Compound probiotic formula, preparation and application thereof

Abstract

The invention relates to a compound probiotic formula, a preparation and application of the compound probiotic formula. The compound probiotic formula is prepared from bifidobacterium animalis HN019, bifidobacterium animalis Bb-12, bifidobacterium breve M-16V, lactobacillus rhamnosus HN001 and lactobacillus acidophilus NCFM. The ratio of the viable count of the bifidobacterium animalis HN019 to the viable count of the bifidobacterium animalis Bb-12 to the viable count of the bifidobacterium breve M-16V to the viable count of the lactobacillus rhamnosus HN001 to the viable count of the lactobacillus acidophilus NCFM is 2.5 to 1.2 to 1.3 to 2.5 to 2.5. The traditional Chinese medicine composition has the advantages of helping to establish intestinal flora balance, improving immunity, preventing infectious diseases, regulating immune balance, preventing and relieving eczema and allergy, bidirectionally regulating diarrhea and constipation, relieving gastrointestinal function discomfort symptoms, improving poor appetite, anorexia and dyspepsia, improving children crying and screaming at night and poor sleep, improving children growth and development retardation, and improving children immunity. Social emotion behaviors of children are improved, and attention deficits are improved; the probiotic composition can coexist and symbiotic, induces an organism to establish intestinal flora balance, relieves food allergy symptoms, and improves social emotion problems caused by food allergy.

Description

Technical Field

[0001] The present invention relates to the technical field of probiotic formula, and more specifically to a composite probiotic formula, preparation and application thereof. Background Art

[0002] Probiotics are living microorganisms that, when given in sufficient amounts, can regulate the host's immune system and benefit the human body. The main physiological function of probiotics is to maintain the balance of intestinal flora and regulate the immune system, thereby playing an anti-inflammatory role and alleviating chronic diseases. More and more studies have shown that the number and type of intestinal flora can interfere with the occurrence of food allergies. Therefore, the preventive and therapeutic effects of probiotics on food allergies are increasingly gaining attention.

[0003] At the same time, as research progresses, it is found that food allergies can cause a variety of behavioral development problems in children, such as attention deficit, impulsiveness, social avoidance, anxiety, depression, etc. However, since the probiotics on the market only target the effects of intestinal damage and immune imbalance caused by food allergies, and have not been further tested on the role of probiotics in neurobehavioral development problems caused by food allergies, there is not enough evidence for the role of probiotics in this regard.

[0004] The preceding description is intended to provide general background information and does not necessarily constitute prior art. Summary of the invention

[0005] The purpose of the present invention is to provide a composite probiotic formula, the composite probiotic formula probiotic combination can coexist and coexist, induce the body to establish a balance of intestinal flora, relieve food allergy symptoms, and improve social emotional problems, attention deficit and other behavioral changes caused by food allergies.

[0006] The invention provides a composite probiotic formula, which comprises animal bifidobacterium HN019, animal bifidobacterium Bb-12, bifidobacterium breve M-16V, rhamnosaurus HN001 and lactobacillus acidophilus NCFM; the ratio of the live bacteria counts of the animal bifidobacterium HN019, the animal bifidobacterium Bb-12, the bifidobacterium breve M-16V, the rhamnosaurus HN001 and the lactobacillus acidophilus NCFM is 2.5:1.2:1.3:2.5:2.5.

[0007] Furthermore, each 2g of probiotic freeze-dried powder contains:

[0008] Bifidobacterium animalis HN019: 2.5 billion;

[0009] Bifidobacterium animalis Bb-12: 1.2 billion;

[0010] Bifidobacterium breve M-16V: 1.3 billion;

[0011] Lactobacillus rhamnosus HN001: 2.5 billion;

[0012] Lactobacillus acidophilus NCFM: 2.5 billion.

[0013] The present invention also provides a composite probiotic preparation, which is prepared from the composite probiotic formula.

[0014] The present invention also provides a composite probiotic application, which includes the application of the composite probiotic formula in improving intestinal function and / or abnormal brain development caused by allergies.

[0015] The composite probiotic formula provided by the present invention, wherein the probiotics are selected from 500-1000 kinds of probiotics in the intestine, the probiotics that have been proven to be beneficial to human life and health, and the probiotics that are helpful for the prevention and treatment of food allergies are selected, and the selected probiotics can coexist symbiotically, and the scientific combination, feeding and supplementation, and two-pronged approach have more significant effects on comprehensively regulating the intestinal ecological balance and increasing the value of the intestinal native flora; on the other hand, the probiotic formula of the present invention can help establish the balance of intestinal flora, and the application indications for all groups of people (including children) are: improving immunity, preventing infectious diseases, regulating immune balance, preventing and relieving eczema and allergies, bidirectionally regulating diarrhea and constipation, relieving gastrointestinal dysfunction symptoms, improving poor appetite, anorexia, indigestion, improving children's crying and poor sleep at night, improving children's growth retardation, improving children's social emotional behavior, and improving attention deficit; on the other hand, the probiotic combination of the present invention can coexist symbiotically, induce the body to establish intestinal flora balance, relieve food allergy symptoms, and improve social emotional problems, attention deficit and other behavioral changes caused by food allergies. DETAILED DESCRIPTION

[0016] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

[0017] The terms "first", "second", "third", "fourth" and the like in the description and claims of the present invention are used to distinguish similar objects and are not necessarily used to describe a particular order or sequence.

[0018] Example 1

[0019] An embodiment of the present invention provides a composite probiotic formula, which includes animal Bifidobacterium HN019, animal Bifidobacterium Bb-12, Bifidobacterium breve M-16V, Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus NCFM; the ratio of the number of live bacteria of the animal Bifidobacterium HN019, the animal Bifidobacterium Bb-12, the Bifidobacterium breve M-16V, the Lactobacillus rhamnosus HN001 and the Lactobacillus acidophilus NCFM is 2.5:1.2:1.3:2.5:2.5.

[0020] It should be noted that the composite probiotic formula provided by the present invention, wherein the probiotics are selected from 500-1000 kinds of probiotics in the intestine, the probiotics that have been proven to be beneficial to human life and health, and the probiotics that are helpful for the prevention and treatment of food allergies are selected, and the above-mentioned probiotics can coexist symbiotically, and the scientific combination, the combination of feeding and supplementation, and the two-pronged approach have more significant effects, comprehensively regulate the intestinal ecological balance, and increase the value of the intestinal native flora;

[0021] On the other hand, the probiotic formula of the present invention can help establish a balance of intestinal flora, and the indications for use by all groups of people (including children) include: improving immunity, preventing infectious diseases, regulating immune balance, preventing and relieving eczema and allergies, bidirectionally regulating diarrhea and constipation, relieving symptoms of gastrointestinal discomfort, improving poor appetite, anorexia, and indigestion, improving children's nighttime crying and poor sleep, improving children's growth retardation, improving children's social emotional behavior, and improving attention deficit;

[0022] The probiotic combination of the present invention can coexist and coexist, inducing the body to establish a balance of intestinal flora, relieving food allergy symptoms, and improving behavioral changes such as social emotional problems and attention deficit caused by food allergies.

[0023] Furthermore, each 2g of probiotic freeze-dried powder contains:

[0024] Bifidobacterium animalis HN019: 2.5 billion;

[0025] Bifidobacterium animalis Bb-12: 1.2 billion;

[0026] Bifidobacterium breve M-16V: 1.3 billion;

[0027] Lactobacillus rhamnosus HN001: 2.5 billion;

[0028] Lactobacillus acidophilus NCFM: 2.5 billion.

[0029] The five-in-one probiotics in the present invention can regulate intestinal microecology and reduce the proportion of pathogenic bacteria; prevent the incidence of antibiotic-related diarrhea; increase the content of fecal short-chain fatty acids (acetic acid), and increased acetic acid promotes defecation; reduce total IgE and HDM-IgG1, reduce proinflammatory cytokines related to Th2, and reduce inflammation; increase the proportion of Treg and Th17 cells to effectively relieve allergic symptoms; significantly change the levels of TGF-β1, TGF-β1 and IgA; clinical trials for the treatment of diarrhea have shown that it can reduce the incidence of diarrhea; clinical trials for the treatment of constipation have shown that it can reduce the pain and abdominal pain caused by constipation defecation and improve constipation.

[0030] The present invention also provides a composite probiotic preparation, which is prepared from the composite probiotic formula.

[0031] It should be noted that the strains in the probiotic formula of the present invention are directly made into probiotic preparations for consumption. The transportation and storage of the finished product are the same as those of general probiotics, and the temperature can be kept below 25°C without special requirements. The finished product can also be added to other products, including food, medicine or health food.

[0032] The present invention also provides a composite probiotic application, which includes the application of the composite probiotic formula in improving intestinal function and / or abnormal brain development caused by allergies.

[0033] Example 2

[0034] In this example, male C57BL / 6J mice were studied using the probiotic freeze-dried powder obtained by the composite probiotic formula of the present invention.

[0035] The specific experimental steps include the following:

[0036] S1: Male C57BL / 6J mice were randomly divided into three groups: normal control group, ovalbumin (OVA) food allergy group, and probiotic intervention group. The probiotic intervention group was given probiotics by gavage for 30 consecutive days based on OVA food allergy.

[0037] S2: Mouse neurobehavior was assessed using three-box social interaction, elevated maze, open field, grooming test, and non-selective and non-sustained attention tests;

[0038] S3: Enzyme-linked immunosorbent assay, immunohistochemistry, 16s DNA sequencing technology, and non-targeted metabolomics analysis technology were used to evaluate the improvement effects of probiotic intervention on neuroinflammation, intestinal / brain pathology, intestinal flora and metabolites in food-allergic mice.

[0039] The results of the study showed that after taking the probiotic formula of the present invention, the intestinal damage of food allergic mice was significantly improved, and the neurobehavioral abnormalities were restored to a certain extent.

[0040] In addition to being used to prevent and treat intestinal damage caused by food allergies, the above-mentioned probiotic formula can also be used to treat neurobehavioral changes caused by food allergies.

[0041] ① Improvement of food allergy and immune function: Compared with the control group, the allergic clinical symptom score of the OVA food allergy group increased significantly (t=6.79, P<0.001), and probiotic intervention could significantly alleviate allergic symptoms (t=2.63, P<0.05). The serum OVA-IgE, OVA-IgG1 and histamine levels of the OVA food allergy group were significantly higher than those of the control group (tOVA-IgE=8.78, P<0.01; tOVA-IgG1=13.79, P<0.01; thistamine=4.80, P<0.01), while probiotic intervention could reduce the levels of the three (tOVA-IgE=7.55, P<0.01; tOVA-IgG1=10.71, P<0.01; thistamine=3.14, P<0.01).

[0042] ②Improving gastrointestinal function damage: The OVA food allergy group had severe intestinal villi damage, while the probiotic intervention group significantly improved the pathological damage;

[0043] ③ Improve social emotions and enhance attention: Compared with the control group, the social index (t=2.32, P<0.05) and social novelty preference index (t=2.22, P<0.05) of OVA food allergic mice were significantly decreased, repetitive grooming behavior increased (t=3.48, P<0.01), the time of exploring the open arm was reduced (t=4.10, P<0.01), and the attention level decreased (t=3.12, P<0.01). Probiotic intervention can improve social behavior (t=2.69, P<0.05), social novelty preference (t=2.43, P<0.05), stereotyped behavior (t=2.18, P<0.05), anxiety (t=2.28, P<0.05) and attention (t=3.09, P<0.01);

[0044] ④Improvement of brain neuron damage: Immunohistochemistry results showed that compared with the control group, the OVA food allergy group had abnormal activation of prefrontal microglia (t=2.53, P<0.05) and a decrease in the number of Nissl body-positive neurons (t=2.67, P<0.05), while probiotic intervention could inhibit microglia activation (t=2.78, P<0.05) and alleviate neuron damage (t=3.81, P<0.01);

[0045] ⑤ Improve the health of intestinal flora: In the 16s DNA sequencing of intestinal feces, compared with the control group, the abundance of Lactobacillaceae and Bifidobacteriaceae decreased in the OVA food allergy group (P<0.05); after probiotic intervention, the abundance of Lactobacillaceae and Bifidobacteriaceae increased (P<0.05), and α diversity improved;

[0046] ⑥ Improvement of intestinal flora-related metabolites and brain function in serum: Serum metabolomics results showed that compared with the control group, the differential metabolites in the serum of OVA food-allergic mice were mainly enriched in the tricarboxylic acid cycle, among which the level of succinate related to inflammatory response was increased. After probiotic intervention, the levels of key metabolites of the tricarboxylic acid cycle pathway in the serum of mice were reversed, and the succinate level was reduced; the correlation analysis of the serum succinate level and behavioral indicators of mice showed that the succinate level was significantly negatively correlated with the social index (R2=0.352, P<0.05), the length of stay in the open arm of the elevated maze (R2=0.350, P<0.05), and the attention level (R2=0.303, P<0.05), and was positively correlated with the length of grooming time (R2=0.413, P<0.05).

[0047] The above-mentioned probiotics can improve the intestinal flora disorders caused by food allergies, alleviate serum metabolite abnormalities, improve neuroinflammation and neuronal damage, and thus improve neurobehavioral changes in food allergic mice.

[0048] Based on the above description, it can be seen that the advantages of the present invention are:

[0049] 1. The composite probiotic formula provided by the present invention, wherein the probiotics are selected from 500-1000 kinds of probiotics in the intestine, the probiotics that have been proven to be beneficial to human life and health, and the probiotics that are helpful for the prevention and treatment of food allergies are selected, and the above-mentioned probiotics can coexist symbiotically, and the scientific combination, feeding and supplementation, and two-pronged approach have more significant effects, comprehensively regulate the intestinal ecological balance, and increase the value of the intestinal native flora;

[0050] 2. The compound probiotic formula provided by the present invention can help establish the balance of intestinal flora. The indications for application of the compound probiotic formula for all groups of people (including children) are: improving immunity, preventing infectious diseases, regulating immune balance, preventing and relieving eczema and allergies, bidirectionally regulating diarrhea and constipation, relieving gastrointestinal discomfort symptoms, improving poor appetite, anorexia, indigestion, improving children's crying and poor sleep at night, improving children's growth retardation, improving children's social emotional behavior, and improving attention deficit;

[0051] 3. The composite probiotic formula provided by the present invention and the probiotic combination can coexist and coexist, inducing the body to establish a balance of intestinal flora, relieving food allergy symptoms, and improving social emotional problems, attention deficit and other behavioral improvements caused by food allergies.

[0052] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A composite probiotic formula, characterized in that: The composite probiotic formula includes Bifidobacterium animalis HN019, Bifidobacterium animalis Bb-12, Bifidobacterium breve M-16V, Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus NCFM; The ratio of the number of live bacteria of the Bifidobacterium animalis HN019, the Bifidobacterium animalis Bb-12, the Bifidobacterium breve M-16V, the Lactobacillus rhamnosus HN001 and the Lactobacillus acidophilus NCFM is 2.5:1.2:1.3:2.5:2.

5.

2. The composite probiotic formula according to claim 1, characterized in that: Each 2g of probiotic freeze-dried powder contains: Bifidobacterium animalis HN019: 2.5 billion; Bifidobacterium animalis Bb-12: 1.2 billion; Bifidobacterium breve M-16V: 1.3 billion; Lactobacillus rhamnosus HN001: 2.5 billion; Lactobacillus acidophilus NCFM: 2.5 billion.

3. A composite probiotic preparation, characterized in that: The composite probiotic preparation is prepared from the composite probiotic formula according to any one of claims 1 to 2.

4. A composite probiotic application, characterized in that: The application of the composite probiotics includes the application of the composite probiotic formula described in any one of claims 1 to 2 in improving intestinal function and / or brain development abnormalities caused by allergies.