-5:1.

[0010] Preferably, the probiotic composition contains 10 probiotics. 9 -10 10 CFU / g.

[0011] More preferably, the probiotic content in the Bacillus coagulans powder is 5 × 10⁻⁶. 9 CFU / g.

[0012] Specifically, by weight, it includes: 1.5-2.5 parts of Bacillus coagulans powder, 0.5-1.5 parts of Bacillus coagulans post-generic, 1-2 parts of fructooligosaccharides, 0.5-1 part of lactitol, 0.5-1.5 parts of acerola cherry powder, and 1-2 parts of stachyose.

[0013] More specifically, by weight, it includes: 2 parts of Bacillus coagulans powder, 1 part of Bacillus coagulans post-generic, 1 part of fructooligosaccharides, 0.5 parts of lactitol, 1 part of acerola cherry powder, and 2 parts of stachyose.

[0014] Preferably, the preparation method of the Bacillus coagulans metagener is ultrasonic disruption. In fact, those skilled in the art can also obtain the metagener using lysozyme, as disclosed in the metagener preparation method in patent application number CN202311365361.1.

[0015] More preferably, the method for preparing the Bacillus coagulans postbiotic is as follows:

[0016] Fermentation broth was obtained by inoculating 1%-5% of Bacillus coagulans into a culture medium, collecting the bacterial cells, and resuspending them in PBS at a concentration of 1×10⁻⁶. 9 -10×10 9 CFU / mL, inactivate; treat with 500-600W power for 10-20 min at a frequency of 20-30kHz, with ultrasonic treatment for 5 s and a 5 s interval.

[0017] On the other hand, the present invention provides a method for preparing a probiotic composition for improving allergies.

[0018] The preparation method includes:

[0019] (1) Preparation of Bacillus coagulans powder and Bacillus coagulans post-genetic agent;

[0020] (2) The probiotic composition is compounded according to the above-mentioned formula.

[0021] The preparation method of the Bacillus coagulans powder can be as follows: 1%-5% of Bacillus coagulans is inoculated into a fermentation medium and cultured for 24 hours to obtain a fermentation broth. The broth is then centrifuged at 10,000 rpm for 20 minutes to collect the bacterial cells, freeze-dried, and the powder is obtained.

[0022] Those skilled in the art can selectively adjust the above scheme to obtain a bacterial powder with the desired bacterial content.

[0023] The method for preparing the Bacillus coagulans postbiotic is as follows:

[0024] Fermentation broth was obtained by inoculating 1%-5% of Bacillus coagulans into a culture medium, collecting the bacterial cells, and resuspending them in PBS at a concentration of 1×10⁻⁶. 9 -10×10 9 CFU / mL, inactivate; treat with 500-600W power for 10-20 min at a frequency of 20-30kHz, with ultrasonic treatment for 5 s and a 5 s interval.

[0025] Those skilled in the art can selectively adjust the above scheme to obtain the intracellular contents of the corresponding strain.

[0026] In another aspect, the present invention provides the use of the aforementioned probiotic composition in the preparation of anti-allergy products.

[0027] The aforementioned anti-allergy measures can prevent or alleviate allergy symptoms.

[0028] The product may be a health supplement or a medicine.

[0029] The health products may also include other excipients.

[0030] The medicine may also include other pharmaceutically acceptable excipients.

[0031] The pharmaceutically acceptable excipients include, but are not limited to, excipients, buffers, emulsifiers, stabilizers, diluents, binders, preservatives, and lubricants.

[0032] Specifically, the excipient is selected from at least one of microcrystalline cellulose, lactose, pregelatinized starch, cyclodextrin, carboxymethyl cellulose, and mannitol.

[0033] Specifically, the buffer is selected from at least one of sodium dihydrogen phosphate, sodium bicarbonate, ammonium bicarbonate, sodium acetate, citrate, histidine, and succinate.

[0034] Specifically, the emulsifier is selected from at least one of magnesium stearate, zinc stearate, calcium stearate, glyceryl stearate, sorbitan isostearate, sorbitan oleate, glyceryl oleate, and polyglycerol 3-polyricinoleate.

[0035] Specifically, the stabilizer is selected from at least one of acacia gum, agar, alginic acid, cellulose ether, and carboxymethyl chitosan.

[0036] Specifically, the diluent is selected from at least one of erythritol, mannitol, sorbitol, xylitol, lactose, sucrose, corn starch, potato starch, calcium phosphate, calcium citrate, and crystalline cellulose.

[0037] Specifically, the adhesive is selected from at least one of ethanol, starch paste, syrup, hydroxypropyl methylcellulose, sodium carboxymethyl cellulose, sodium alginate, and polyvinylpyrrolidone.

[0038] Specifically, the preservative is selected from at least one of methylparaben, propylparaben, methylparaben, ethylparaben, propylparaben, chlorobutanol, thimerosal, mercuric oxycyanide, phenoxyethanol, chlorhexidine, benzoic acid, sodium benzoate, chlorocresol, benzalkonium bromide, benzalkonium chloride, and ethylparaben.

[0039] Specifically, the lubricant is selected from at least one of magnesium stearate, stearic acid, sodium chloride, sodium oleate, sodium lauryl sulfate, and poloxamer.

[0040] The beneficial effects of this invention are:

[0041] This invention optimizes the application form of Bacillus coagulans (bacterial powder + post-biotic) and combines it with other raw materials (fructooligosaccharides, lactitol, acerola cherry powder and stachyose) to prepare a composition that can effectively relieve allergies. Compared with other strains in the prior art (such as Lactobacillus rhamnosus LGG), it has a more significant effect, and as a whole, it has a better effect on animal models. Detailed Implementation

[0042] The present invention will be further described in detail below with reference to specific embodiments. The following embodiments are not intended to limit the present invention, but only to illustrate the present invention. Unless otherwise specified, the experimental methods used in the following embodiments are generally performed under conventional conditions. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available.

[0043] In the following embodiments, some of the raw materials used are from the following sources:

[0044] The fructooligosaccharides were purchased from Quantum Hi-Tech (Guangdong) Biotechnology Co., Ltd.

[0045] Lactitol was purchased from Shandong Lvjian Biotechnology Co., Ltd.

[0046] Acerola cherry powder was purchased from Tianjin Zhenruoguo Food Industry Co., Ltd.

[0047] Stachyose was purchased from Anhui Zhongxinkang Pharmaceutical Co., Ltd.

[0048] Example 1: A probiotic composition for improving allergies

[0049] The probiotic composition formula (10g) in this embodiment is as follows:

[0050] Bacillus coagulans BC01 powder (live bacteria content 10) 9 -10 10 CFU / g, in this example, is 5 × 10⁻⁶. 9 CFU / g): 2g;

[0051] Bacillus coagulans BC01 postbiotic: 1g; fructooligosaccharide 1g, lactitol 0.5g, acerola cherry powder 1g and stachyose 2g.

[0052] Bacillus coagulans BC01, with accession number CCTCC NO: M2017813, is a prior art strain, with specific information disclosed in patent applications CN202310194904.1 and CN202110260653.3.

[0053] The preparation method of Bacillus coagulans BC01 bacterial powder is as follows:

[0054] 1%-5% of Bacillus coagulans BC01 was inoculated into fermentation medium and cultured for 24 hours to obtain fermentation broth. The broth was then centrifuged at 10,000 rpm for 20 minutes to collect the bacterial cells, which were then freeze-dried to obtain bacterial powder.

[0055] The preparation method of Bacillus coagulans BC01 postbiotic is as follows:

[0056] 1%-5% of Bacillus coagulans BC01 was inoculated into fermentation medium and cultured for 24 hours to obtain fermentation broth. The bacterial cells were collected, washed with sterile PBS, resuspended, and the bacterial concentration was adjusted to 1×10⁻⁶. 9 -10×10 9 CFU / mL (5×10⁻⁶ in this example) 9The bacterial suspension was subjected to high temperature and high pressure treatment to obtain inactivated bacteria (CFU / mL); an ice-water mixture was prepared as the ultrasonic medium and treated at 500-600W power (550W in this example) for 15 minutes at a frequency of 25kHz. The ultrasonic probe was placed 4cm below the water surface for 5s working and 5s intermittent ultrasonic treatment to obtain post-genetic agents.

[0057] Mix according to the formula, then dispense into smaller portions and store away from light.

[0058] Example 2

[0059] Referring to Example 1, the difference lies in the amount of each component used, as detailed below:

[0060] Bacillus coagulans BC01 bacterial powder (live bacteria content 5×10⁻⁶) 9 CFU / g): 1.5g; Bacillus coagulans BC01 postbiotic: 1.5g; fructooligosaccharides 1g, lactitol 1g, acerola cherry powder 1.5g and stachyose 1g.

[0061] Example 3

[0062] Referring to Example 1, the difference lies in the amount of each component used, as detailed below:

[0063] Bacillus coagulans BC01 bacterial powder (live bacteria content 5×10⁻⁶) 9 CFU / g): 2.5g; Bacillus coagulans BC01 postbiotic: 0.5g; fructooligosaccharides 2g, lactitol 1g, acerola cherry powder 0.5g and stachyose 1g.

[0064] Comparative Example 1

[0065] Referring to Example 1, the difference is that lactitol is replaced with xylitol.

[0066] Comparative Example 2

[0067] Referring to Example 1, the difference is that the Bacillus coagulans BC01 postbiotic is replaced with Bacillus coagulans BC01 bacterial powder.

[0068] Comparative Example 3

[0069] Referring to Example 1, the difference is that the Bacillus coagulans BC01 powder is replaced with Bacillus coagulans BC01 postbiotic.

[0070] Comparative Example 4

[0071] The composition (10g) is formulated as follows: Bacillus coagulans BC01 bacterial powder (live bacteria content 5×10⁻⁶). 9 CFU / g): 2g; Bacillus coagulans BC01 postbiotic: 1g. When using, the component concentrations are the same as in Example 1.

[0072] Comparative Example 5

[0073] The composition (10g) is formulated as follows: 1g fructooligosaccharide, 0.5g lactitol, 1g acerola cherry powder, and 2g stachyose. When using, the concentration of the components is the same as in Example 1.

[0074] Comparative Example 6

[0075] Referring to Example 1, the difference is that Bacillus coagulans is replaced with Lactobacillus rhamnosus LGG (ATCC 53103, type strain), which is a strain disclosed in the prior art.

[0076] Effect Experiment Example

[0077] SPF-grade male Brown-Norway (BN) rats, weighing (200±10) g. They were acclimatized for 7 days with a standard diet and free access to water. Ambient temperature: 24±1℃, humidity: 55±10%.

[0078] The allergic reaction model was constructed using ovalbumin (OVA) injection. OVA was purchased from Sigma, catalog number A5503, with a purity >98%, and was dissolved in 0.1M PBS at 1% (g / L). Construction method: On days 1, 7, and 14 after the start of the experiment, 1 mL of OVA solution was injected intraperitoneally. On day 21, 2 mL of OVA solution was injected intraperitoneally. Scoring was performed 7 days after the last intraperitoneal OVA injection, blood was collected from the tail vein, and the animals were euthanized.

[0079] Eleven groups were set up, with seven animals in each group. The samples administered by gavage were 1g of the composition prepared for each example or comparative example dissolved in 5mL of physiological saline. The specific treatment methods for each group are as follows:

[0080] Blank control group; 1 mL of 0.1 M PBS was injected intraperitoneally on days 1, 7, 14 and 21 after the start of the experiment, and 5 mL of physiological saline was administered by gavage daily.

[0081] Allergic reaction model group: The model was constructed according to the aforementioned model construction method, and 5 mL of physiological saline was administered by gavage daily;

[0082] Example 1: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Example 1 was administered by gavage daily;

[0083] Example 2: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Example 2 was administered by gavage daily;

[0084] Example 3: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Example 3 was administered by gavage daily;

[0085] Comparative Example 1: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Comparative Example 1 was administered by gavage daily;

[0086] Comparative Example 2: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Comparative Example 2 was administered by gavage daily;

[0087] Comparative Example 3: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Comparative Example 3 was administered by gavage daily;

[0088] Comparative Example 4: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Comparative Example 4 was administered by gavage daily;

[0089] Comparative Example 5: A model was constructed according to the aforementioned model construction method, and 5 mL of the composition of Comparative Example 5 was administered by gavage daily;

[0090] Comparative Example 6: A model was constructed using the aforementioned model construction method, and 5 mL of the composition of Comparative Example 6 was administered by gavage daily.

[0091] Testing indicators:

[0092] (1) Allergy Symptom Score

[0093] Allergic symptoms were scored before mice were sacrificed, following common scoring methods in existing technologies. Specific scoring criteria can be found at:

[0094] 0 points: No symptoms; 1 point: Scratching face; 2 points: Facial redness and swelling, rapid breathing, lethargy; 3 points: Tearing, red rash around the mouth and tail, asthma, difficulty breathing; 4 points: Inactivity, muscle contraction, spasm; 5 points: Death, shock.

[0095] The average value was calculated after the scores of mice in each group were tallied.

[0096] (2) Detection of ovalbumin-specific antibodies

[0097] Commercial ELISA kits were used to detect OVA-IgE (UKW-20264) and OVA-IgG (QY-XS2391-1), and the detection steps were performed according to the instructions.

[0098] (3) Serum inflammatory factors

[0099] All serum inflammatory factor detection kits were purchased from ELISA, with catalog numbers as follows: IL-17A Detection Kit ml037365; IL-4 Detection Kit ml102825; IFN-γ Detection Kit ml102842; TNF-α Detection Kit ml002859; and Histamine (HIS) Detection Kit ml002986. The detection procedures were performed according to the instructions.

[0100] Test results:

[0101] (1) The results of the allergy symptom score are shown in Table 1:

[0102] Table 1

[0103] Group Allergy Symptom Score (average ± STD) Blank control 0.0±0.0 Model group 3.8±0.4 Example 1 1.1±0.2 Example 2 1.5±0.3 Example 3 1.3±0.1 Comparative Example 1 1.8±0.0 Comparative Example 2 2.9±0.2 Comparative Example 3 3.2±0.5 Comparative Example 4 2.1±0.3 Comparative Example 5 3.6±0.4 Comparative Example 6 2.4±0.6

[0104] As shown in Table 1, the overall allergy symptom scores of the model group were relatively high, indicating successful model construction. Examples 1-3 showed some effect in alleviating allergy symptoms. Compared to Comparative Examples 1 and 6, it can be seen that the components in the overall formula of this invention have a better overall effect; compared to Comparative Examples 2 and 3, it can be seen that the combined effect of the bacterial powder and post-biotic is superior; compared to Comparative Examples 4 and 5, it can be seen that the overall effect between the bacterial component and other components is significant.

[0105] (2) The results of ovalbumin-specific antibody detection are shown in Table 2:

[0106] Table 2

[0107]

[0108]

[0109] Both OVA-IgE and OVA-IgG are OVA-specific antibodies, and their elevated levels can, to some extent, reflect allergic reactions in animal models. The levels of OVA-IgE and OVA-IgG in the model group were significantly higher than in the control group, indicating successful modeling. The compositions of Examples 1-3 and Comparative Examples 1-4 and 6 all played a role in the allergic reaction of the model, while the composition of Comparative Example 5 did not show a desensitizing effect. Furthermore, the effects of Examples 1-3 were significantly better than those of the comparative examples.

[0110] (3) The results of serum inflammatory factor detection are shown in Table 3:

[0111] Table 3

[0112]

[0113] IL-17A, IL-4, IFN-γ, TNF-α, and HIS are serum inflammatory factors associated with allergic reactions, and their levels were significantly increased in the model group. After gavage administration of the compositions of Examples 1-3, the serum inflammatory factors in the animal models decreased significantly, especially IL-17A and TNF-α, which showed a significant improvement compared to the control group. Other factors also demonstrated the ability to control allergic reactions to some extent.

[0114] The above embodiments are only used to illustrate the present invention and are not intended to limit the present invention. Any conventional changes, substitutions and optimizations made by those skilled in the art based on the above technical solutions are within the protection scope of the present invention.

Claims

1. A probiotic composition for improving allergy, characterized by, It comprises by weight parts: Bacillus coagulans BC01 bacteria powder 2 parts, Bacillus coagulans BC01 postbiotic 1 part, fructooligosaccharide 1 part, lactitol 0.5 part, needle cherry powder 1 part and stachyose 2 parts. Bacillus coagulans BC01 powder, Bacillus coagulans BC01 probiotic, fructo-oligosaccharide, lactitol, pin cherry powder and stachyose, the use ratio of the Bacillus coagulans BC01 powder and the Bacillus coagulans BC01 probiotic is 1-5:1, the content of the probiotic in the probiotic composition is 10 9 -10 10 CFU / g, by weight parts, including: 1.5-2.5 parts of Bacillus coagulans BC01 powder, 0.5-1.5 parts of Bacillus coagulans BC01 probiotic, 1-2 parts of fructo-oligosaccharide, 0.5-1 part of lactitol, 0.5-1.5 parts of pin cherry powder and 1-2 parts of stachyose.

2. The probiotic composition according to claim 1, characterized in that, The preparation method of the Bacillus coagulans BC01 postbiotic is ultrasonic crushing.

3. The probiotic composition according to claim 2, characterized in that, The content of the probiotic bacteria in the Bacillus coagulans BC01 bacterial powder is 5 x 10 9 CFU / g.

4. The probiotic composition according to claim 3, characterized in that, The preparation method of the Bacillus coagulans BC01 postbiotic is:

5. The probiotic composition according to claim 4, characterized in that, It comprises: The fermentation broth was obtained by inoculating 1%-5% Bacillus coagulans BC01 in the culture medium, collecting the bacterial cells, and resuspending the bacterial cells in PBS to a concentration of 1×10 9 -10×10 9 CFU / mL, inactivated; the inactivated bacteria were treated at a power of 500-600 W for 10-20 min at a frequency of 20-30 kHz, and the ultrasonic treatment was performed for 5 s and then stopped for 5 s.

6. A method of preparing a probiotic composition for improving allergy, characterized by, (1) preparing Bacillus coagulans BC01 bacteria powder and Bacillus coagulans BC01 postbiotic; (2) compounding according to the formula of the probiotic composition of any one of claims 1-3.

7. Use of the probiotic composition of any one of claims 1-5 in the preparation of an anti-allergic product. ​

Images