Application of plant lactobacillus in preparation of preparation for preventing and / or treating allergic rhinitis

Abstract

The invention relates to application of plant lactobacillus in preparation of a preparation for preventing and / or treating allergic rhinitis. The plant lactobacillus comprises Lactobacillus plantarum BGI-N6, the Lactobacillus plantarum BGI-N6 is preserved in the China Center for Type Culture Collection on June 5, 2023, and the preservation number of the Lactobacillus plantarum BGI-N6 is CCTCC (China Center for Type Culture Collection) NO: M2023948. Based on an allergic rhinitis animal model, the action mechanism of the plant lactobacillus in the inflammation process is deeply analyzed, it is found that the plant lactobacillus can effectively prevent and / or treat the allergic rhinitis, and a new strategy is provided for developing a scheme for treating the allergic rhinitis.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology and relates to the application of Lactiplantibacillus plantarum in the preparation of a preparation for preventing and / or treating allergic rhinitis. Background Art

[0002] Allergic rhinitis is a chronic inflammatory disease of the nasal mucosa mediated by immunoglobulin E (IgE). Its pathological process is manifested as an abnormal immune response jointly activated by immune active cells and cytokines after a susceptible individual contacts an allergen. Uncontrolled allergic rhinitis for a long time not only causes repeated nasal symptoms in patients (the triad of sneezing, nasal congestion, and runny nose), but also easily induces enhanced Th2-type immune polarization, forming a vicious cycle of "immune imbalance - inflammation aggravation", thus causing multi-dimensional health hazards including limited social function, decreased work efficiency, and impaired cognitive development.

[0003] The existing treatment methods for allergic rhinitis mainly include drug ladder therapy, immunotherapy, and probiotic intervention therapy. The drug treatment system follows a stepped plan. Among them, antihistamines can quickly relieve symptoms such as nasal itching and sneezing by blocking histamine receptors; local or systemic glucocorticoids can effectively inhibit the inflammatory reaction of the nasal mucosa; leukotriene receptor antagonists help improve nasal ventilation and nocturnal symptoms; mast cell stabilizers can reduce the release of inflammatory mediators; short-term use of decongestants can effectively relieve nasal congestion. For patients with a long-term course of disease, allergen-specific immunotherapy (ASIT) induces immune tolerance through progressive sublingual or subcutaneous exposure, thereby achieving a lasting remission of symptoms. In emerging treatment strategies, probiotic therapy has become an alternative by regulating the balance of the gut microbiota-immune axis, secreting anti-inflammatory factors, and enhancing mucosal barrier function.

[0004] Although the existing therapies have significant efficacy for allergic rhinitis symptoms, there are also obvious clinical limitations. The risks of long-term drug use are relatively prominent. For example, the first-generation antihistamines are prone to cause drowsiness and cognitive impairment; local hormones may cause nasal mucosa atrophy and fungal infections; if decongestants are used for more than 5 days, they may induce rebound congestion. Although immunotherapy has the potential for radical cure, its 3-5-year treatment course and high annual cost limit its popularity. The emerging probiotic intervention therapy is restricted by strain specificity. Different strains of the same species have significant differences in efficacy, and the baseline differences in the gut microecology make it difficult to predict individual responses, and the dose-effect relationship has not been clarified.

[0005] In summary, there is an urgent need for a more effective and safer treatment plan to improve the quality of life of patients with allergic rhinitis and reduce side effects during the treatment process. Summary of the Invention

[0006] In view of the deficiencies of the prior art and the actual needs, the present invention provides the application of Lactobacillus plantarum in the preparation of a preparation for preventing and / or treating allergic rhinitis, deeply analyzes Lactobacillus plantarum BGI-N6, and develops its application in the field related to allergic rhinitis.

[0007] To achieve this purpose, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides the application of Lactobacillus plantarum in the preparation of a preparation for preventing and / or treating allergic rhinitis; the Lactobacillus plantarum includes Lactobacillus plantarum BGI-N6, which was deposited at the China Center for Type Culture Collection on June 5, 2023, and the deposit number is CCTCC NO: M2023948.

[0009] In the present invention, based on an allergic rhinitis animal model, the mechanism of action of Lactobacillus plantarum BGI-N6 in the inflammatory process was deeply analyzed, and it was found that it can effectively prevent and / or treat allergic rhinitis, providing a new strategy for developing a treatment plan for allergic rhinitis.

[0010] It can be understood that the preparation may be a related preparation containing the cells of the Lactobacillus plantarum, such as freeze-dried bacterial powder, etc.

[0011] In a second aspect, the present invention provides a probiotic preparation for preventing and / or treating allergic rhinitis, and the probiotic preparation includes the Lactobacillus plantarum described in the first aspect.

[0012] Preferably, the concentration of the Lactobacillus plantarum in the probiotic preparation is 10 7 ~10 9 CFU / mL.

[0013] Preferably, the probiotic preparation further includes a pharmaceutically acceptable excipient.

[0014] Preferably, the excipient includes at least one of a carrier, a protective agent, a filler, a wetting agent, a solubilizer, an osmotic pressure regulator, a pH regulator, an antioxidant, or a buffer.

[0015] In a third aspect, the present invention provides the application of the Lactobacillus plantarum described in the first aspect in the preparation of a preparation for repairing nasal mucosa damage.

[0016] In the present invention, it was found that Lactobacillus plantarum BGI-N6 can effectively repair nasal mucosa tissue damage and improve the adverse pathological state of nasal mucosa tissue.

[0017] In a fourth aspect, the present invention provides the application of the Lactobacillus plantarum described in the first aspect in the preparation of a preparation for inhibiting intestinal pathogenic bacteria.

[0018] Preferably, the intestinal pathogenic bacteria include at least one of Escherichia coli, Staphylococcus aureus, Enterobacter cloacae or Pseudomonas aeruginosa.

[0019] In the present invention, it is found that Lactiplantibacillus plantarum BGI-N6 can effectively inhibit intestinal pathogenic bacteria such as Escherichia coli and Staphylococcus aureus, and has potential probiotic effects of maintaining the immune health of the body by regulating the balance of intestinal flora. It can be used for the development of related therapeutic drugs, and can also be used for the development of in vitro bacteriostatic agents for non-therapeutic purposes.

[0020] In the fifth aspect, the present invention provides the use of the Lactiplantibacillus plantarum described in the first aspect in the preparation of a preparation for regulating the levels of serum cytokines and / or intestinal permeability markers.

[0021] Preferably, the cytokines include at least one of interleukin-13 (IL-13), interferon-γ (IFN-γ) or transforming growth factor-β (TGF-β).

[0022] Preferably, the intestinal permeability markers include lipopolysaccharide (LPS) and / or D-lactic acid.

[0023] In the present invention, it is found that Lactiplantibacillus plantarum BGI-N6 can effectively regulate the levels of serum cytokines, reduce the level of IL-13, and increase the levels of IFN-γ and TGF-β. It can be used for the development of related therapeutic drugs, and can also be used for the development of cytokine level regulators for non-therapeutic purposes for basic mechanism research.

[0024] In the present invention, it is found that Lactiplantibacillus plantarum BGI-N6 can effectively regulate the levels of intestinal permeability markers, reduce the levels of LPS and D-lactic acid, and D-lactic acid and the like are produced by the metabolism of intestinal bacteria. The increase in its serum level reflects the damage of intestinal epithelial tight junctions. Therefore, BGI-N6 can be used for the development of drugs related to improving intestinal barriers, and can also be used for the development of regulators for non-therapeutic purposes for basic mechanism research.

[0025] In the sixth aspect, the present invention provides the use of the Lactiplantibacillus plantarum described in the first aspect in the preparation of a preparation for regulating the levels of serum allergy markers.

[0026] Preferably, the serum allergy markers include at least one of immunoglobulin E (IgE), platelet-activating factor (PAF) or leukotriene C4 (LCT4).

[0027] In the present invention, it is found that Lactiplantibacillus plantarum BGI-N6 can effectively regulate the levels of serum allergy markers, reduce the levels of IgE, LCT4, and PAF, and can be used to develop related therapeutic drugs (such as regulating the Th1 / Th2 balance, inhibiting IgE-mediated type I hypersensitivity reaction, and blocking the pro-inflammatory pathways of PAF and LCT4), and can also be used to develop regulators for regulating the levels of serum allergy markers for non-therapeutic purposes for basic mechanism research and the like.

[0028] In a seventh aspect, the present invention provides the use of the Lactiplantibacillus plantarum described in the first aspect in the preparation of a preparation for regulating the differentiation of CD4 + T lymphocytes.

[0029] In the present invention, it is found that Lactiplantibacillus plantarum BGI-N6 can effectively regulate the differentiation of CD4 + T lymphocytes, reduce the ratio of the number of Th17 / Treg cells, and can be used to develop related therapeutic drugs, and can also be used to develop regulators for non-therapeutic purposes for basic mechanism research and the like.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] (1) The Lactiplantibacillus plantarum BGI-N6 provided by the present invention has no potential harm to the body, has good safety, and has no toxic and side effects compared with drug intervention, and avoids the side effects brought by drug intervention, and is suitable for long-term use;

[0032] (2) The Lactiplantibacillus plantarum BGI-N6 provided by the present invention has good ability to inhibit intestinal pathogenic bacteria and has the probiotic potential to maintain normal immune function by balancing the intestinal flora;

[0033] (3) The Lactiplantibacillus plantarum BGI-N6 provided by the present invention can repair the intestinal barrier, reduce the systemic exposure of allergens; reduce the levels of pro-inflammatory mediators platelet-activating factor (PAF), leukotriene C4 (LTC4), and IgE levels, regulate the balance of spleen T lymphocyte differentiation helper T cell 17-regulatory T cell (Th17-Treg), etc., effectively improve or relieve allergic rhinitis symptoms such as sneezing, nose rubbing, and runny nose, and have the probiotic effect of repairing the nasal mucosa damage caused by allergic rhinitis;

[0034] (4) The present invention explores the new functions of Lactiplantibacillus plantarum BGI-N6, provides a new direction for the prevention and / or treatment of allergic rhinitis, and can be applied to the fields of food, health products, and pharmaceuticals, and has broad industrial prospects. Description of the Drawings

[0035] Figure 1The symptom scores of allergic rhinitis in rats of each group in Example 6, including typical rhinitis symptoms such as sneezing, nose rubbing, and runny nose (* indicates p < 0.05 compared with the model control group; ** indicates p < 0.01 compared with the model control group; *** indicates p < 0.001 compared with the model control group; tested by ONE-WAY ANOVA).

[0036] Figure 2 The levels of allergy markers IgE, PAF, LCT4, and Th2-type cytokine IL-13 in the sera of rats of each group in Example 6 (* indicates p < 0.05 compared with the model control group; ** indicates p < 0.01 compared with the model control group; *** indicates p < 0.001 compared with the model control group; tested by ONE-WAY ANOVA).

[0037] Figure 3 The levels of Th1-type cytokine IFN-γ, Treg-type cytokine TGF-β, and intestinal barrier damage markers LPS and D-lactic acid in the sera of rats of each group in Example 6 (* indicates p < 0.05 compared with the model control group; ** indicates p < 0.01 compared with the model control group; *** indicates p < 0.001 compared with the model control group; tested by ONE-WAY ANOVA)

[0038] Figure 4 For the CD4 + T lymphocyte count results in the spleens of rats of each group in Example 6, including the ratio of the numbers of Th17 and Treg cells (* indicates p < 0.05 compared with the model control group; ** indicates p < 0.01 compared with the model control group; *** indicates p < 0.001 compared with the model control group; tested by ONE-WAY ANOVA).

[0039] Figure 5 The histopathological diagrams of hematoxylin-eosin (HE) staining of the nasal mucosa tissues of rats of each group in Example 6.

[0040] Figure 6 The histopathological diagrams of periodic acid-Schiff (PAS) staining of the nasal mucosa tissues of rats of each group in Example 6. Detailed implementation manners

[0041] The technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and through specific implementation manners. However, the following examples are merely simple examples of the present invention and do not represent or limit the scope of the protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

[0042] For specific technologies or conditions not specified in the examples, follow the technologies or conditions described in the literature in this field or the product instructions. For reagents or instruments whose manufacturers are not specified, they are all conventional products that can be obtained by purchasing through regular channels.

[0043] The information on the main experimental reagents and consumables involved in the following examples is as follows:

[0044] Lactiplantibacillus plantarum BGI-N6 was deposited at the China Center for Type Culture Collection (CCTCC) on June 5, 2023, with the deposit number CCTCC NO: M2023948; the address of the deposit unit is No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, and the postal code is 430072.

[0045] Example 1

[0046] This example prepares the active probiotic powder of Lactiplantibacillus plantarum BGI-N6.

[0047] Preparing the active probiotic powder of Lactiplantibacillus plantarum BGI-N6 is a key step for its application. The preparation of the active probiotic powder of Lactiplantibacillus plantarum BGI-N6 includes two parts: high-density fermentation and freeze-drying.

[0048] High-density fermentation: Take the cryopreservation tube of Lactiplantibacillus plantarum BGI-N6 strain, inoculate it into 300 mL of MRS liquid medium (product number 027312, purchased from Guangdong Huankai Microbial Technology Co., Ltd.) at an inoculation amount of 1%, anaerobically culture at 37 °C for 24 h, and then inoculate it into 5 L of MRS liquid medium at an inoculation amount of 5%, anaerobically culture at 37 °C. When the cell density in the fermentation broth reaches 3.0×10 9 CFU / mL, collect the fermentation broth for the subsequent steps.

[0049] Freeze-drying: Centrifuge the fermentation broth at 4000 rpm for 10 min at 4 °C, collect the bacterial sludge, add 60 g of skim milk powder, 120 g of trehalose, 20 g of maltodextrin, 8 g of glycerol, and 1 g of sodium ascorbate as a freeze-drying protectant, add 800 mL of sterile water to dissolve and mix evenly, pre-freeze at -60 °C for 3 h, and then place it in a freeze-dryer. Complete the freeze-drying according to the instructions of the freeze-dryer to finally obtain the active probiotic powder of Lactiplantibacillus plantarum BGI-N6, and its active probiotic content is 200 billion CFU / g.

[0050] Example 2

[0051] This example evaluates the inhibitory ability of Lactiplantibacillus plantarum BGI-N6 against intestinal pathogenic bacteria.

[0052] The gut microbiota is closely linked to the human immune system, and the balance of the gut microbiota is crucial for maintaining the normal function of the immune system. By measuring the inhibitory ability of Lactiplantibacillus plantarum BGI-N6 against common intestinal pathogens in vitro, it helps to understand its regulatory effect on the balance of the gut microbiota.

[0053] Brain Heart Infusion Broth Medium (BHI, product number HB8297-1, purchased from Qingdao Haibo Biotechnology Co., Ltd.) was used to activate 4 common intestinal pathogenic bacteria, including Escherichia coli ATCC25922, Staphylococcus aureus ATCC29213, Enterobacter cloacae ATCC13047, and Pseudomonas aeruginosa ATCC27853. After activation, sterile Phosphate Buffered Saline (PBS, product number HB8507-1, purchased from Qingdao Haibo Biotechnology Co., Ltd.) was used to adjust the cell concentration OD 600 = 0.7 for standby, denoted as each pathogenic bacteria solution. MRS Broth Medium (product number 027312, Guangdong Huankai Microbial Sci & Tech Co., Ltd.) was used to activate the Lactiplantibacillus plantarum BGI-N6 active probiotic powder in Example 1. The bacterial solution was centrifuged at 7000 rpm for 5 min to obtain the probiotic fermentation supernatant, which was filtered through a 0.22 μm filter membrane and reserved for use, denoted as the probiotic fermentation supernatant.

[0054] The inhibitory ability of Lactiplantibacillus plantarum BGI-N6 against common intestinal pathogens was determined by the co-culture inhibition method: In the experimental group, 50 μL of each adjusted-concentration pathogenic bacteria solution, 50 μL of the probiotic fermentation supernatant, and 50 μL of double-concentration BHI broth medium were added to the 96-well plate; in the control group, an equal volume of sterile normal saline was used to replace the probiotic fermentation supernatant. After sample addition, the 96-well plate was placed in an incubator at 37 °C for 24 h, and the growth of different pathogenic bacteria was evaluated by OD 600 absorbance. The experiment was repeated 3 times, and the results were expressed as mean ± standard deviation. The pathogen inhibition rate (%) = [1 - (OD 600 experimental group / OD 600 control group)] × 100%

[0055] The experimental results of the inhibitory ability of Lactiplantibacillus plantarum BGI-N6 against intestinal pathogens are shown in Table 1. The growth inhibition rate of Lactiplantibacillus plantarum BGI-N6 against Escherichia coli was 88.93 ± 4.40%; the growth inhibition rate against Staphylococcus aureus was 95.70 ± 2.90%; the growth inhibition rate against Enterobacter cloacae was 94.25 ± 2.20%; the growth inhibition rate against Pseudomonas aeruginosa was 85.70 ± 5.60%.

[0056] Table 1 Inhibitory ability of Lactiplantibacillus plantarum BGI-N6 against intestinal pathogens

[0057] Pathogen species Inhibition rate (%) Escherichia coli 88.93±4.40 Staphylococcus aureus 95.70±2.90 Enterobacter cloacae 94.25±2.20 Pseudomonas aeruginosa 85.70±5.60

[0058] The above results indicate that Lactiplantibacillus plantarum BGI-N6 has extremely high inhibitory ability against common intestinal pathogens and has potential probiotic effects in maintaining the immune health of the body by regulating the balance of the intestinal flora.

[0059] Example 3

[0060] In this example, a low-dose probiotic agent for improving allergies was prepared.

[0061] The preparation method of the low-dose Lactiplantibacillus plantarum BGI-N6 probiotic agent for improving allergic reactions includes the following steps: accurately weigh 0.01 g of the freeze-dried powder of Lactiplantibacillus plantarum BGI-N6 (containing 200 billion CFU / g of active probiotics), dissolve it in 199.99 mL of sterilized physiological saline, and stir the solution until it is completely mixed to obtain a probiotic agent for improving allergies with a cell concentration of 1.0×10 7 CFU / mL for improving allergic reactions.

[0062] Example 4

[0063] In this example, a medium-dose probiotic agent for improving allergies was prepared.

[0064] The preparation method of the medium-dose Lactiplantibacillus plantarum BGI-N6 probiotic agent for improving allergic reactions includes the following steps: accurately weigh 0.10 g of the freeze-dried powder of Lactiplantibacillus plantarum BGI-N6 (containing 200 billion CFU / g of active probiotics), dissolve it in 199.90 mL of sterilized physiological saline, and stir the solution until it is completely mixed to obtain a probiotic agent for improving allergies with a cell concentration of 1.0×10 8 CFU / mL for improving allergic reactions.

[0065] Example 5

[0066] In this example, a high-dose probiotic agent for improving allergies was prepared.

[0067] The preparation method of the high-dose Lactiplantibacillus plantarum BGI-N6 probiotic agent for improving allergic reactions includes the following steps: accurately weigh 1.00 g of the freeze-dried powder of Lactiplantibacillus plantarum BGI-N6 (containing 200 billion CFU / g of active probiotics), dissolve it in 199.00 mL of sterilized physiological saline, and stir the solution until it is completely mixed to obtain a probiotic agent for improving allergies with a cell concentration of 1.0×10 9 CFU / mL for improving allergic reactions.

[0068] Comparative Preparation Example

[0069] In this comparative preparation example, an agent for improving allergies was prepared.

[0070] The preparation method of the medicament that can be used to improve allergic reactions includes the following steps: accurately weigh 10.4 mg of ground loratadine (product number ZH10970410, purchased from Bayer Healthcare Co., Ltd.), dissolve it in 50 mL of sterile physiological saline, and continuously stir until the solution is uniformly mixed to finally obtain an anti-allergy medicament with a drug concentration of 0.208 mg / mL.

[0071] Example 6

[0072] By constructing a rat model of allergic rhinitis, the efficacy of the control anti-allergy medicament and each dose of the anti-allergy probiotic agent prepared in Examples 3 - 5 in allergic rhinitis was verified as follows:

[0073] Take 36 SPF-grade female SD rats with a body weight of 300 ± 20 g. They are raised in groups of 6 per cage, and the breeding temperature and humidity are 20 - 26°C and 40 - 70% respectively, with 12h:12h day-night intermittent lighting; the conditions in the breeding room are always kept stable to ensure the reliability of the experimental results. The experiment is divided into 6 groups, including a normal control group (A), a model control group (B), a positive control group (loratadine, C), and low-dose (D), medium-dose (E), and high-dose intervention groups of Lactiplantibacillus plantarum BGI-N6 (F), with 6 rats in each group. The group and dose design are shown in Table 2.

[0074] Table 2 List of group and dose design

[0075]

[0076] The experimental stage is divided into three stages: the sensitization period (days 0 - 15), the intervention period (days 22 - 50), and the challenge period (days 42 - 50) (the day of intraperitoneal injection of the sensitizing reagent is recorded as day 0). An ovalbumin (OVA)-induced rat model of allergic rhinitis is used, where the sensitizing reagent contains 1 mg / mL OVA and 21 mg / mL Al(OH)3, and the challenge reagent contains 30 mg / mL OVA.

[0077] The preparation method of the sensitizing reagent is: weigh 10 mg of OVA and 210 mg of aluminum hydroxide, dissolve them in 10 mL of sterile physiological saline, and make the sensitizing reagent after mixing evenly.

[0078] The preparation method of the challenge reagent: weigh 300 mg of OVA, dissolve it in 10 mL of sterile physiological saline, and make the challenge reagent after mixing evenly.

[0079] During the sensitization period, except for the normal control group which was injected with 1 mL of sterile saline, the remaining groups of rats were intraperitoneally injected with 1 mL of the sensitizing reagent on days 0, 3, 6, 9, 12, and 15; during the intervention period, each dose intervention group of BGI-N6 was gavaged with different corresponding doses of the anti-allergy probiotic agent, with a daily gavage dose of 1 mL for 28 consecutive days. The normal control group and the model control group were given an equal amount of saline instead of the anti-allergy probiotic agent. The positive control group was given loratadine solution instead of the anti-allergy probiotic agent; during the challenge period, except for the normal control group, the remaining groups of rats were challenged with 0.1 mL / rat / naris of the challenge reagent by nasal drip on days 42, 44, 46, 48, and 50. The normal control group was dripped with an equal amount of sterile saline at the same time. After the last experiment was completed (day 50), the symptoms of nose rubbing, sneezing, runny nose, etc. of each rat within 10 minutes were observed and recorded, and the scoring was carried out according to the "Animal Allergic Rhinitis Symptom Scoring Table (Table 3)".

[0080] Table 3 Animal Allergic Rhinitis Symptom Scoring Table

[0081] Allergic symptoms Normal (0 points) Mild (1 point) Moderate (2 points) Severe (3 points) Sneezing None ≤ 3 times 4 - 10 times ≥ 11 times Rubbing the nose None Gently scratching the nose and face Often scratching the nose and face Acute friction Running nose None Flowing in front of the nostrils Flowing past the front nostrils Flowing on the face

[0082] After the allergic rhinitis symptom scoring was completed (day 50), the rats were euthanized (day 51), and then serum samples, nasal mucosa tissue samples, and spleen samples were collected, and the allergic-related biological indexes were measured.

[0083] Method for collecting nasal mucosa samples and detection content: Referring to the method described in the academic paper "Improvement of the method for nasal lavage and nasal mucosa acquisition in mice" published by Zhang Junyan et al. in the Journal of Otorhinolaryngology-Skull Base Surgery of China, the nasal mucosa tissue was obtained, fixed with paraformaldehyde and prepared for paraffin sections, and then hematoxylin-eosin (HE) staining and periodic acid-Schiff (PAS) staining were carried out to evaluate the pathological changes of the nasal mucosa.

[0084] Blood sample collection method and detection content: After collecting the blood samples of rats, let them stand still at room temperature for 60 min to coagulate, centrifuge at 3000 rpm for 10 min at 4 °C, aspirate the upper serum, and measure the levels of allergy markers immunoglobulin E (IgE, batch number MM-0063R1, Jiangsu enzyme immunoassay), platelet-activating factor (PAF, batch number MM-92804501, Jiangsu enzyme immunoassay), leukotriene C4 (LCT4, batch number MM-92804901, Jiangsu enzyme immunoassay), and cytokines interleukin-13 (IL-13, batch number MM-0085R1, Jiangsu enzyme immunoassay), interferon-γ (IFN-γ, batch number 0198R1, Jiangsu enzyme immunoassay), transforming growth factor-β (TGF-β, batch number MM-0181R1, Jiangsu enzyme immunoassay), D-lactic acid (MM-92725801, Jiangsu enzyme immunoassay), and lipopolysaccharide (LPS, batch number MM-92600101, Jiangsu enzyme immunoassay).

[0085] Spleen sample collection method and detection content: Quickly take out the spleen. Refer to the method described in the academic paper "Taurohyodeoxycholic acid alleviates trinitrobenzenesulfonic acid induced ulcerative colitis via regulating Th1 / Th2 and Th17 / Treg cells balance" published by Lv et al. in "Life Sciences". Use flow cytometry to count the CD4 + T lymphocytes in the rat spleen and calculate their proportions.

[0086] Test data processing method: All data are expressed in the form of mean ± standard deviation. Use ONE-WAY ANOVA test analysis to perform statistical analysis on the above test data. When p < 0.05, it indicates that there are significant differences between groups.

[0087] Results of anti-allergic efficacy experiment:

[0088] (1) The probiotic agent of Lactiplantibacillus plantarum BGI-N6 effectively improved the symptoms of allergic rhinitis. The rhinitis symptom score is an important basis for evaluating the severity of rat rhinitis symptoms. This score consists of dimensions such as sneezing, nose rubbing, and runny nose. There were significant behavioral differences among the rats in each group during the sensitization period. By Figure 1It can be seen that the allergic rhinitis scores such as sneezing, nose rubbing and rhinorrhea scores in the model control group were significantly higher than those in the normal control group, indicating that OVA sensitization successfully induced the allergic rhinitis phenotype. The symptom scores of the positive control group and the probiotic intervention group (groups D-F) showed a significant decrease. Among them, the score of the medium-dose intervention group decreased most significantly, and the effect was better than that of the positive control. It is suggested that Lactiplantibacillus plantarum BGI-N6 at different doses can effectively relieve allergic rhinitis and improve allergic symptoms.

[0089] (2) The probiotic agent of Lactiplantibacillus plantarum BGI-N6 effectively regulates the levels of serum allergy markers and Th2-type cytokines. IgE is the core mediator of type I hypersensitivity. After binding to the receptor on the surface of mast cells, it triggers degranulation, releases sensitizing factors such as histamine, and directly causes symptoms such as nasal itching and sneezing. IL-13 is secreted by Th2 cells and induces goblet cell hyperplasia, mucin overexpression and eosinophil infiltration through the STAT6 signaling pathway, and is the core driving factor for nasal congestion and rhinorrhea. Platelet-activating factor (PAF) and leukotriene C4 (LCT4) are metabolites of arachidonic acid, which synergistically induce vascular leakage, mucosal edema and eosinophil chemotaxis. It can be Figure 2 seen that compared with the normal control values, the levels of serum IgE, PAF, LCT4 and IL-13 in the model control group were significantly increased, reflecting the hyperactive Th2 immune response in allergic rhinitis rats. After intervention with the BGI-N6 probiotic agent for improving allergy at different doses described in Examples 3-5, the levels of serum IgE, PAF, LCT4 and IL-13 in rats showed a significant decrease, indicating that BGI-N6 can regulate the Th1 / Th2 balance, inhibit the IgE-mediated type I hypersensitivity reaction, and block the pro-inflammatory pathways of PAF and LCT4, so as to achieve the purpose of improving the symptoms of allergic rhinitis.

[0090] (3) The probiotic agent of Lactiplantibacillus plantarum BGI-N6 effectively regulates cytokines and improves the intestinal barrier. Allergic rhinitis is characterized by the over-activation of Th2 immune response. IFN-γ secreted by Th1 cells can antagonize Th2 response and inhibit the release of pro-inflammatory factors such as IL-13. Treg cells inhibit Th2 / Th17 inflammatory reactions by secreting cytokines such as TGF-β and maintain immune tolerance. D-lactic acid is produced by the metabolism of intestinal bacteria, and the increase in its serum level reflects the disruption of intestinal epithelial tight junctions. The leaked LPS in the intestine activates macrophages through pathways such as TLR4 / NF-κB, further promoting the release of pro-inflammatory factors and exacerbating Th2 immune deviation. It can be Figure 3It can be seen that in the allergic rhinitis rat model group, the Th1 marker IFN-γ and the Treg marker TGF-β were significantly decreased, while the intestinal permeability markers LPS and D-lactic acid were significantly increased, suggesting insufficient Th1 and Treg immune responses in allergic rhinitis rats and impaired intestinal barrier function. After intervention with the improved allergy BGI-N6 probiotic agent at different doses described in Examples 3-5, the levels of IFN-γ and TGF-β were significantly increased, and at the same time, LPS and D-lactic acid also showed a significant decreasing trend. This result indicates that BGI-N6 can effectively repair the intestinal barrier function, promote the differentiation of immunoregulatory T cells, and reverse the Th2-dominant immune deviation.

[0091] (4) The BGI-N6 probiotic agent of Lactiplantibacillus plantarum effectively regulated the differentiation of CD4 + T lymphocytes in the spleen of allergic rhinitis rats. Treg cells inhibit Th2 / Th17 differentiation by secreting cytokines such as TGF-β, and Th17 cells exacerbate eosinophil infiltration and mucus hypersecretion by secreting cytokines such as IL-17. It can be Figure 4 seen that the Th17 / Treg ratio in the model control group was significantly increased, indicating an imbalance in immune homeostasis. After intervention with the improved allergy BGI-N6 probiotic agent at different doses described in Examples 3-5, the Th17 / Treg ratio was significantly lower than that in the model group. This shows that BGI-N6 can improve immune balance by regulating the differentiation of CD4 + T lymphocytes in the spleen of rats.

[0092] (5) The BGI-N6 probiotic agent of Lactiplantibacillus plantarum effectively improved the pathological state of the nasal mucosa tissue in allergic rhinitis rats. Pathological analysis can effectively evaluate the improvement of the nasal mucosa injury in allergic rhinitis rats by probiotics. It can be seen from Figure 5 and Figure 6 that the nasal mucosa structure of the rats in the normal control group was clear, the tissue was intact, arranged orderly, the epithelial cells were intact, no obvious inflammatory cell infiltration was seen, and no glandular hyperplasia was observed; while in the model control group, the rats showed various adverse pathological changes such as loose arrangement and shedding of nasal mucosa epithelial cells, glandular hyperplasia, goblet cell hyperplasia, and epithelial cilia breakage; after intervention with the improved allergy BGI-N6 probiotic agent at different doses described in Examples 3-5, the arrangement of nasal mucosa epithelial cells in allergic rhinitis rats was significantly improved, the phenomena of glandular and goblet cell hyperplasia were significantly reduced, and the nasal mucosa injury was significantly alleviated.

[0093] In summary, the present invention deeply analyzed Lactiplantibacillus plantarum BGI-N6 and found that it has good intestinal pathogen inhibitory ability, can restore and maintain the normal function of the immune system by balancing the intestinal flora, can regulate the levels of serum allergy markers and cytokines, regulate cytokines and improve the intestinal barrier, and regulate CD4 +T lymphocyte differentiation alleviates nasal mucosa damage caused by allergies, effectively improves allergic rhinitis symptoms, and provides a new direction for the prevention / treatment of allergic rhinitis.

[0094] The applicant declares that the above description is only a specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily conceived by any person skilled in the art within the technical scope disclosed by the present invention fall within the protection scope and the disclosure scope of the present invention.

Claims

1. Use of Lactobacillus plantarum in the preparation of a preparation for preventing and / or treating allergic rhinitis; The Lactobacillus plantarum includes Lactobacillus plantarum BGI-N6, which was deposited at the China Center for Type Culture Collection on June 5, 2023, with the deposit number CCTCC NO: M2023948.

2. A microecological preparation for preventing and / or treating allergic rhinitis, characterized in that, The probiotic preparation includes the Lactobacillus plantarum described in claim 1; Preferably, the concentration of Lactiplantibacillus plantarum in the microecological preparation is 10 7 ~10 9 CFU / mL; Preferably, the probiotic preparation further includes a pharmaceutically acceptable excipient; Preferably, the excipient includes at least one of a carrier, a protective agent, a filler, a wetting agent, a solubilizer, an osmotic pressure regulator, a pH regulator, an antioxidant, or a buffer.

3. Use of the Lactobacillus plantarum described in claim 1 in the preparation of a preparation for repairing nasal mucosa damage.

4. Use of the Lactobacillus plantarum described in claim 1 in the preparation of a preparation for inhibiting intestinal pathogenic bacteria.

5. The application according to claim 4, wherein The intestinal pathogenic bacteria include at least one of Escherichia coli, Staphylococcus aureus, Enterobacter cloacae, or Pseudomonas aeruginosa.

6. Use of the Lactobacillus plantarum described in claim 1 in the preparation of a preparation for regulating the levels of serum cytokines and / or intestinal permeability markers.

7. The application according to claim 6, characterized in that, The cytokines include at least one of interleukin-13, interferon-γ, or transforming growth factor-β; Preferably, the intestinal permeability markers include lipopolysaccharide and / or D-lactic acid.

8. Use of the Lactobacillus plantarum described in claim 1 in the preparation of a preparation for regulating the levels of serum allergy markers.

9. The application according to claim 8, characterized in that The serum allergy markers include at least one of immunoglobulin E, platelet-activating factor, or leukotriene C4.

10. Use of the Lactiplantibacillus plantarum described in claim 1 in the preparation of a preparation for regulating the differentiation of CD4 + T lymphocytes.

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