Application of AKK bacteria in improving immunity, preparing prophylactic and therapeutic products for rhinitis and / or adjuvant therapy

Abstract

This invention, entitled "Application of Akkermansia myxophilus in Enhancing Immunity and Preparing Products for the Prevention, Treatment, and / or Adjunctive Treatment of Rhinitis," belongs to the field of microbial technology. Existing treatments for rhinitis suffer from problems such as short-lasting symptom relief, limited single-effect anti-inflammatory properties, and a tendency to cause side effects. To address these issues, this invention provides a novel Akkermansia myxophilus strain, AKK IMMU, which was deposited on November 3, 2025, at the Guangdong Provincial Center for Microbial Culture Collection (GDMCC No: 67192). This strain significantly improves nasal mucosal pathological damage in rhinitis model mice, regulates serum oxidative stress indicators, and modulates inflammatory factors in lung tissue.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to the application of AKK bacteria in the preparation of products for the prevention, treatment, and / or adjunctive treatment of rhinitis to enhance immunity. Background Technology

[0002] Rhinitis is an extremely 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 the immune system. It not only causes local symptoms such as nasal congestion, runny nose, sneezing, and nasal itching, but can also lead to sleep disorders, difficulty concentrating, and other problems, severely interfering with patients' daily lives, work, and studies. More seriously, allergic rhinitis is closely related to other allergic diseases such as asthma, significantly increasing the risk of developing asthma, posing a dual threat to patients' physical and mental health. Recent studies have shown that mouse models have important value in the study of rhinitis mechanisms. They can not only reflect the pathological characteristics of human rhinitis but also be used to evaluate the effectiveness of novel treatments. Analysis of the immune response, pathological changes, and inflammatory factor levels in mice can provide a deeper understanding of the occurrence and development of rhinitis.

[0003] Currently, the treatment of allergic rhinitis mainly relies on medications, such as antihistamines and corticosteroids. However, these medications can only relieve symptoms and cannot cure the disease at its root. Furthermore, long-term use can lead to serious side effects, such as hormone dependence and drug resistance, posing potential risks to patients' health. In addition, while desensitization therapy is a potential cure, its long treatment course, high cost, and ineffectiveness for all patients limit its widespread application. Therefore, developing safe and effective new treatment methods or interventions has become an urgent need in the field of allergic rhinitis prevention and treatment.

[0004] In recent years, probiotics have shown great potential as a novel biological intervention in the prevention and treatment of allergic diseases. Probiotics are a class of live microorganisms that are beneficial to the host, capable of regulating the intestinal microecological balance, enhancing intestinal barrier function, modulating the immune system, and reducing inflammatory responses. Multiple studies have shown that specific probiotic strains can alleviate the symptoms of allergic diseases by regulating the activity of immune cells and inhibiting the production of allergy-related cytokines.

[0005] Relevant patent documents retrieved:

[0006] The publication, published in China (CN114514028A) on May 17, 2022, discloses a pharmaceutical composition containing Akkermansia myxophilus EBAMDK19 strain or its culture or dried form for the effective prevention or treatment of atopic diseases. This pharmaceutical composition is capable of producing the same level of prevention or treatment effect as steroidal drugs for atopic diseases such as asthma, atopic dermatitis, urticaria, allergic rhinitis, allergic reactions, or food allergies.

[0007] The prior art represented by the aforementioned documents has at least the following unresolved technical problems or defects:

[0008] Existing technologies only offer general treatment options for panatopic diseases using *Akermansia myxophilus*, with allergic rhinitis merely listed as one of the atopic diseases. No technical solutions have been designed or experimentally validated specifically for the pathological mechanisms, core symptoms, local drug delivery requirements, and dedicated testing indicators of rhinitis. Summary of the Invention

[0009] The purpose of this invention is to provide:

[0010] The application of AKK bacteria in enhancing immunity, preparing products for the prevention, treatment and / or adjuvant treatment of rhinitis, and related technologies, to solve the technical problems of existing rhinitis treatments, such as unsustainable symptom relief, limited single anti-inflammatory effects, and easy to cause side effects, or combinations thereof.

[0011] Terminology Explanation:

[0012] Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0013] 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.

[0014] The definition of standard chemical terminology can be found in the reference "Microbiology", edited by Shen Ping and Chen Xiangdong, Higher Education Press.

[0015] Unless otherwise specified, conventional methods within the scope of the art should be used, such as ELISA kits for detecting inflammatory factors and neurotransmitters, RT-qPCR for detecting relative gene expression levels, and histopathological HE staining observation.

[0016] 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.

[0017] The terms "optional / arbitrary" or "optionally / arbitrarily" refer to events or situations described subsequently that may or may not occur, including both the occurrence and non-occurrence of the event or situation. For example, "optionally added excipients" means "no excipients added" or "excipients added".

[0018] The term "heat-inactivated AKK IMMU" as used in this article refers to the strain obtained by thawing the AKK IMMU strain from a -80℃ cryopreservation tube, followed by resuscitation culture in MRS liquid medium (37℃, 24h), expansion culture (37℃, 18h, 3% inoculum), centrifugation (4℃, 4000rpm, 10min) to collect the strain, and then heat inactivation treatment at 80℃ for 30min.

[0019] The term "rhinitis animal model" used in this article refers to a C57BL / 6 mouse rhinitis model constructed by intraperitoneal injection of ovalbumin (OVA) for basal sensitization (0.1 mg / mL OVA + 2 mg / mL aluminum hydroxide mixed solution, 0.2 mL / time, every other day, for a total of 7 times, 1-13 days) and OVA nasal challenge (5% OVA saline solution, 10 μL / nostril, once a day, for a total of 7 times, 15-21 days), and the model mice having a total nasal symptom score of more than 5 points.

[0020] The term "oxidative stress index" used in this article refers to indicators that can reflect the body's oxidative damage and antioxidant capacity, including malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and nitric oxide (NO).

[0021] The term “inflammatory factors” used in this article refers to cytokines involved in the inflammatory response, including interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-17A (IL-17A), and interferon-γ (IFN-γ).

[0022] The term "immunoglobulin" as used in this article refers to antibodies involved in immune responses, including immunoglobulin G (IgG) and immunoglobulin M (IgM).

[0023] In a first aspect, the present invention provides: a strain of Akkermansia myxophilus AKK IMMU, which was deposited on November 3, 2025, at the Guangdong Provincial Center for Microbial Culture Collection, with accession number GDMCC No:67192.

[0024] Secondly, the present invention provides the application of the above-mentioned Akkermansia AKK IMMU, a myxotroph, in the preparation of products that enhance immunity.

[0025] Thirdly, the present invention provides the use of the above-mentioned Akkermansia myxophilus AKK IMMU in the preparation of products for the treatment and / or adjunctive treatment of rhinitis.

[0026] The treatment and / or adjunctive treatment of rhinitis are achieved through at least one of the following mechanisms:

[0027] (1) Inhibits the synthesis and release of inflammatory factors, reduces mucosal inflammatory damage, and repairs epithelial structure;

[0028] (2) Enhance the function of the antioxidant system and reduce the irritation of oxidative damage to the nasal mucosa;

[0029] (3) Suppress specific immune responses, reduce antibody production, and alleviate allergic reactions.

[0030] Fourthly, the present invention provides a product comprising the above-mentioned Akkermansia AKK IMMU, a myxotrophic bacterium.

[0031] The product includes at least one of the following: culture medium, whole culture, or fermentation broth of Akkermansia AKK IMMU, a myxotrophic bacterium.

[0032] The products mentioned include, but are not limited to, food and pharmaceuticals.

[0033] The food products mentioned include, but are not limited to: health products, beverages, dairy products, solid snacks, and fermented foods.

[0034] The health products mentioned include, but are not limited to: probiotic capsules, probiotic powders, and probiotic tablets.

[0035] The beverages mentioned include, but are not limited to: fermented milk, lactic acid bacteria beverages, and probiotic oral liquids.

[0036] The dairy products include, but are not limited to: fermented milk, probiotic cheese, and probiotic milk powder; the solid snacks include probiotic biscuits, probiotic oatmeal, and probiotic pastries.

[0037] The fermented foods include, but are not limited to: probiotic fermented soy products and probiotic fermented grain products.

[0038] The drugs mentioned include, but are not limited to: probiotic preparations and compound preparations containing probiotics.

[0039] The application rate of the product, based on the number of colonies, is not less than 10. 7 CFU / kg.

[0040] The preferred application rate of the product, based on colony count, is 1×10⁻⁶. 7 -1×10 9 CFU / kg.

[0041] The dosage forms of the product include, but are not limited to: liquid dosage forms, gas dosage forms, solid dosage forms, or semi-solid dosage forms.

[0042] The liquid dosage forms include: solvent-based, aromatic aqueous solutions, tinctures, elixirs, colloidal solutions, pastes, suspensions, or emulsifiers.

[0043] The gaseous dosage form includes: aerosol or spray.

[0044] The solid dosage forms include: powders, pills, tablets, or films.

[0045] The semi-solid dosage forms include ointments, suppositories, or pastes.

[0046] The preferred dosage form is at least one of the following: liquid solution, lyophilized powder, tablet, capsule, granule, spray, oral dispersible tablet / film, and sublingual tablet / lozenge.

[0047] The product also includes auxiliary materials.

[0048] The excipients include, but are not limited to, any one or more of the following: binders, fillers, disintegrants, lubricants, preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solubilizers, or osmotic pressure regulators.

[0049] The excipients are preferably at least one of lactose, mannitol, methylcellulose, benzoic acid, sorbic acid, food coloring and / or talc.

[0050] 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 third aspect of the present invention includes:

[0051] The application of Akkermansia myxophilus AKK IMMU in the preparation of products for alleviating low-calcium diet-induced osteoporosis, wherein the dosage of Akkermansia myxophilus AKK IMMU in the product is 1×10⁻⁶. 7 CFU / kg - 1×10 9 The dosage form is CFU / kg, and the formulation is a bacterial liquid. This technical solution not only solves the technical problem of "alleviating osteoporosis caused by a low-calcium diet", but also further solves the technical problems of "product ease of use and dosage accuracy".

[0052] The present invention has at least the following beneficial effects:

[0053] 1. Compared with existing technologies (such as loratadine treatment), the present invention has better technical effects in improving nasal mucosal pathological damage, regulating serum oxidative stress indicators, and regulating lung tissue inflammatory factors in rhinitis model mice.

[0054] According to experimental tests, this invention (heat-inactivated AKK IMMU group) reduced the nasal symptom score of rhinitis model mice from 6.67±0.82 in the model group to below 4.33±0.52; reduced the serum MDA content from the elevated level in the model group to near the normal group (the baseline MDA level in the normal group is 0.83±0.41); reduced the lung tissue IL-1β content from the elevated level in the model group to near the normal group (the baseline IL-1β level in the normal group is 0.83±0.41); and reduced the degree of nasal mucosal epithelial hyperplasia and edema from "significant hyperplasia and severe edema" in the model group to below "mild hyperplasia and mild edema".

[0055] Considering the possibility of this invention entering other countries, this invention also provides the following technical solutions:

[0056] The present invention provides a method for the prevention, treatment and / or adjunctive treatment of rhinitis, comprising administering to a subject a therapeutically effective amount of Akkermansia myxophilus AKK IMMU.

[0057] The term "subject" includes living organisms (e.g., mammals) that can elicit an immune response. Examples of subjects include humans, primates, cattle, horses, goats, sheep, dogs, cats, mice, rats, rabbits, guinea pigs, pigs, and their transgenic species.

[0058] The terms “treat” or “treatment” or “ameliorate” refer to the medical management of a subject’s disease, condition, or undesirable condition. Treatment or preventative benefits include improved clinical outcomes; reduction or alleviation of symptoms associated with the disease, condition, or undesirable condition; reduced symptom occurrence; improved quality of life; longer disease-free status; reduction in the severity of the disease, condition, or undesirable condition; stabilization of the disease state; delay in disease progression; remission; survival; prolonged survival; or any combination thereof.

[0059] The term "therapeutic effective dose" refers to a pharmaceutically considered effective dosage, that is, an amount of active drug sufficient to significantly improve the condition without causing serious side effects. Dosage depends on many factors, such as the nature and severity of the disease to be prevented or treated, the sex, age, weight, personality, and individual response of the patient or animal, the route of administration, frequency of administration, and therapeutic purpose; therefore, the dosage of this invention can vary widely.

[0060] Preservation Instructions

[0061] Preserved strains: Akkermansia muciniphila AKK IMMU;

[0062] Category Naming: Akkermansia muciniphila ;

[0063] Accession number: GDMCC No:67192;

[0064] Preservation period: November 3, 2025;

[0065] Preservation institution: Guangdong Provincial Center for Microbial Culture Collection;

[0066] Abbreviation of depositary institution: GDMCC;

[0067] Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Attached Figure Description

[0068] Figure 1 The image shows pathological changes in mouse nasal tissue.

[0069] Figure 2 The values ​​shown in the figure represent the serum oxidative damage indicators of mice: MDA, NO, GSH, CAT, and SOD. The values ​​in the figure are the mean ± standard deviation. According to the Ducan test, there are significant differences in the mean values ​​of different letters within the same figure. p<0.05 ).

[0070] Figure 3 Inflammation markers in mouse lung tissue IL-1β, IL-6, IL-10, IL-17A and IFN-γThe values ​​shown in the figure are the mean ± standard deviation. According to the Ducan test, there are significant differences in the mean values ​​of different letters in the same figure. p< 0.05 ).

[0071] Figure 4 The values ​​shown in ac represent the levels of IgG and IgM, immune markers in mouse spleen tissue. The values ​​are the mean ± standard deviation. According to Ducan's test, there are significant differences in the mean values ​​of different letters within the same graph. p<0.05 ).

[0072] Figure 5 The values ​​shown in the figure represent the gene expression of IL-1β, IL-4, IL-6, IL-10, IL-17A, IFN-γ, and IFN-γ in mouse brain tissue. The values ​​in the figure are mean ± standard deviation. According to the Ducan test, there are significant differences in the mean values ​​of different letters within the same figure. p<0.05 ). Detailed Implementation

[0073] Unless otherwise specified, all raw materials and reagents used in this invention were purchased from commercial suppliers, and experiments were conducted in accordance with the operating instructions. Unless otherwise specified, all instruments, equipment, and apparatus used in this invention are conventional instruments, equipment, and apparatus, and experiments were conducted in accordance with the operating instructions and the accompanying reagents.

[0074] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. Unless otherwise specified in the embodiments, conditions are performed under conventional conditions or conditions recommended by the manufacturer. All reagents or instruments without specified manufacturers are commercially available conventional products. Numerous specific details are provided in the following detailed embodiments to better illustrate the invention. The specific embodiments described herein are for illustrative purposes only and are not intended to constitute any limitation on the invention.

[0075] All experimental data were statistically analyzed using statistical software, combined with one-way ANOVA and Duncan's method for multiple comparisons; p＜0.05 Data are considered statistically significant. Data are presented as mean ± standard deviation (SD), repeated 3 times.

[0076] Table 1. Reagents and their sources

[0077]

[0078] Basic Example 1: Preparation of Bacterial Culture

[0079] Take out the experimental strain cryopreservation tube of strain AKK IMMU from -80 °C. After thawing, inoculate (200 μL) into MRS liquid medium (8 mL), resuscitate and culture at 37 °C for 24 hours, then mix well and transfer 3% inoculum to MRS liquid medium and continue to culture for 18 hours. Centrifuge (4 °C, 4000 rpm, 10 minutes) to collect the AKK IMMU strain at the bottom, and resuspend it with physiological saline to adjust the cell concentration to 1.0×10 9 CFU / kg·bw and 1.0×10 7 CFU / kg·bw order of magnitude.

[0080] Basic Example 2 Reagent Preparation

[0081] Preparation of intraperitoneal injection solution: Weigh 1 mg of OVA powder and 20 mg of aluminum hydroxide powder in a disinfected environment in a laminar flow hood, add 10 ml of physiological saline, and mix well and shake. Prepare a final concentration of 0.1 mg / mL OVA + 2 mg / mL aluminum hydroxide mixed suspension for intraperitoneal injection in the basic sensitization stage.

[0082] Preparation of nasal challenge solution: Weigh 50 mg of OVA dry powder in a disinfected environment in a laminar flow hood and place it in a 3 mL sterile vial, add 1 mL of sterile physiological saline, mix well and shake, and prepare a 5% OVA physiological saline solution with a final concentration for nasal challenge.

[0083] Example 1 Animal Model Construction

[0084] 30 C57BL / 6 mice, male, SPF grade, 6 - 8 weeks old, were purchased from Hunan Slack Jingda Experimental Animal Co., Ltd. Animal production license number: SYXK(Xiang)2024 - 0019. The mice were housed in standard mouse cages in the laboratory animal room, with a room temperature of 23 ± 2 °C and a relative humidity of 55 ± 5%. The lights were turned on for illumination every 12 hours, and they had free access to food and water and were adaptively raised for one week. The mice were randomly divided into 5 groups, with 6 mice in each group. After adaptive feeding, they were given drugs for model establishment. This study was approved by the Ethics Committee of the Collaborative Innovation Center for Child Nutrition and Health Development of Chongqing University of Education (2024112502B).

[0085] (1) Normal group: Without intervention.

[0086] (2) Model group: An AR animal model was established by intraperitoneal injection of OVA for basal sensitization and nasal instillation of OVA for nasal provocation. The specific operation method was as follows: from day 1 to 13, mice were injected intraperitoneally every other day with 0.2 mL of a mixed solution containing 0.1 mg / mL OVA + 2 mg / mL aluminum hydroxide for a total of seven times to basally sensitize them. Starting from day 15, 10 μL of 5% OVA solution was continuously instilled into the nostrils for a total of seven times to nasally provocation until day 21.

[0087] (3) Positive control group: An AR animal model was established by intraperitoneal injection of OVA for basal sensitization and nasal provocation by OVA drops. During the modeling period, loratadine (2 mg / kg) was administered by gavage for 3 weeks. The specific operation method was as follows: From day 1 to day 13, 0.2 ml of a mixed solution containing 0.1 mg / mL OVA + 2 mg / mL aluminum hydroxide was injected intraperitoneally every other day for a total of seven times to basally sensitize the mice. From day 15, 10 μL of 5% OVA solution was continuously dripped into the nostrils daily for a total of seven times to nasal provocation until day 21. At the same time, loratadine (2 mg / kg) was administered by gavage every other day from day 1 to day 21.

[0088] (4) Heat-inactivated AKK IMMU low-dose group (1.0×10 7 CFU / kg·bw): An AR animal model was established using intraperitoneal injection of OVA for basal sensitization and nasal provocation via OVA nasal drops. During the modeling period, mice were administered the probiotic AKK IMMU via gavage for 3 weeks. Specifically, from days 1 to 13, mice were basally sensitized by intraperitoneal injection of 0.2 ml of a mixed solution containing 0.1 mg / mL OVA and 2 mg / mL aluminum hydroxide every other day for a total of seven times. From day 15, nasal provocation was performed by intranasal drops of 10 μL of 5% OVA solution into the nostrils seven times daily until day 21. Simultaneously, from day 1 to day 21, mice were given 1.0 × 10⁻⁶ heat-inactivated (80℃, 30 min) daily. 7 AKK IMMU administered via gavage at the level of CFU / kg·bw.

[0089] (5) High-dose group of heat-inactivated AKK IMMU (1.0×10 9CFU / kg·bw): An AR animal model was established using intraperitoneal injection of OVA for basal sensitization and nasal challenge with OVA intranasal drops. During the modeling period, mice were administered the probiotic AKK IMMU by gavage for 3 weeks. Specifically, from days 1 to 13, mice were basally sensitized by intraperitoneal injection of 0.2 ml of a mixed solution containing 0.1 mg / mL OVA and 2 mg / mL aluminum hydroxide every other day for a total of seven times. From day 15, nasal challenge was performed by intranasal drops of 10 μL of 5% OVA solution into the nostrils seven times daily until day 21. Simultaneously, from day 1 to day 21, mice were given 1.0 × 10⁻⁶ heat-inactivated (80℃, 30 min) daily. 9 AKK IMMU administered via gavage at the level of CFU / kg·bw.

[0090] After the last nasal stimulation, each mouse was placed in an independent cage for 30 minutes to observe the occurrence and severity of symptoms such as nose scratching, sneezing, and runny nose. The mice were scored according to their nasal symptoms. The scoring criteria were as follows: (1) Nasal itching: 1 point for lightly rubbing the nose a few times, 2 points for repeatedly scratching the nose and face, and 3 points for rubbing the nose and face all over. (2) Sneezing: 1-3 sneezes, 2 points for 4-10 sneezes, and 3 points for more than 11 sneezes. (3) Clear nasal discharge: 1 point for discharge flowing into the nostrils, 2 points for discharge flowing beyond the nostrils, and 3 points for discharge flowing onto the face. The scores of each nasal symptom were recorded using the superposition quantification method. Then, the total symptom score for each mouse was calculated. A total score of more than 5 points indicates that the AR model has been successfully established and can be used for subsequent experiments.

[0091] After weighing, the mice were anesthetized with ether, their eyeballs were enucleated, and blood was collected. They were then euthanized by cervical dislocation. Nasal hair was removed, the nose was cut off, decalcified, and fixed in fixative for at least 24 hours. Lung and spleen tissues were obtained from dissection and stored at -80°C for subsequent experiments.

[0092] The results of clinical symptom observation and weight change in mice are shown in Table 2.

[0093] Table 2 Clinical symptom observation and weight changes

[0094]

[0095] a-d The values ​​shown are the mean ± standard deviation. According to the Ducan test, there are significant differences in the mean values ​​of different letters in the same column. p<0.05 ).

[0096] Compared with the normal group, all model group mice had symptom scores ≥5, indicating that the AR model was successfully established in this study, and the differences were statistically significant. p<0.05Compared with the model group mice, the symptom scores of the positive control group, the heat-inactivated L-AKK IMMU group, and the H-AKK IMMU group were reduced. p<0.05 Among them, the positive control group showed more significant effects, while the symptom scores of the H-AKK IMMU group were not significantly different from those of the L-AKK IMMU group. p>0.05 ).

[0097] As shown in Table 2, the body weight of mice in all groups increased during the experiment. After the modeling was completed, the body weight of the other four groups decreased significantly compared with the normal group. p<0.05 Compared with the model group mice, the positive control group mice showed a significant decrease ( ). p<0.05 ), while no significant changes were observed in the heat-inactivated L-AKK IMMU group and H-AKK IMMU group (). p>0.05 There is no significant difference between the two.

[0098] Pathological observation of case 1

[0099] After the fixed nasal tissue was embedded in paraffin and sectioned, it was stained with hematoxylin and eosin for observation of pathological changes under a BX43 microscope (Olympus, Tokyo, Japan).

[0100] HE staining results of mouse nasal tissue showed ( Figure 1 In the normal group, the nasal mucosal epithelial structure of mice was intact and neatly arranged without hypertrophy or proliferation, and no interstitial edema or vasodilation was observed, with only a small number of lymphocytes and neutrophils infiltrating. In the model group, the nasal mucosal epithelial cells of mice showed hyperplasia and hypertrophy, disordered tissue structure, mucosal epithelial erosion, intercellular edema and vasodilation, and infiltration of inflammatory cells such as eosinophils. After pretreatment with the positive control group and heat-inactivated L-AKK IMMU and H-AKK IMMU, the nasal mucosal tissue structure of mice was still intact, and the interstitial edema and small blood vessel dilation were significantly reduced compared with the model group, and the eosinophil infiltration was reduced. Among them, the AKK IMMU treatment group reduced the above-mentioned damage in a dose-dependent manner, and the high dose was similar to that of the loratadine group.

[0101] Example 2: Determination of serum oxidative damage-related biochemical indicators

[0102] Serum preparation: Mouse blood was incubated at 4°C for 2 hours, centrifuged at 3000 rpm and 4°C for 15 minutes, and the supernatant serum was obtained and frozen at -80°C. Detection of oxidative stress markers in serum: The levels of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), and nitric oxide (NO) were measured using a kit to assess oxidative damage associated with allergic reactions.

[0103] MDA, NO, GSH, SOD, and CAT can reflect the body's oxidative damage and antioxidant capacity. For example... Figure 2 As shown, compared with the normal group, the serum levels of MDA and NO in the model group were increased, while the levels of GSH, SOD, and CAT enzymes were decreased, and the differences were statistically significant. p<0.05 Compared with the model group, pretreatment with loratadine, heat-inactivated L-AKK IMMU, and H-AKK IMMU resulted in a decrease in serum MDA and NO, and an increase in GSH, SOD, and CAT, with statistically significant differences. p<0.05 Among them, in terms of reducing MDA and NO, the heat-inactivated H-AKK IMMU group and loratadine showed similar performance, both being stronger than the heat-inactivated L-AKK IMMU group; in terms of increasing GSH, SOD, and CAT, the heat-inactivated H-AKK IMMU group performed the best, higher than loratadine and the heat-inactivated L-AKK IMMU group, which were similar.

[0104] Example 3: Measurement of cytokines related to lung tissue inflammation and injury

[0105] Lung tissue homogenate preparation: 0.1 g of mouse lung tissue was added to 0.9 mL of physiological saline, and the tissue was homogenized 5 times at a rate of 6 m / s (Bioprep-24; Hangzhou Ausen Instruments Co., Ltd., Hangzhou, China) for 30 seconds each time. Indicator detection: The levels of cytokines interleukin IL-1β, IL-6, IL-10, IL-17A, and IFN-γ in the lung tissue were measured using an enzyme-linked immunosorbent assay (ELISA) kit.

[0106] Inflammatory markers in lung tissue are detected. For example... Figure 3 As shown, compared with the normal group, the expression of IL-1β, -6, 17A, and -10 increased in the model group, while the expression of IFN-γ decreased. p<0.05 Compared with the model group, pretreatment with loratadine, heat-inactivated L-AKK IMMU, and H-AKK IMMU downregulated IL-1β, IL-6, IL-10, and IL-17A, with the effect being loratadine ≥ H-AKK IMMU > L-AKK IMMU. However, it upregulated IFN-γ, and the three showed opposite trends.

[0107] Example 4: Assay of immune-related cytokines in spleen tissue

[0108] Spleen tissue homogenate preparation: 0.1 g of mouse spleen tissue was added to 0.9 mL of physiological saline, and the tissue was homogenized 4 times at a rate of 6 m / s (Bioprep-24; Hangzhou Ausen Instruments Co., Ltd., Hangzhou, China) for 30 seconds each time. Spleen OVA-specific antibody level detection: The levels of immunoglobulins IgG and IgM in spleen tissue were detected by ELISA.

[0109] like Figure 4 As shown, compared with the normal group, the levels of OVA-specific IgM and IgG in the spleen tissue of mice in the model group were increased ( p< 0.05 Compared with the model group, the levels of OVA-specific IgM and IgG in the spleen tissue of mice in the loratadine, heat-inactivated L-AKK IMMU, and H-AKK IMMU groups were decreased. p<0.05 However, there was no statistically significant difference in IgM and IgG levels among the three groups. p>0.05 ).

[0110] Case 5

[0111] After homogenization of lung tissue, total RNA was extracted using TRIzol reagent and 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 amplified under the following conditions: pre-denaturation at 95°C for 5 minutes; amplification reaction at 95°C for 10 seconds, followed by a 30-second reaction at 60°C, for 40 cycles. β-actin was used as an internal reference gene; data were obtained through 2... - Ct Methods for calculation. Primer sequences are shown in Table 3.

[0112] Table 3 Primer Sequences

[0113]

[0114] The expression of IL-1β, IL-4, IL-6, IL-10, IL-17A, and IFN-γ genes in mouse lung tissue was analyzed. Real-time quantitative PCR results are shown below. Figure 5 As shown, 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 all showed a significant increase, while the expression of IFN-γ decreased. p<0.05 After intervention with loratadine, heat-inactivated L-AKK IMMU, and H-AKK IMMU, the expression of IL-1β, IL-4, IL-6, IL-10, and IL-17A genes decreased, while the expression of IFN-γ increased. Among these, loratadine and heat-inactivated H-AKK IMMU showed similar effects, approaching those of the normal group, while the heat-inactivated L-AKK IMMU group exhibited weaker anti-inflammatory activity, ranking lower than the former two. The anti-inflammatory effect of the AKK IMMU group showed a dose-dependent relationship.

[0115] 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 strain of Akkermansia muciniphila AKK IMMU, characterized in that, The *Ackermania* AKK IMMU strain was deposited at the Guangdong Provincial Center for Microbial Culture Collection on November 3, 2025, with accession number GDMCC No:67192.

2. The use of the Ackeratophilic Akkermansia AKK IMMU as described in claim 1 in the preparation of a medicament for the prevention and / or adjunctive treatment of rhinitis.

3. The application according to claim 2, characterized in that, The prevention and / or adjunctive treatment of rhinitis are achieved through at least one of the following mechanisms: (1) Inhibits the synthesis and release of inflammatory factors, reduces mucosal inflammatory damage, and repairs epithelial structure; (2) Enhance the function of the antioxidant system and reduce the irritation of oxidative damage to the nasal mucosa; (3) Suppress specific immune responses, reduce antibody production, and alleviate allergic reactions.

4. A medicine for the prevention and / or adjunctive treatment of rhinitis, characterized in that, The drug comprises Akkermansia myxophilus AKK IMMU as described in claim 1, which has been heat-inactivated; the heat inactivation temperature is 80°C and the time is 30 min.

5. The pharmaceutical product according to claim 4, characterized in that, The amount of the drug product administered is not less than 10 CFU / kg in terms of the number of colonies. 7 CFU / kg.

6. The pharmaceutical product according to claim 4, characterized in that, The amount of the drug product administered is 10 7 -10 9 CFU / kg.

7. The pharmaceutical product according to claim 4, characterized in that, The drug also includes excipients, which include any one or more of the following: binders, fillers, disintegrants, lubricants, preservatives, antioxidants, flavoring agents, fragrances, solubilizers, emulsifiers, solubilizers, or osmotic pressure regulators.

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