Enterococcus faecalis compositions and uses in treating intestinal diseases and modulating immunity

Abstract

The application discloses an Enterococcus faecalis composition and application thereof in treating intestinal diseases and regulating immunity, and belongs to the technical field of microorganisms. The technical problem to be solved is to provide a composition with improved effect in treating intestinal diseases and regulating immunity and reduced dosage. The technical solution is as follows: the composition comprises Enterococcus faecalis and lactoferrin, wherein the Enterococcus faecalis is preserved in the Guangdong Microbial Culture Collection Center on October 13, 2022, and the preservation number is GDMCC NO. 62869, the weight ratio of the lactoferrin and the Enterococcus faecalis is 10-600:1, and the bacterial amount of the Enterococcus faecalis is 1 billion CFU / g-10,000 billion CFU / g.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology. More specifically, it relates to Enterococcus faecalis compositions and their use in treating intestinal diseases and regulating immunity. Background Technology

[0002] Enterococcus faecalis is a normal commensal flora in the human gut, and its core functions include: (1) Maintain the balance of intestinal flora, inhibit the excessive proliferation of harmful bacteria such as Escherichia coli and Salmonella, and stabilize the intestinal microecological environment; (2) Assist in nutrient metabolism: participate in the decomposition of complex carbohydrates in food, synthesize B vitamins, metabolize lactic acid to reduce intestinal pH, promote the conversion of calcium into calcium lactate, and improve nutrient absorption efficiency; (3) Regulate immune function: as a non-specific immune regulatory factor, it gently stimulates the development of intestinal lymphoid tissue, enhances macrophage activity, and improves the body's overall resistance; (4) Improve digestive efficiency: it can soften fiber, decompose some proteins into small molecule peptides, and improve the digestion and utilization rate of nutrients.

[0003] Lactoferrin is an iron-binding glycoprotein in the transferrin family. It is a core immune-active protein widely found in mammalian milk and secretions, possessing a variety of physiological functions such as broad-spectrum antibacterial, antiviral, immune regulation, and iron absorption promotion.

[0004] The patent, published in China (CN117511773A) on February 6, 2024, discloses a strain of *Enterococcus faecalis*, a compound microbial agent, and their applications, relating to the field of microbiology. Specifically, the *Enterococcus faecalis* (… Enterococcus faecalis The *Enterococcus faecalis* and its compound preparation were deposited at the Guangdong Provincial Center for Microbial Culture Collection on October 13, 2022, with accession number GDMCC NO.62869. These *Enterococcus faecalis* and their compound preparations are effective in treating feline intestinal disorders and feline inflammatory bowel disease, and can be used in the preparation of related drugs.

[0005] This patent, published in the United States (US20130273015A1) on October 17, 2013, discloses nutritional compositions specifically designed to meet the needs of infants and young children at least two years of age. Specifically, the invention provides a set of nutritional compositions for infants and young children, each with a different probiotic content. Examples of suitable microorganisms include Enterococcus faecalis, and in a preferred embodiment, any composition forming part of this set of nutritional compositions may contain lactoferrin. Effects include treating or preventing diarrhea, treating or preventing intestinal discomfort, and promoting the maturation of the immune system.

[0006] The study "The Effects of Lactoferrin on the Intestinal Microbiota of Mice" (Wei Xiuxiu, Chen Qingsen et al., Food Industry Technology) describes how mice were randomly divided into five groups after one week of feeding: a control group, a placebo group, and low, medium, and high lactoferrin groups. The control group received a normal diet, the placebo group was administered 0.2 mL of physiological saline by gavage, and the experimental groups were administered different doses of lactoferrin solution by gavage. These groups were fed continuously. The fecal microbiota of the mice was measured using traditional culture techniques, and the changes in the intestinal microbiota were analyzed through data analysis. The results showed that the number of lactobacilli and bifidobacteria in the mouse intestines significantly increased after lactoferrin administration, while the number of enterobacteria and enterococci significantly decreased. This indicates that a certain dose of lactoferrin has a significant promoting effect on the growth of beneficial bacteria and resident symbiotic bacteria in the intestines, while having a certain inhibitory effect on enterobacteria, enterococci, and other bacteria.

[0007] The prior art represented by the aforementioned literature has at least the following unresolved technical problems: (1) The patent document with publication number CN117511773A provides a strain of Enterococcus faecalis. To improve immunity and relieve diarrhea, it is necessary to further explore the formulation composition in order to reduce the dosage and improve the effect.

[0008] (2) The combination of probiotics and lactoferrin has been disclosed, but there are differences between different strains and the synergistic mechanism when combined with lactoferrin is different. Further selection is still needed to achieve synergistic effect through specific strains. Summary of the Invention

[0009] The purpose of this invention is to provide: A composition having therapeutic effects on intestinal diseases and immune regulation, and related technologies thereof, to address technical issues such as improving the efficacy of treating intestinal diseases and regulating immunity, reducing dosage, or combinations thereof.

[0010] Terminology Explanation: Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this subject matter pertains. Unless otherwise stated, all patents, patent inventions, and disclosures cited throughout this document are incorporated herein by reference in their entirety. Where multiple definitions exist for terms herein, the definitions provided in this chapter shall prevail.

[0011] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0012] Unless otherwise stated, conventional methods within the scope of the art, such as heating, filtration, and volume adjustment, shall be used.

[0013] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0014] The term “therapeutic effective dose” as used in this article refers to a dose that is sufficient to achieve the desired therapeutic outcome or to affect undesirable symptoms, but is generally insufficient to cause adverse side effects.

[0015] The term “safe effective dose” as used in this article refers to a dose sufficient to produce the desired effect but low enough to avoid serious side effects.

[0016] As used herein, the terms “subject” or “patient” may refer to a vertebrate organism, such as a mammal (e.g., a human). “Subject” may also refer to a cell, cell group, tissue, organ, or organism, preferably a mammal.

[0017] As used herein, the term "treatment" generally refers to achieving the desired pharmacological and / or physiological effect. An effect may be preventative, but not necessarily preventative, in relation to the prevention or partial prevention of a disease, symptom, or condition. An effect may be therapeutic in relation to the partial or complete cure of a disease, condition, symptom, or adverse reaction attributable to that disease, condition, or condition. Specifically, the term "treatment" can include any treatment of a subject (particularly mammals) for a relevant symptom or disease, and may include any one or more of the following: (a) prevention of the disease in subjects who may be susceptible to the disease but have not yet been diagnosed with it; (b) suppression of the disease, i.e., prevention of its development; and (c) relief of the disease, i.e., reduction or improvement of the disease and / or its symptoms or condition. The term "treatment" can refer to therapeutic treatment alone, preventative treatment alone, or both therapeutic and preventative treatment. Mammals requiring treatment (subjects requiring treatment) can include mammals that already have a symptom and / or mammals that require prevention of a symptom.

[0018] In a first aspect, the present invention provides: an Enterococcus faecalis composition comprising: Enterococcus faecalis ( Enterococcus faecalisThe bacteria are lactoferrin and enterococci, wherein the Enterococcus faecalis was deposited at the Guangdong Provincial Center for Microbial Culture Collection on October 13, 2022, with accession number GDMCC NO. 62869, the weight ratio of lactoferrin to Enterococcus faecalis is 10-600:1, and the bacterial count of Enterococcus faecalis is 1 billion CFU / g-1 trillion CFU / g.

[0019] Among them are technical features: specific strains of Enterococcus faecalis, lactoferrin, and the weight ratio of Enterococcus faecalis and lactoferrin.

[0020] In this invention, Enterococcus faecalis is a specific strain. This invention has found that the combination of a specific strain of Enterococcus faecalis and lactoferrin has significant synergistic effects, and can achieve good therapeutic effects on diarrhea and immune regulation at lower doses.

[0021] The lactoferrin may be selected from bovine lactoferrin, human lactoferrin, porcine lactoferrin, sheep lactoferrin, recombinant human lactoferrin, or a combination thereof.

[0022] Furthermore, the weight ratio of lactoferrin to Enterococcus faecalis is 15-400:1.

[0023] Furthermore, the weight ratio of lactoferrin to Enterococcus faecalis is 15-250:1.

[0024] The bacterial count of Enterococcus faecalis was 800 billion CFU / g.

[0025] Furthermore, the Enterococcus faecalis is the bacterial cell of Enterococcus faecalis or its preparation.

[0026] Furthermore, the prepared product is selected from at least one of fermentation broth, fermentation broth precipitate, fermentation broth supernatant, live bacteria, inactivated bacteria, lyophilized powder, lysate, lysate, secondary metabolites, and exosomes.

[0027] Specifically, the fermentation broth is a mixed liquid system containing the bacteria themselves, intracellular and extracellular metabolites, unused culture medium components, and fermentation byproducts obtained by culturing Enterococcus faecalis in a culture medium under artificially controlled fermentation conditions.

[0028] Specifically, the sediment of Enterococcus faecalis fermentation broth is the solid phase component separated from the fermentation broth of the strain after treatment such as standing, centrifugation or filtration. It mainly includes live / dead cells of the strain, cell fragments and insoluble substances produced in the fermentation system.

[0029] Specifically, the supernatant of Enterococcus faecalis fermentation broth is a clear liquid phase component containing extracellular metabolites of the strain, soluble culture medium residues, and soluble fermentation byproducts obtained after the fermentation broth of the strain has been allowed to stand, centrifuged, or filtered to remove solid phase precipitates such as bacterial cells.

[0030] Specifically, live Enterococcus faecalis are bacteria with normal physiological activity that can carry out life activities such as metabolism and reproduction.

[0031] Specifically, inactivated Enterococcus faecalis refers to bacteria that have lost their metabolic and reproductive activities but whose overall structure has been basically preserved after being treated by physical, chemical or other means.

[0032] Specifically, Enterococcus faecalis freeze-dried powder refers to a solid powder that retains the original active components, obtained by removing moisture from liquid materials such as strain fermentation broth, fermentation supernatant, and bacterial suspension through a freeze-drying process.

[0033] Specifically, Enterococcus faecalis lysate refers to the mixture of intracellular substances and cell fragments formed after live or inactivated Enterococcus faecalis bacteria have had their cell walls broken by physical, chemical, or enzymatic methods, releasing intracellular substances.

[0034] Specifically, Enterococcus faecalis lysate refers to a mixed system containing intracellular active components and fragmented bacterial cells formed after the cell walls and cell membranes of live or inactivated Enterococcus faecalis are ruptured through physical, chemical, enzymatic, or biological lysation methods, releasing all intracellular substances.

[0035] Specifically, Enterococcus faecalis secondary metabolites refer to various compounds produced by microorganisms such as strains during their stable growth phase that are not essential for their own growth and reproduction and often possess specific biological activities such as anti-inflammatory and metabolic regulation.

[0036] Specifically, Enterococcus faecalis exosomes refer to extracellular vesicles encapsulated by nanoscale lipid bilayer membranes that are actively secreted by Enterococcus faecalis during its growth and metabolism or released after cell lysis.

[0037] Based on further solutions to the technical problems of the present invention, or simultaneous solutions to multiple technical problems, the preferred solution in the technical solution provided in the first aspect of the present invention includes: The weight ratio of lactoferrin to Enterococcus faecalis is 10-600:1; further, the weight ratio is 15-400:1; and even further, the weight ratio is 15-250:1. These preferred solutions, while addressing the technical problem of "improving the synergistic effect between components and enhancing the therapeutic effect on intestinal diseases and immune regulation," further strengthen the equivalent technical effects.

[0038] In a second aspect, the present invention provides an active formulation comprising the above-described composition.

[0039] As an alternative implementation, the active formulation also contains pharmaceutically acceptable excipients.

[0040] As an alternative implementation, the pharmaceutically acceptable excipients include any one or a combination of at least two of the following: carriers, diluents, excipients, fillers, binders, wetting agents, emulsifiers, cosolvents, surfactants, or buffers.

[0041] As an alternative implementation, the dosage form of the active preparation is selected from decoctions, ointments, pills, lozenges, tablets, granules, powders, capsules, patches, oral liquids, or syrups.

[0042] Suitable drug carriers can be selected for different dosage forms. For example, the drug carrier used can be solid, liquid, or gaseous. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0043] In preparing compositions for oral dosage forms, any convenient pharmaceutical medium can be used. For example, water, ethanol, oils, alcohols, flavoring agents, preservatives, coloring agents, etc., can be used to form oral liquid dosage forms, such as suspensions and solutions; while carriers, such as starch, sugars, microcrystalline cellulose, diluents, granulators, emulsifiers, lubricants, binders, and disintegrants, can be used to form oral solid dosage forms, such as powders, capsules, and tablets. Tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques.

[0044] Tablets containing the compositions of the present invention can be prepared by compression or molding, optionally using one or more excipients or adjuvants. The active ingredient can be compressed in a free-flowing form (e.g., powder or granules) in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, surfactant, or dispersant. Molded tablets can be molded in a suitable machine, i.e., a mixture of powdered compounds moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of active ingredient. For example, a formulation intended for oral administration to humans may contain about 0.5 mg to about 5 g of active pharmaceutical ingredient, mixed with a suitable and convenient carrier material, which may comprise about 5% to 95% of the total composition. Unit dosage forms typically contain about 1 mg to about 2 g of active ingredient, usually 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg, forming a mixture containing a softening or melting carrier, which is then cooled and shaped in a mold.

[0045] In addition to the carrier components mentioned above, the above pharmaceutical formulations may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. Furthermore, other excipients may be added, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, colorants, and flavoring agents.

[0046] Thirdly, the present invention provides the use of the above-described composition or the above-described active preparation in the preparation of products for the prevention and / or treatment of intestinal diseases.

[0047] The intestinal diseases mentioned include indigestion, malabsorption, vomiting, diarrhea, dysbiosis, irritable bowel syndrome, and inflammatory bowel diseases (such as ulcerative colitis and Crohn's disease).

[0048] Fourthly, the present invention provides the use of the above-described composition or the above-described active preparation in the preparation of products that regulate immunity.

[0049] Fifthly, the present invention provides a method for treating intestinal diseases or related symptoms thereof, comprising administering a subject or patient a therapeutically effective amount or a safe and effective amount of the above-described composition or the above-described active preparation.

[0050] Specifically, this method can be used in vivo or in vitro. The subject or patient can be a mammal, such as a human.

[0051] In the method described, the composition is a unit dosage form suitable for a single, precise dose. In some embodiments, the amount of the composition is in the range of about 0.001 mg / kg body weight / day to about 1000 mg / kg body weight / day. In some embodiments, the amount of the composition ranges from about 0.5 mg / kg body weight / day to about 50 mg / kg body weight / day. In some embodiments, the amount of the composition is about 0.001 g / day to about 7 g / day. In some embodiments, the amount of the composition is about 0.002 g / day to about 6 g / day. In some embodiments, the amount of the composition is about 0.005 g / day to about 5 g / day. In some embodiments, the amount of the composition is about 0.01 g / day to about 5 g / day. In some embodiments, the amount of the composition is about 0.02 g / day to about 5 g / day. In some embodiments, the amount of the composition is about 0.05 g / day to about 2.5 g / day. In some embodiments, the amount of the composition is about 0.1 g / day to about 1 g / day. In some embodiments, dose levels below the lower limit of the above range may be sufficient. In some embodiments, dose levels above the upper limit of the above range may be required. In some embodiments, the composition is administered as a single dose once daily. In some embodiments, the composition is administered as multiple doses more than once daily. In some embodiments, the composition is administered twice daily. In some embodiments, the composition is administered three times daily. In some embodiments, the composition is administered four times daily. In some embodiments, the composition is administered more than four times daily. In some embodiments, the pharmaceutical composition is administered to a mammal. In some embodiments, the mammal is a human.

[0052] Examples 1-6 and Comparative Examples 1-2 of this invention at least support the protection scope of "an Enterococcus faecalis composition comprising: Enterococcus faecalis and lactoferrin, wherein the Enterococcus faecalis was deposited at the Guangdong Provincial Center for Microbial Culture Collection on October 13, 2022, with accession number GDMCC NO. 62869".

[0053] Examples 1-6 of this invention respectively support the protection scope of "the weight ratio of lactoferrin to Enterococcus faecalis is 10-600:1", "the weight ratio of lactoferrin to Enterococcus faecalis is 15-400:1", and "the weight ratio of lactoferrin to Enterococcus faecalis is 15-250:1".

[0054] The present invention has at least the following beneficial effects: Compared with existing technologies, this invention, through the combination of specific bacterial strains and lactoferrin, achieves significant improvements in the following indicators at lower doses: (1) Decreased stool score The experimental animals showed a significant reduction in diarrhea symptoms, indicating a marked improvement in their intestinal condition and an improvement in different types of diarrhea.

[0055] (2) Optimization of gut microbiota abundance Specific strains of Enterococcus faecalis, in synergy with lactoferrin, enhance iron chelation efficiency, depriving harmful bacteria of essential nutrients for growth while creating conditions for beneficial bacteria to colonize. This helps improve gut microbiota imbalance, irritable bowel syndrome, and radiation proctitis, and aids in restoring the balance of the gut microbiota.

[0056] (3) Downregulation of TNF-α expression and recovery of sIgA levels It significantly reduced the expression level of the pro-inflammatory factor TNF-α in the intestine, alleviated the intestinal inflammatory response, and after 28 days of administration, the local sIgA in the intestine decreased significantly, indicating that the infection was controlled and the mucosal immune status returned to homeostasis.

[0057] Furthermore, based on the present invention: Based on the comparison of Examples 1-6 and Comparative Examples 1-2, it is shown that the weight ratio of specific Enterococcus faecalis strains to lactoferrin is the key to synergy. Under appropriate weight ratios and bacterial contents, the efficacy of regulating immunity and treating intestinal diseases can be significantly improved.

[0058] Based on the comparison of Examples 1-6 and Comparative Examples 3-5, it is shown that specific Enterococcus faecalis strains are key to synergistic effects with lactoferrin, which can significantly improve the efficacy of regulating immunity and treating intestinal diseases. Attached Figure Description

[0059] Figure 1 This is a diagram illustrating the fecal scoring criteria.

[0060] Figure 2 This is a diagram of the 16S sequencing results of the bacterial community. Detailed Implementation

[0061] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0062] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0063] In the following examples, Enterococcus faecalis ( Enterococcus faecalisThe strain is Enterococcus faecalis HHP003, disclosed in patent CN117511773A, and deposited at the Guangdong Provincial Center for Microbial Culture Collection on October 13, 2022, with accession number GDMCC NO. 62869.

[0064] The comparative strains Enterococcus faecalis ATCC51299, Enterococcus faecalis ATCC29212, and Enterococcus faecalis CICC20419 were all commercially available.

[0065] In the following examples, all Enterococcus faecalis used were prepared as Enterococcus faecalis bacterial suspensions: the bacterial suspensions were first cultured to the logarithmic phase, then subjected to heat inactivation treatment (60°C for 30 min), followed by ultraviolet irradiation inactivation for 15 min. After inactivation, an appropriate amount of bacterial suspension was spread on a culture medium plate for incubation to verify that there were no viable bacteria residues. After confirming complete inactivation, the inactivated bacterial suspensions were used for subsequent cell experiments and animal experiments. The viable bacterial counts (bacterial amounts) of the inactivated bacterial suspensions before inactivation were 1 billion CFU / g, 800 billion CFU / g, and 1 trillion CFU / g, respectively.

[0066] Lactoferrin was purchased from Saputo Dairy Australia Pty Ltd, product number 1001102, and is bovine lactoferrin.

[0067] Examples 1-6, Comparative Examples 1-2 Examples 1, 3-5, and Comparative Examples 1-2 used bacterial suspensions with a bacterial count of 800 billion CFU / g, Example 2 used a bacterial suspension with a bacterial count of 1 billion CFU / g, and Example 6 used a bacterial suspension with a bacterial count of 1 trillion CFU / g.

[0068] In Examples 1-6 and Comparative Examples 1-2, the *Enterococcus faecalis* used was deposited at the Guangdong Provincial Center for Microbial Culture Collection on October 13, 2022, with accession number GDMCC NO. 62869. The weights of lactoferrin and *Enterococcus faecalis* suspensions are shown in the table below.

[0069] Table 1

[0070] Comparative Example 3 Unlike Example 5, the Enterococcus faecalis strain used was the control strain Enterococcus faecalis ATCC51299.

[0071] Comparative Example 4 Unlike Example 5, the Enterococcus faecalis strain used was the control strain Enterococcus faecalis ATCC29212.

[0072] Comparative Example 5 Unlike Example 5, the Enterococcus faecalis strain used was the control strain Enterococcus faecalis CICC20419.

[0073] Result detection 1. Modeling and drug administration: This experiment selected healthy British Shorthair Blue cats of similar weight and approximately one year of age as test animals. All test animals were housed in a temperature-controlled environment, fed the same daily diet, and provided with free access to water throughout the experiment. Before the start of the experiment, all test animals underwent a 7-day pre-feeding program to alleviate stress caused by transportation and environmental changes.

[0074] Modeling method: Enterotoxigenic Escherichia coli (ETEC) was administered via gavage for 8 days to establish the model, with a dosage of 1 billion CFU / kg / day; Intervention method: After the modeling was completed, the animals were randomly divided into groups of 8 each. The intervention was started and continued for 28 days. The combination of the example and the comparative example was administered by gavage once a day, with a gavage volume of 5 mL.

[0075] Normal group: Normal animals were given PBS (phosphate-buffered saline) by gavage daily. Model group: Model animals, administered PBS by gavage daily; Lactoferrin group: model animals, administered 300 mg / animal / day by gavage; Enterococcus faecalis group: model animals, administered 30 mg / animal / day by gavage; Combined group: Model animals, administered 150 mg / animal / day of the combination of the example and the comparative example by gavage.

[0076] 2. Test Results (1) Stool scoring, the specific method is as follows: Fecal samples were collected, and fecal scores were performed every two days, with photographs taken to record the fecal condition. Fecal scores were based on a cat fecal characteristics rating scale of 1-9, primarily determined by characteristics such as fecal formation, consistency, moisture content, and ease of collection. This was used to evaluate the gastrointestinal function during the experiment and the effects of different treatments on fecal characteristics.

[0077] Stool scoring criteria reference Figure 1 .

[0078] The results for normal animals and model animals before and after modeling are as follows: Table 2

[0079] Note: Compared with before modeling, after modeling. # P<0.05 The results showed a significant increase in fecal scores, indicating successful model establishment.

[0080] The results after 28 days of intervention are as follows: Table 3

[0081] Note: Compared with the normal group, the model group is different. ## P<0.01; compared with the model group, P<0.01, P<0.05; compared with the comparative example group and Example 5 group, a P<0.05, aa P<0.01 (2) Blood indicators: inflammatory factors (TNF-α), immune cell markers (CD3+) + FOXP3 + The specific method is as follows: Collect 2 mL of blood sample and store it in a regular blood collection tube. Centrifuge at 5000 r / min for 10 min, separate the supernatant, and detect it using the ELISA method. The results for normal animals and model animals before and after modeling are as follows: Table 4

[0082] Note: Compared with before modeling, after modeling. ## P<0.01 After modeling, inflammatory factors (TNF-α) and immune cell markers (CD3+) were observed. + FOXP3 + () significantly increased.

[0083] The results after 28 days of intervention are as follows: Table 5

[0084] Note: Compared with the normal group, the model group is different. ## P<0.01; compared with the model group, P<0.01, P<0.05; compared with the comparative example group and Example 5 group, a P<0.05, aa P<0.01 (3) Fecal index: sIgA, the determination method is as follows: A 2g fecal sample was collected, rapidly frozen in a liquid nitrogen tank, and then detected using the ELISA method.

[0085] The normal group had an sIgA content of 67.03±41.94 μg / mL, which decreased to 64.49±39.50 μg / mL after modeling. The model animals had an sIgA content of 68.99±33.31 μg / mL before modeling, which decreased to 96.16±52.25 μg / mL after modeling. The significant increase in sIgA content indicates successful modeling.

[0086] The results after 28 days of intervention are as follows: Table 6

[0087] Note: Compared with the normal group, the model group is different. ## P<0.01; compared with the model group, P<0.01, P<0.05; compared with the comparative example group and Example 5 group, a P<0.05, aa P<0.01 (4) Taking Example 5 as an example, the bacterial community was further sequenced using 16S sequencing. The sequencing results are as follows: Figure 2 As shown, the results indicate that the abundance of some bacterial communities was significantly increased, suggesting that the combination in Example 5 has the effect of regulating the intestinal flora.

[0088] Results Analysis Experimental results showed that after intervention with a combination of specific Enterococcus faecalis and lactoferrin, the fecal scores of experimental animals decreased and diarrhea symptoms were significantly alleviated, indicating a significant improvement in intestinal condition and optimization of intestinal flora abundance. At the same time, the expression level of the pro-inflammatory factor TNF-α in the blood was reduced, and the inflammatory response was relieved. After 28 days of intervention, local sIgA in the intestine decreased significantly, indicating that the combination of the present invention has good effects in regulating immunity and improving intestinal health.

[0089] Meanwhile, compared with the single Enterococcus faecalis intervention group and the single lactoferrin intervention group, the combined intervention group showed significant advantages in terms of fecal score and reduction of inflammatory response; the results of comparative examples 3-5 indicate that the selection of specific Enterococcus faecalis is the key to its synergistic effect with lactoferrin.

[0090] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A composition of Enterococcus faecalis, characterized in that, include: Enterococcus faecalis and lactoferrin, wherein the Enterococcus faecalis was deposited at the Guangdong Provincial Center for Microbial Culture Collection on October 13, 2022, with accession number GDMCC NO. 62869, the weight ratio of lactoferrin to Enterococcus faecalis is 10-600:1, and the bacterial count of Enterococcus faecalis is 1 billion CFU / g-1 trillion CFU / g.

2. The Enterococcus faecalis composition according to claim 1, characterized in that, The weight ratio of lactoferrin to Enterococcus faecalis is 15-400:

1.

3. The Enterococcus faecalis composition according to claim 2, characterized in that, The weight ratio of lactoferrin to Enterococcus faecalis is 15-250:

1.

4. The Enterococcus faecalis composition according to any one of claims 1-3, characterized in that, The bacterial count of Enterococcus faecalis was 800 billion CFU / g.

5. The Enterococcus faecalis composition according to any one of claims 1-3, characterized in that, The Enterococcus faecalis is the bacterial cell of Enterococcus faecalis or its preparation, and the preparation is selected from at least one of fermentation broth, fermentation broth precipitate, fermentation broth supernatant, live bacteria, inactivated bacteria, lyophilized powder, lysate, lysate, secondary metabolites, and exosomes.

6. An active formulation, characterized in that, The composition of Enterococcus faecalis according to any one of claims 1-5.

7. The active formulation according to claim 6, characterized in that, The dosage form of the active preparation is selected from decoctions, ointments, pills, lozenges, tablets, granules, powders, capsules, patches, oral liquids, or syrups.

8. The use of the Enterococcus faecalis composition according to any one of claims 1-5 or the active formulation according to any one of claims 6-7 in the preparation of products for the prevention and / or treatment of intestinal diseases.

9. The application according to claim 8, characterized in that, The intestinal diseases mentioned include indigestion, malabsorption, vomiting, diarrhea, dysbiosis, irritable bowel syndrome, and inflammatory bowel disease.

10. The use of the Enterococcus faecalis composition according to any one of claims 1-5 or the active formulation according to any one of claims 6-7 in the preparation of products that modulate immunity.

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