Application of AKK bacteria in improvement of immunity and preparation of products for prevention, treatment and / or adjuvant treatment of rhinitis

Abstract

The invention discloses application of AKK bacteria in improving immunity and preparing products for preventing, treating and / or assisting in treating rhinitis, and belongs to the technical field of microorganisms. In order to solve the technical problems that in the prior art, treatment means for rhinitis cannot relieve symptoms for a long time, the single anti-inflammatory effect is limited, side effects are likely to be caused and the like, the invention provides a new Ackermann mucophila strain AKK IMMU, the AKK IMMU is preserved in Guangdong Microbial Culture Collection Center on November 3, 2025, the preservation number is GDMCC No: 67192, and the AKK IMMU is preserved in the Guangdong Microbial Culture Collection Center on November 2025. The strain can remarkably improve rhinitis model mouse nasal mucosa pathological damage, regulate serum oxidative stress indexes and regulate lung tissue inflammatory factors.

Description

TECHNICAL FIELD

[0001] The application belongs to the field of microbial technology, and particularly relates to application of AKK bacteria in preparation of a product for preventing and treating or assisting in treating rhinitis. BACKGROUND

[0002] Rhinitis is a very common clinical condition, mainly divided into allergic rhinitis and non-allergic rhinitis. The etiology of allergic rhinitis (AR) is closely related to the interaction of genetics, environment and immune system. It not only causes local symptoms such as nasal congestion, runny nose, sneezing and nasal itching, but also can cause sleep disorders, attention deficit and other problems, which seriously interferes with the daily life, work and study of patients. More seriously, allergic rhinitis is closely related to other allergic diseases such as asthma, significantly increasing the risk of developing asthma in patients, and bringing double threats to the physical and mental health of patients. In recent years, studies have shown that mouse models have important value in the study of the mechanism of rhinitis. They not only reflect the pathological characteristics of human rhinitis, but also can be used to evaluate the effect of new treatment methods. By analyzing the immune response, pathological changes and inflammatory factor levels of mice, the occurrence and development process of rhinitis can be better understood.

[0003] At present, the treatment of allergic rhinitis mainly depends on drugs such as antihistamines, glucocorticoids, etc. However, these drugs can only relieve symptoms and cannot fundamentally cure the disease, and long-term use may cause serious side effects such as hormone dependence, drug resistance, etc., which brings potential risks to the health of patients. In addition, although desensitization therapy is a potential cure, it has a long course, high cost, and is not effective for all patients, limiting its widespread application. Therefore, developing safe and effective new treatment methods or intervention means has become an urgent need in the field of prevention and treatment of allergic rhinitis.

[0004] In recent years, probiotics as a new type of biological intervention means have shown great potential in the prevention and treatment of allergic diseases. Probiotics are a class of active microorganisms that are beneficial to the host, which can regulate the balance of intestinal microecology, enhance the intestinal barrier function, regulate the immune system and reduce inflammation. Many studies have shown that specific probiotic strains can regulate the activity of immune cells, inhibit the production of allergic-related cytokines, and thus alleviate the symptoms of allergic diseases.

[0005] The relevant patent documents searched are as follows: CN114514028A, published on May 17, 2022, discloses a pharmaceutical composition containing mucinophilic Akkermansia bacteria EBAMDK19 strain or its culture or dry matter for effectively preventing or treating atopic diseases, which can produce the same level of prevention or treatment effect of atopic diseases as steroid drugs, and the atopic diseases are asthma, atopic dermatitis, urticaria, allergic rhinitis, anaphylaxis or food allergy.

[0006] The prior art represented by the foregoing documents at least has the following unsolved technical problems or defects: The prior art only has a general scheme for treating pan-atopic diseases with mucinophilic Akkermansia bacteria, and allergic rhinitis is only listed as an "enumerated item" of atopic diseases. No technical solutions are designed and experimentally verified for the specific pathological mechanism, core symptoms, local drug delivery requirements, and exclusive detection indicators of rhinitis. SUMMARY

[0007] The purpose of the present application is to provide: The application of AKK bacteria in improving immunity, preparing products for preventing and treating or adjuvant treating rhinitis, and related technologies, to solve the technical problems or combinations thereof in the prior art, such as non-persistent symptom relief, limited single anti-inflammatory effect, and easy side effects of the treatment means for rhinitis.

[0008] Terminology: Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter of the claims belongs. Unless otherwise indicated, all patents, patent applications, publications, and other documents cited herein are hereby incorporated by reference in their entirety. If there is a plurality of definitions for a term herein, the definition in this section prevails.

[0009] It should be understood that the above brief summary and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter of the present application. In the present application, the use of singular includes plural unless specifically stated otherwise. It should also be noted that the use of "or" or "and" means "and / or" unless otherwise stated. In addition, the use of the term "include" and other forms, such as "includes" and "including", are not limiting.

[0010] The definitions of standard chemical terms can be found in the reference "Microbiology, Higher Education Press, Shen Ping, Chen Xiangdong, ed.".

[0011] Unless otherwise specified, conventional methods within the scope of the art are used, such as ELISA kit for detecting inflammatory factors and neurotransmitters, RT-qPCR for detecting gene relative expression, and histopathology HE staining observation.

[0012] Unless specific definitions are provided, the use of each of the terms given herein follows the standard techniques of the respective field. For example, the use of a reagent kit can be performed according to the manufacturer's instructions, or according to a method known in the art or described herein. The techniques and procedures described or referenced herein can be performed by routine methods known in the art, substantially as described in various general and more specific references which are cited and discussed throughout this specification.

[0013] The term "optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, that the description includes instances where the event or circumstance occurs and instances where it does not. For example, "optional excipient" means "no excipient" or "excipient".

[0014] The term "heat-inactivated AKK IMMU" as used herein refers to the AKK IMMU strain obtained by recovering and culturing (37℃, 24h) in MRS liquid medium, expanding (37℃, 18h, 3% inoculation amount), centrifuging (4℃, 4000rpm, 10min) to collect the strain, and then heat-inactivating (80℃, 30min) the strain after thawing the AKK IMMU strain from a -80℃ cryovial.

[0015] The term "rhinitis animal model" as used herein refers to a C57BL / 6 mouse rhinitis model constructed by intraperitoneal injection of ovalbumin (OVA) for basic sensitization (0.1mg / mL OVA + 2mg / mL aluminum hydroxide mixed solution, 0.2mL / time, once every other day, a total of 7 times, 1-13 days) and OVA for nasal challenge (5% OVA in saline solution, 10μL / naris, once a day, a total of 7 times, 15-21 days), and the model mouse has a total nasal symptom score of more than 5.

[0016] The term "oxidative stress indicator" as used herein refers to an indicator that can reflect the oxidative damage and antioxidant capacity of the body, including malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), nitric oxide (NO).

[0017] The term "inflammatory factor" as used herein refers to a cytokine involved in inflammatory response, including interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-17A (IL-17A), interferon-γ (IFN-γ).

[0018] The term "immunoglobulin" as used herein refers to an antibody involved in immune response, including immunoglobulin G (IgG), immunoglobulin M (IgM).

[0019] In a first aspect, the present application provides a mucinophilic Akkermansia bacteria AKK IMMU, which was deposited with the Guangdong Microbial Culture Collection Center on November 3, 2025, and has a deposit number of GDMCC No: 67192. In a second aspect, the present application provides an application of the above-mentioned mucinophilic Akkermansia bacteria AKK IMMU in the preparation of a product for improving immunity.

[0020] In a third aspect, the present application provides an application of the above-mentioned mucinophilic Akkermansia bacteria AKK IMMU in the preparation of a product for treating and / or assisting in treating rhinitis.

[0021] Among them, the treatment and / or auxiliary treatment of rhinitis is achieved by at least one of the following mechanisms: (1) Inhibition of the synthesis and release of inflammatory factors, reduction of inflammatory damage to the mucosa, and repair of the epithelial structure; (2) Improvement of the function of the antioxidant system, reduction of the stimulation of oxidative damage to the nasal mucosa; (3) Inhibition of specific immune response, reduction of antibody production, and reduction of allergic reaction.

[0022] In a fourth aspect, the present application provides a product containing the above-mentioned mucinophilic Akkermansia bacteria AKK IMMU.

[0023] Among them, the product includes at least one of the culture solution, whole bacteria or fermentation broth of the mucinophilic Akkermansia bacteria AKK IMMU.

[0024] Among them, the product includes but is not limited to: food and medicine.

[0025] Among them, the food includes but is not limited to: health products, drinks, dairy products, solid snacks, fermented foods.

[0026] Among them, the health products include but are not limited to: probiotic capsules, probiotic powders, probiotic tablets.

[0027] Among them, the drinks include but are not limited to: fermented milk, lactic acid bacteria beverages, probiotic oral liquids.

[0028] Among them, the dairy products include but are not limited to: fermented milk, probiotic cheese, probiotic milk powder; the solid snacks include probiotic biscuits, probiotic cereals, probiotic pastries.

[0029] Among them, the fermented foods include but are not limited to: probiotic fermented bean products, probiotic fermented grain products Among them, the medicines include but are not limited to: probiotic preparations, compound preparations containing probiotics.

[0030] Among them, the application amount of the product is not less than 107 CFU / kg.

[0031] The product is preferably administered in an amount of 1 x 10 7 - 1 x 10 9 CFU / kg.

[0032] The dosage form of the product includes but is not limited to: a liquid dosage form, a gaseous dosage form, a solid dosage form or a semi-solid dosage form.

[0033] The liquid dosage form includes: a solvent type, an aromatic water agent, a tincture, an elixir, a colloidal solution, a gum solution, a suspension or a emulsion.

[0034] The gaseous dosage form includes: an aerosol or a spray.

[0035] The solid dosage form includes: a powder, a pill, a tablet or a film.

[0036] The semi-solid dosage form includes: an ointment, a suppository or a paste.

[0037] The dosage form is preferably at least one of: a liquid solution, a lyophilized powder, a tablet, a capsule, a granule, a spray, an oral dispersible tablet / film, a sublingual tablet / lozenge.

[0038] The product further comprises an excipient.

[0039] The excipient includes but is not limited to: any one or more of a binder, a filler, a disintegrant, a lubricant, a preservative, an antioxidant, a flavoring agent, an aromatic agent, a co-solvent, an emulsifying agent, a solubilizing agent or an osmotic pressure regulator.

[0040] The excipient is preferably at least one of: lactose, mannitol, methyl cellulose, benzoic acid, sorbic acid, food color and / or talc.

[0041] Based on further solving or simultaneously solving multiple technical problems of the technical problem of the present application, in the technical scheme provided in the third aspect of the present application, the preferred scheme includes: The application of Akkermansia muciniphila AKK IMMU in preparing a product for relieving low-calcium-diet-induced osteoporosis, wherein the dosage of Akkermansia muciniphila AKK IMMU in the product is 1 x 10 7 CFU / kg-1 x 10 9 CFU / kg, and the dosage form is a bacterial liquid agent. This technical scheme further solves the technical problems of "convenience of product use and accuracy of dosage" on the basis of solving the technical problem of "relieving low-calcium-diet-induced osteoporosis".

[0042] The present application has at least the following beneficial effects: 1. Compared with the prior art (such as loratadine treatment), the present application has better technical effects in improving the pathological damage of the nasal mucosa of the rhinitis model mice, regulating the serum oxidative stress indicators, and regulating the inflammatory factors of the lung tissue.

[0043] According to experimental tests, the present application (heat-inactivated AKK IMMU group) reduces the nasal symptom score of the rhinitis model mice from 6.67±0.82 of the model group to below 4.33±0.52; reduces the serum MDA content from the elevated level of the model group to close to the normal group (the normal group 0.83±0.41 corresponding to the MDA baseline level); reduces the lung tissue IL-1β content from the elevated level of the model group to close to the normal group (the normal group 0.83±0.41 corresponding to the IL-1β baseline level); and reduces the nasal mucosa epithelial hyperplasia and edema degree from “significant hyperplasia and severe edema” of the model group to “mild hyperplasia and mild edema” or below.

[0044] Based on the consideration of entering other countries, the present application also provides the following technical solutions: The present application provides a method for preventing and / or adjuvant treating rhinitis, comprising administering a therapeutically effective amount of the mucinophilic Akkermansia AKK IMMU to a subject.

[0045] The term “subject” includes a living organism (e.g., a mammal) that can elicit an immune response. Examples of subjects include humans, primates, cows, horses, goats, sheep, dogs, cats, mice, rats, rabbits, guinea pigs, pigs, and transgenic species thereof.

[0046] The term “treat” or “treatment” or “ameliorate” refers to the medical management of a disease, disorder, or unwanted condition in a subject. Therapeutic or prophylactic benefit includes improved clinical outcome; reduction or alleviation of symptoms associated with the disease, disorder, or unwanted condition; decrease in symptom frequency; improved quality of life; longer disease-free status; reduction in disease, disorder, or unwanted condition severity; disease state stabilization; delay of disease progression; remission; survival; prolonged survival; or any combination thereof.

[0047] The term “therapeutically effective amount” refers to a dosage that is pharmaceutically effective, i.e., an amount of active drug sufficient to significantly improve the condition without producing serious side effects. The dosage depends on many factors, such as the nature and severity of the disease to be prevented or treated, the gender, age, weight, character, and individual response of the patient or animal, the administration route, the administration frequency, the treatment purpose, and thus the dosage of the present application can vary widely.

[0048] DEPOSIT DESCRIPTION Deposited Strain:Akkermansia muciniphila AKK IMMU; Classification: Akkermansia muciniphila ; Accession No:67192; Preservation Time:2025-11-03; Preservation Unit:Guangdong Microbial Culture Collection Center; Preservation Unit Abbreviation:GDMCC; Preservation Address:5th Floor, Building 59, 100, Martyrs' Road, Guangzhou. BRIEF DESCRIPTION OF DRAWINGS

[0049] Figure 1 For the pathological changes of mouse nasal tissue.

[0050] Figure 2 For the serum oxidative damage indicators MDA, NO, GSH, CAT, SOD levels of mice, the values shown as a-d are the mean ± standard deviation, according to Ducan test, the average values of different letters in the same figure are significantly different (P<0.05) p<0.05 ).

[0051] Figure 3 For the inflammatory indicators of mouse lung tissue IL-1 β, IL-6, IL-10, IL-17A and IFN-γ , the values shown as a-d are the mean ± standard deviation, according to Ducan test, the average values of different letters in the same figure are significantly different (P<0.05) p< 0.05 ).

[0052] Figure 4 For the immune indicators IgG and IgM levels of mouse spleen tissue, the values shown as a-c are the mean ± standard deviation, according to Ducan test, the average values of different letters in the same figure are significantly different (P<0.05) p<0.05 ).

[0053] Figure 5 For the IL-1β, IL-4, IL-6, IL-10, IL-17A and IFN-γ and IFN-γ gene expression in mouse brain tissue, the values shown as a-d are the mean ± standard deviation, according to Ducan test, the average values of different letters in the same figure are significantly different (P<0.05) p<0.05 ). DETAILED DESCRIPTION

[0054] Unless otherwise specified, the various raw materials and reagents in the present application are purchased from commercial suppliers, and experiments are carried out according to the operating instructions. Unless otherwise specified, the instruments, equipment, devices, etc. used in the present application are conventional instruments, equipment, devices, etc. and experiments are carried out according to the operating instructions and the matching reagents.

[0055] In order to make the objects, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with examples. If the specific conditions are not specified in the examples, the conventional conditions or the conditions suggested by the manufacturers are adopted. If the manufacturers of all reagents or instruments are not specified, the conventional products available in the market are adopted. In order to better illustrate the present application, numerous specific details are given in the following specific embodiments. The specific embodiments described herein are only used to explain the present application, and do not constitute any limitation on the present application.

[0056] All experimental data were statistically analyzed by data statistical software, combined with single factor variance analysis and Duncan method multiple comparison; p＜0.05 was considered statistically significant. The data were expressed as mean ± standard deviation (SD), repeated 3 times.

[0057] Table 1 Reagents and purchase sources

[0058] Basic Example 1 Preparation of bacterial solution The strain AKK IMMU was taken out from the experimental strain cryopreservation tube at-80 degrees Celsius, and after thawing, 200 μL was inoculated into MRS liquid medium (8 mL) and incubated at 37 degrees Celsius for 24 hours. Then 3% of the inoculum was transferred to MRS liquid medium for further incubation for 18 hours. Centrifugation (4 degrees Celsius, 4000 rpm, 10 minutes) was performed to collect the bottom AKK IMMU strain, and physiological saline was used for resuspension to adjust the cell concentration to 1.0×10 9 CFU / kg·bw and 1.0×10 7 CFU / kg·bw order of magnitude.

[0059] Basic Example 2 Preparation of reagents Preparation of intraperitoneal injection solution: 1 mg of OVA powder and 20 mg of aluminum hydroxide powder were weighed and added to 10 ml of physiological saline under sterile environment in a clean bench, and shaken well. A mixed suspension with a final concentration of 0.1 mg / mL OVA + 2 mg / mL aluminum hydroxide was prepared, which was used for intraperitoneal injection in the basic sensitization stage.

[0060] Preparation of nasal challenge solution: 50 mg of OVA dry powder was weighed and placed in a 3 mL sterile vial, and 1 mL of sterile physiological saline was added and shaken well to prepare a 5% OVA physiological saline solution, which was used for nasal challenge.

[0061] Example 1 Construction of animal model 30 C57BL / 6 mice, male, SPF level, 6-8 weeks old, purchased from Hunan Slike Jingda Experimental Animal Co., Ltd. Animal production license number: SYXK(Xiang)2024-0019. Mice were raised in standard mouse cages in the laboratory animal room, with room temperature of 23±2℃ and relative humidity of 55±5%. Light was turned on once every 12 hours, and the mice were free to eat and drink. After one week of adaptive feeding, the mice were randomly divided into 5 groups, 6 in each group, and adaptive feeding was performed before drug administration. This study was approved by the Ethics Committee of the Collaborative Innovation Center for Children's Nutrition and Health Development of Chongqing Second Normal University (2024112502B).

[0062] (1) Normal group: no intervention.

[0063] (2) Model group: OVA intraperitoneal injection for basic sensitization and OVA nasal instillation for nasal challenge were used to establish AR animal model. The specific operation method was as follows: 1-13 days, intraperitoneal injection of mixed solution containing 0.1 mg / mL OVA+2 mg / mL aluminum hydroxide 0.2 mL, a total of seven times, for basic sensitization of mice, from the 15th day, 10 μL of 5% OVA solution was continuously instilled into the nostrils, a total of seven times, for nasal challenge, until the 21st day.

[0064] (3) Positive control group: OVA intraperitoneal injection for basic sensitization and OVA nasal instillation for nasal challenge were used to establish AR animal model, and loratadine (2 mg / kg) was given by gavage for 3 weeks during modeling. The specific operation method was as follows: 1-13 days, intraperitoneal injection of mixed solution containing 0.1 mg / mL OVA+2 mg / mL aluminum hydroxide 0.2 mL, a total of seven times, for basic sensitization of mice, from the 15th day, 10 μL of 5% OVA solution was continuously instilled into the nostrils, a total of seven times, for nasal challenge, until the 21st day. Loratadine (2 mg / kg) was given by gavage every other day from the 1st day to the 21st day.

[0065] (4) Heat-inactivated AKK IMMU low-dose group (1.0×10 7 CFU / kg·bw): OVA intraperitoneal injection for basic sensitization and OVA nasal instillation for nasal challenge were used to establish AR animal model, and probiotic AKK IMMU was given by gavage for 3 weeks during modeling. The specific operation method was as follows: 1-13 days, intraperitoneal injection of mixed solution containing 0.1 mg / mL OVA+2 mg / mL aluminum hydroxide 0.2 mL, a total of seven times, for basic sensitization of mice, from the 15th day, 10 μL of 5% OVA solution was continuously instilled into the nostrils, a total of seven times, for nasal challenge, until the 21st day. Heat-inactivated (80℃, 30min) 1.0×10 7AKK IMMU gavage of the order of 1.0 x 10

[0066] (5) Heat-inactivated AKK IMMU high-dose group (1.0 x 10 9 AR animal model was established by intraperitoneal injection of OVA for basic sensitization and OVA nose drop for nasal challenge, and the probiotic AKK IMMU was given by gavage for 3 weeks during the modeling period. The specific operation method was as follows: 0.2 ml of mixed solution containing 0.1 mg / mL OVA + 2 mg / mL aluminum hydroxide was injected intraperitoneally every other day from 1-13 days, a total of seven times, to perform basic sensitization of mice. From the 15th day, 5% OVA solution of 10 μL was continuously dropped into the nostrils for 7 times per day to perform nasal challenge until the 21st day. At the same time, from the 1st day to the 21st day, heat-inactivated (80°C, 30 min) 1.0 x 10 9 AKK IMMU gavage of the order of 1.0 x 10

[0067] After the last nasal challenge, each mouse was placed in an individual cage for 30 minutes of observation, and the occurrence and severity of symptoms such as scratching the nose, sneezing, and running nose of the mouse were observed. The mouse was scored according to the nasal symptoms, and the scoring criteria were as follows: (1) Itchy nose: 1 point for a few times of light rubbing the nose, 2 points for repeated scratching of the nose, and 3 points for rubbing the nose all over the face. (2) Sneezing: 1-3 points for 1 point, 4-10 points for 2 points, and more than 11 points for 3 points. (3) Clear nasal discharge: 1 point for running to the anterior naris, 2 points for more than the anterior naris, and 3 points for running to the face. The scores of each nasal symptom were recorded by using the additive quantification method, and then the total score of each mouse was calculated. If the total score was more than 5, the AR model was successfully established and could be used for subsequent experiments.

[0068] After weighing, the mice were anesthetized with ether, and blood was taken after enucleation. Then the mice were sacrificed by the cervical dislocation method. After removing the nasal hair, the nose was cut off, and after decalcification, it was fixed in the tissue fixing fluid for more than 24 hours. The lung and spleen tissues were dissected and stored at -80°C for subsequent experiments.

[0069] The results of clinical symptom observation and body weight change of mice are shown in Table 2.

[0070] Table 2 Clinical symptom observation and body weight change

[0071] a-d The values shown are the mean ± standard deviation. According to the Ducan test, the mean values of different letters in the same column are significantly different (P<0.05). p<0.05 .

[0072] .Compared with the normal group, the symptom scores of all model groups were ≥5 points, indicating that the mouse AR model was successfully established in this study, and the differences were statistically significant. p<0.05 Compared with the model group, the symptom scores of the positive control group and the heat-inactivated L-AKK IMMU group and H-AKK IMMU group were reduced ( p<0.05 ), of which the positive control group was more effective, and the symptom score of the H-AKK IMMU group had no statistically significant difference with that of the L-AKK IMMU group ( p>0.05 ).

[0073] As shown in Table 2, the body weight of mice in each group increased in the experiment. After the modeling was completed, the body weight of the remaining four groups was significantly decreased compared with the normal group ( p<0.05 ). Compared with the model group, the body weight of the positive control group was significantly decreased ( p<0.05 ), while the heat-inactivated L-AKK IMMU group and H-AKK IMMU group had no significant change ( p>0.05 ), and there was no obvious difference between the two.

[0074] Pathological observation of detection example 1 After the fixed nose tissue was embedded in paraffin and sectioned, hematoxylin and eosin staining was used for observing pathological changes under a BX43 microscope (Olympus, Tokyo, Japan).

[0075] The HE staining results of mouse nose tissue showed that ( Figure 1 ) the nasal mucosa epithelial structure of the normal group of mice was complete and arranged in order without hypertrophy and hyperplasia, no interstitial edema and vascular dilation was observed, and only a small amount of lymphocytes and neutrophil infiltration was observed; the nasal mucosa epithelial cells of the model group of mice were hyperplastic and hypertrophic, the tissue structure was arranged in disorder, the mucosa epithelium was eroded, the intercellular space was edematous, and the blood vessels were dilated, and inflammatory cells such as eosinophils infiltrated; after being given the positive control group and heat-inactivated L-AKK IMMU and H-AKK IMMU pretreatment, the mouse nasal mucosa tissue structure was still present, the interstitial edema and small blood vessel dilation were significantly reduced compared with the model group, and the eosinophil infiltration was less, of which the AKK IMMU treatment group reduced the above damage in a dose-dependent manner, and the high-dose was similar to the montelukast group.

[0076] Measurement of serum oxidative damage-related biochemical indicators in detection example 2 Serum preparation: the mouse blood was placed at 4°C for 2 hours, centrifuged at 3000 rpm and 4°C for 15 minutes, and then the upper serum was obtained and frozen at -80°C. The oxidative stress indicators in the serum were detected: the levels of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and nitric oxide (NO) were determined by a kit to evaluate the oxidative damage related to allergic reactions.

[0077] MDA, NO, GSH, SOD, CAT can reflect the body's oxidative damage and antioxidant capacity. As shown in Table 2, compared with the normal group, the model group serum MDA, NO content increased, GSH, SOD, CAT enzyme decreased, the difference was statistically significant (P <0.05). Figure 2 p<0.05 Compared with the model group, after pretreatment with loratadine, heat-inactivated L-AKK IMMU, H-AKK IMMU group, serum MDA, NO decreased, GSH, SOD, CAT increased, the difference was statistically significant (P <0.05). Among them, in terms of reducing MDA and NO, heat-inactivated H-AKK IMMU group and loratadine showed similar performance, both stronger than heat-inactivated L-AKK IMMU group; In terms of improving GSH, SOD and CAT, heat-inactivated H-AKK IMMU group performed best, higher than loratadine and heat-inactivated L-AKK IMMU group, the latter was similar. p<0.05

[0078] Detection Example 3: Determination of cytokines related to inflammatory damage of lung tissue Preparation of lung tissue homogenate: 0.1 g of mouse lung tissue was added to 0.9 mL of physiological saline, and the tissue was homogenized at a speed of 6 m / s (Bioprep-24; Hangzhou Auson Instruments Co., Ltd., Hangzhou, China) for 5 times, 30 seconds each time. Index detection: The levels of cytokines interleukin IL-1β, IL-6, IL-10, IL-17A and IFN-γ in lung tissue were determined by enzyme-linked immunosorbent assay kit.

[0079] The inflammatory indicators of lung tissue were detected. As shown in Table 3, compared with the normal group, the expression of IL-1β, -6, 17A, -10 in the model group increased, while the expression of IFN-γ decreased. Figure 3 p<0.05 Compared with the model group, pretreatment with loratadine, heat-inactivated L-AKK IMMU, H-AKK IMMU group had a down-regulating effect on IL-1β, IL-6, IL-10, IL-17A, and the effect was loratadine ≥ H-AKK IMMU > L-AKK IMMU, but had an up-regulating effect on IFN-γ, and the three showed opposite trends.

[0080] Detection Example 4: Determination of cytokines related to immune function of spleen tissue Preparation of spleen tissue homogenate: 0.1 g of mouse spleen tissue was added to 0.9 mL of physiological saline, and the tissue was homogenized at a speed of 6 m / s (Bioprep-24; Hangzhou Auson Instruments Co., Ltd., Hangzhou, China) for 4 times, 30 seconds each time. Spleen OVA specific antibody level detection: The levels of spleen immunoglobulin IgG and IgM were detected by ELISA method. ​​​

[0081] As shown in Figure 4 , compared with the normal group, the OVA-specific IgM and IgG levels in the spleen tissue of the model group mice increased ( p< 0.05 ). Compared with the model group, the OVA-specific IgM and IgG levels in the spleen tissue of the loratadine, heat-inactivated L-AKK IMMU, and H-AKK IMMU groups decreased ( p<0.05 ), but the differences in IgM and IgG levels among the three groups were not statistically significant ( p>0.05 ).

[0082] Detection Example 5 After homogenizing the lung tissue, total RNA was extracted using TRIzol reagent, and the concentration was diluted to 1 μg / μL. The total RNA was reverse transcribed into cDNA. Then, 1.0 μL of cDNA, 2.0 μL of forward and reverse primers (10 μm), 10.0 μL of SYBR premix, and 7.0 μL of sterile ultrapure water were mixed, and amplification was performed under the following conditions: pre-denaturation at 95°C for 5 minutes; amplification reaction at 95°C for 10 seconds and at 60°C for 30 seconds, for 40 cycles. β-actin was the internal reference gene, and the data were analyzed by 2 - Ct Method calculation. The primer sequences are shown in Table 3.

[0083] Table 3 Primer sequences

[0084] The expression of IL-1β, IL-4, IL-6, IL-10, IL-17A, and IFN-γ genes in the lung tissue of mice was analyzed. The results of real-time fluorescent quantitative PCR are shown in Figure 5 Compared with the normal group, the expression of IL-1β, IL-4, IL-6, IL-10, IL-17A, and IFN-γ genes in the model group increased significantly, while IFN-γ decreased ( p<0.05 ). After intervention by loratadine, heat-inactivated L-AKK IMMU, and H-AKK IMMU groups, the expression of IL-1β, IL-4, IL-6, IL-10, and IL-17A genes decreased, and the expression of IFN-γ increased. Among them, loratadine and heat-inactivated H-AKK IMMU groups had similar effects and were close to the normal group, while the heat-inactivated L-AKK IMMU group had weaker anti-inflammatory ability and was inferior to the first two groups; the anti-inflammatory effect of the AKK IMMU group showed a dose-dependent manner.

[0085] Finally, it should be noted that the above is only to illustrate the technical solutions of the present application, and is not a limitation on the scope of protection of the present application, and simple modifications or equivalent replacements of the technical solutions of the present application by those skilled in the art do not deviate from the essence and scope of the technical solutions of the present application.

Claims

1. A strain of Akkermansia muciniphila (AkkIMMU), characterized in that, Akkermansia muciniphila ) AKK IMMU, characterized in that, The Akkermansia muciniphila AKK IMMU was deposited with the Guangdong Microbial Culture Collection Center on November 3, 2025, and the deposit number is GDMCC No: 67192.

2. Use of the Akkermansia muciniphila AKK IMMU of claim 1 in the preparation of a product for improving immunity.

3. Use of the Akkermansia muciniphila AKK IMMU of claim 1 in the preparation of a product for prophylactic treatment and / or adjuvant treatment of rhinitis.

4. Use according to claim 3, characterized in that, The prophylactic treatment and / or adjuvant treatment of rhinitis is achieved by at least one of the following mechanisms: (1) inhibiting the synthesis and release of inflammatory factors, reducing inflammatory damage to the mucosa, and repairing the epithelial structure; (2) improving the function of the antioxidant system, reducing the stimulation of oxidative damage to the nasal mucosa; (3) inhibiting specific immune responses, reducing antibody production, and reducing allergic reactions.

5. A product for boosting immunity and / or prophylactically treating and / or adjuvantly treating rhinitis, characterized in that, The product comprises the Akkermansia muciniphila AKK IMMU of claim 1.

6. The product of claim 5, wherein, The product comprises at least one of the culture solution, whole bacteria, or fermentation broth of the Akkermansia muciniphila AKK IMMU of claim 1.

7. The product of claim 5, wherein, The product comprises food and drugs.

8. The product of claim 7, wherein, The application amount of the product is not less than 10 CFU / kg according to the number of colonies. 7 CFU / kg.

9. The product of claim 8, wherein, The application rate of the product is 10 7 -10 9 CFU / kg.

10. The product of claim 5, wherein, The product further comprises excipients, which include any one or more of binding agents, fillers, disintegrating agents, lubricants, preservatives, antioxidants, flavorings, fragrances, solubilizers, emulsifiers, solubilizers, or osmotic pressure regulators. The product further comprises excipients, which include any one or more of binding agents, fillers, disintegrating agents, lubricants, preservatives, antioxidants, flavorings, fragrances, solubilizers, emulsifiers, solubilizers, or osmotic pressure regulators.

Images