Application of Akkermansia muciniphila and its postbiotics in improving respiratory diseases

Abstract

The present invention belongs to the field of microbial technology, and specifically relates to the application of Akkermansia muciniphila and its postbiotics in improving respiratory diseases. The Akkermansia muciniphila is AKK ONE, and the deposit number is CGMCC No.20954. AKK ONE of the present invention can effectively relieve cough caused by mycoplasma pneumonia, and down-regulate the content of pro-inflammatory factors IL-6 and IL-8 in mouse serum, and up-regulate the content of anti-inflammatory factor IL-10, thereby reducing the effect of inflammatory response; reducing the mRNA expression of tissue TRPA1, SP, and CGRP, and inhibiting the severe cough caused by inflammatory mycoplasma infection. The present invention provides a theoretical basis and experimental basis for the research and development of postbiotics and drugs for mycoplasma pneumonia.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to application of Akkermansia muciniphila and postbiotics in improving respiratory diseases. Background Art

[0002] Mycoplasma pneumonia is pneumonia caused by Mycoplasma pneumonia (MP). It has a slow onset, with fever, paroxysmal irritating cough, and a small amount of mucous or mucopurulent sputum (occasionally bloody sputum). Pulmonary signs are usually not obvious, but it can easily cause multi-system involvement outside the lungs, which can also threaten life or death.

[0003] Mycoplasma pneumoniae is a pathogen that once invades the human respiratory tract, the respiratory tract may cause partial damage and shedding of epithelial cells due to contact with pathogens, exposing the sensory nerve endings under the mucosa. [1] Then, through a series of transmitter transmission and release, cough sensitivity increases, which eventually leads to cough. Studies have shown that various transient receptor potentials (TRP) are expressed in the epithelium of airway sensory neurons. As important receptors on cough receptors, they play an important role in the pathogenesis of cough. [2] Transient receptor potential anchor protein 1 (TRPA1), also known as ANKTMI protein, is a member of the TRP family. [3] In mammals, TRPA1 is highly expressed in primary sensory neuron terminals and plays a role in the nociceptor-dependent perception of harmful stimuli in the environment. [4] TRPA1 is expressed on primary sensory neurons in the respiratory tract. When the respiratory tract is stimulated by external stimuli, intracellular Ca 2+ Increased concentrations lead to the release of substance P (SP) and calcitonin gene-related peptide (CGRP), which can cause bronchoconstriction and release inflammatory factors, leading to coughing. [5] Cough is the main symptom of Mycoplasma pneumoniae infection, but the commonly used cough suppressants do not achieve ideal therapeutic effects and have side effects during treatment.

[0004] Probiotics are active microorganisms that are beneficial to the human body. They are not antibiotics and are beneficial to the human body. They can improve the microecological balance of the human body, regulate the flora, kill bacteria in the human body, and have the effect of killing and inhibiting pathogenic bacteria such as mold, mycoplasma, and chlamydia. In addition, postbiotics are a general term for the metabolites of probiotics after processing, including bacteria and metabolites. Studies have confirmed that screened postbiotics are better than original live bacteria in enhancing immunity, and they still retain a high degree of physiological activity even after being exposed to high temperatures or treated with gastrointestinal digestive fluids.

[0005] The prior art does not disclose the use of Akkermansia muciniphila in treating mycoplasma pneumonia.

[0006] [1]Chaudhry R,Ghosh A,Chandolia A.Pathogenesis of Mycoplasmapneumoniae:An update[J].Indian J Med Microbiol.2016,34(1):7.

[0007] [2]Bonvini SJ,Belvisi MG. Cough and airway disease:The role of ionchannels[J].Pulm Pharmacol Ther.2017,47:21.

[0008] [3]Clapham DE, Montell C, Schultz G, et al. International Union of Pharmacology. International Union of Pharmacology.

[0009] [4] Li Fei, Chen Yuehan, Zhu Man. Research progress on the relationship between transient receptor potential channel ankyrin 1 and pain[J]. Chinese Journal of Geriatric Cardiovascular and Cerebrovascular Diseases, 2018, 20(11):1227.

[0010] [5]Yang H, Li S.Transient Receptor Potential Ankyrin 1(TRPA1)Channeland Neurogenic Inflammation in Pathogenesis of Asthma[J]. Med Sci Monit.2016,22:2917.

[0011] The present invention is the development of downstream application technology carried out by the inventor for the independently developed microbial resources: Akkermansia muciniphila with the deposit number of CGMCC No.20954 (other patents have been submitted on the same day).

[0012] After obtaining Akkermansia muciniphila, the inventors conducted application experiments in multiple fields including improving diabetes, anti-infection, improving respiratory diseases, regulating lipid metabolism, improving enteritis, etc., and obtained positive experimental results in some fields.

[0013] Taking into account the relevant provisions of the Patent Law on unity, protection is requested separately for each indication that does not have unity.

[0014] The present invention is directed to the protection of the application technology in improving respiratory diseases.

[0015] Since relevant evidence such as microbial identification and performance testing was published in the microbial patent submitted on the same day, in order to facilitate the review of the present invention and quickly understand the inventor's previous work, the relevant information of Akkermansiamuciniphila is briefly disclosed in the content of the present invention.

[0016] The conclusive information in this section is for the convenience of the review of the present invention, but is not intended to limit the scope of protection of the present invention. The specific experimental process and experimental results are recorded in the microbiological patent filed on the same day as the present invention.

[0017] 1. Akkermansia muciniphila was obtained by separation and screening by the present inventors.

[0018] 2. The deposit information of Akkermansia muciniphila: The deposit number is: CGMCC No.20954.

[0019] 3. Physiological and biochemical properties of Akkermansia muciniphila:

[0020] (1) Hydrophobicity: The hydrophobicity of the strain increased with time, reaching more than 30% at 60 min.

[0021] (2) Self-aggregation: The self-aggregation of the strain increased with time and tended to be stable at 20 hours, maintaining at around 50%.

[0022] (3) Gastrointestinal fluid tolerance:

[0023] The survival rate of the strain in gastric juice showed a downward trend with the extension of time, and the survival rate was about 80% at 240 minutes;

[0024] The survival rate of the strains in the intestinal fluid showed a downward trend as a whole, and the survival rate was above 80% at 240 minutes.

[0025] (4) Biofilm formation ability: weak.

[0026] (5 Bacterial reverse mutation test: The results of bacterial reverse mutation test showed that the strain was not mutagenic.

[0027] (6) Acute toxicity:

[0028] The mice were acutely gavaged with high, medium and low doses of AKK ONE, and no mortality occurred. The daily weight changes of the mice in the test groups were compared with those in the control saline group;

[0029] During the acute gavage period, there was no significant difference in the daily food intake of the test mice and the control saline group mice;

[0030] During the acute gavage period, there was no significant difference in the daily changes in blood glucose between the test mice and the control saline group mice;

[0031] There were no significant differences in the blood parameters between the acute gavage group and the control saline group;

[0032] There was no significant difference in triglyceride levels in serum and liver between the test mice and the control mice;

[0033] There was a very significant difference in liver cholesterol levels between mice given high doses orally and mice in the control group, but no significant differences were found in serum and liver in other groups.

[0034] There was no significant difference in the bile acid levels in serum and liver between the test mice and the control mice;

[0035] There was no significant difference in glucose levels in serum and liver between the test mice and the control mice;

[0036] There was no significant difference in the total protein levels in serum and liver between the test mice and the control mice;

[0037] There was no significant difference in the activity of alanine aminotransferase in serum and liver between the test mice and the control mice.

[0038] There was no significant difference in the activity of aspartate aminotransferase in serum and liver between the test mice and the control mice;

[0039] There was no significant difference in serum creatinine content between the test mice and the control mice;

[0040] There was no significant difference in the urea nitrogen concentration in the serum between the test mice and the control mice;

[0041] There was basically no significant difference in the weight of the heart, liver, spleen, kidney, thymus, brain, testis, lung, stomach and intestine of the test mice and the control mice;

[0042] There was no significant pathological damage in the liver and kidneys of the test mice and the control group mice.

[0043] (7) Subchronic toxicity test:

[0044] Mice were gavaged with high, medium and low doses of AKK ONE for 90 consecutive days without death. There was no significant difference in the weekly weight change between the test group and the control saline group;

[0045] During the continuous gavage period, there was no significant difference in the weekly changes in food intake between the test mice and the control saline group mice;

[0046] During the continuous gavage period, there was no significant difference in the blood glucose changes between the mice gavaged with a medium dose per week and the mice in the control saline group;

[0047] There were no significant differences in the blood parameters between the mice in the gavage group and the control saline group;

[0048] There was no significant difference in triglyceride levels in the liver and serum between the test mice and the control mice;

[0049] There was no significant difference in cholesterol levels in the liver and serum between the test mice and the control mice;

[0050] The bile acid levels in the liver of mice given medium doses of oral administration and those in the control group were significantly different, while there were no significant differences in the serum and liver between the other groups and the control group.

[0051] There was no significant difference in glucose levels in the liver and serum between the test mice and the control mice;

[0052] There was no significant difference in the total protein content in the liver and serum between the test mice and the control mice;

[0053] There was a significant difference in the activity of alanine aminotransferase in the serum of mice given the medium dose and the control group, but no significant difference was found in the serum and liver of the other groups.

[0054] There was a significant difference in the activity of alanine aminotransferase in the serum of mice given the medium dose and the control group, but no significant difference was found in the serum and liver of the other groups.

[0055] There was no significant difference in serum creatinine content between the test mice and the control mice;

[0056] There was no significant difference in the urea nitrogen concentration in the serum between the test mice and the control mice;

[0057] There was basically no significant difference in the weight of the heart, liver, spleen, kidney, thymus, brain, testis, lung, pancreas, stomach and intestine between the test mice and the control mice;

[0058] There was no significant pathological damage in the liver and kidneys of the test mice and the control mice;

[0059] There was no significant difference in blood sugar regulation ability between the test mice and the control group mice. Summary of the invention

[0060] In order to solve the above problems, the present invention provides a strain of Akkermansia muciniphila, named AKK ONE, which can be effectively used for the treatment of respiratory diseases, especially pneumonia caused by mycoplasma.

[0061] On the one hand, the present invention provides the use of Akkermansia muciniphila AKK ONE in the preparation of postbiotics for preventing and / or treating respiratory diseases, and the deposit number of the AKK ONE is CGMCC No.20954.

[0062] Specifically, the postbiotics include fermentation broth, fermentation broth supernatant, fermentation broth precipitate and / or dead bacteria of AKK ONE.

[0063] More specifically, the fermentation liquid refers to the liquid obtained by inoculating the bacteria into the culture medium and culturing it for a period of time.

[0064] To be more specific, the fermentation broth supernatant refers to the clarified liquid on the upper layer of the fermentation broth after centrifugation; it contains abundant metabolites of bacterial growth and reproduction and some bacterial fragments. The acidic substances and bacteriocins secreted by bacteria have antagonistic and killing effects on harmful bacteria; the amino acids and synthesized vitamins after bacteria decompose food are all in the culture medium, and also include enzymes secreted by bacteria that are useful to the human body; and some bacterial components also have an immune-promoting effect on the human body.

[0065] More specifically, the fermentation liquid precipitate refers to the liquid precipitate obtained by centrifugation, including free protein, residual bacteria, broken cells, and residues of the culture matrix, mainly protein and intracellular matrix.

[0066] More specifically, the dead bacteria are microorganisms that have lost their vitality and are unable to grow and reproduce. The probiotics lose their vitality due to the production process, such as high temperature treatment or excessive drying.

[0067] Specifically, the respiratory disease includes but is not limited to pneumonia.

[0068] More specifically, the pneumonia is mycoplasma pneumonia.

[0069] In another aspect, the present invention provides a postbiotic for preventing and / or treating respiratory diseases, wherein the postbiotic comprises AKK ONE; the deposit number of the AKK ONE is CGMCC No.20954.

[0070] Specifically, AKK ONE is its fermentation broth, fermentation broth supernatant, fermentation broth precipitate and / or dead bacteria.

[0071] More specifically, the fermentation liquid refers to the liquid obtained by inoculating the bacteria into the culture medium and culturing it for a period of time.

[0072] To be more specific, the fermentation broth supernatant refers to the clarified liquid on the upper layer of the fermentation broth after centrifugation; it contains abundant metabolites of bacterial growth and reproduction and some bacterial fragments. The acidic substances and bacteriocins secreted by bacteria have antagonistic and killing effects on harmful bacteria; the amino acids and synthesized vitamins after bacteria decompose food are all in the culture medium, and also include enzymes secreted by bacteria that are useful to the human body; and some bacterial components also have an immune-promoting effect on the human body.

[0073] More specifically, the fermentation liquid precipitate refers to the liquid precipitate obtained by centrifugation, including free protein, residual bacteria, broken cells, and residues of the culture matrix, mainly protein and intracellular matrix.

[0074] More specifically, the dead bacteria are microorganisms that have lost their vitality and are unable to grow and reproduce. The probiotics lose their vitality due to the production process, such as high temperature treatment or excessive drying.

[0075] Specifically, the respiratory disease includes but is not limited to pneumonia.

[0076] More specifically, the pneumonia is mycoplasma pneumonia.

[0077] In another aspect, the present invention provides a drug for preventing and / or treating respiratory diseases.

[0078] Specifically, the drug includes AKK ONE; the deposit number of AKK ONE is CGMCC No.20954.

[0079] More specifically, the drug includes fermentation broth, fermentation broth supernatant, fermentation broth precipitate, live bacteria, dead bacteria, lyophilized powder and / or cell lysate of AKK ONE.

[0080] More specifically, the fermentation liquid refers to the liquid obtained by inoculating the bacteria into the culture medium and culturing it for a period of time.

[0081] More specifically, the fermentation liquid supernatant refers to the clarified liquid on the upper layer of the fermentation liquid after centrifugation; it contains abundant metabolites of bacterial growth and reproduction and some bacterial fragments. The acidic substances and bacteriocins secreted by bacteria have antagonistic and killing effects on harmful bacteria; the amino acids and synthesized vitamins after bacteria decompose food are all in the culture medium, and also include enzymes secreted by bacteria that are useful to the human body; and some bacterial components also have an immune-promoting effect on the human body.

[0082] More specifically, the fermentation liquid precipitate refers to the liquid precipitate obtained by centrifugation, including free protein, residual bacteria, broken cells, and residues of the culture matrix, mainly protein and intracellular matrix.

[0083] More specifically, the live bacteria, also called active flora, can colonize and multiply in the intestine, which is beneficial to increase the number of beneficial bacteria.

[0084] More specifically, the dead bacteria are microorganisms that have lost their vitality and are unable to grow and reproduce. The probiotics lose their vitality due to the production process, such as high temperature treatment or excessive drying.

[0085] Specifically, the respiratory disease includes but is not limited to pneumonia.

[0086] More specifically, the pneumonia is mycoplasma pneumonia.

[0087] Specifically, the medicine also includes pharmaceutically acceptable excipients.

[0088] More specifically, the pharmaceutically acceptable excipient is selected from one or a combination of two or more of wetting agents, emulsifiers, preservatives, antioxidants, buffers, excipients, diluents, lubricants, antibacterial agents, suspending agents, suspending aids, solubilizers, thickeners, stabilizers, sweeteners and flavors.

[0089] Preferably, the pharmaceutically acceptable excipient is at least one selected from lactose, mannose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0090] In yet another aspect, the present invention provides a method for preparing postbiotics for preventing and / or treating respiratory diseases, the preparation method comprising culturing AKK ONE; the deposit number of the AKK ONE is CGMCC No.20954.

[0091] Specifically, the AKK ONE culture medium includes but is not limited to: BHI culture medium and MRS culture medium.

[0092] Specifically, the respiratory disease includes but is not limited to pneumonia.

[0093] More specifically, the pneumonia is mycoplasma pneumonia.

[0094] In another aspect, the present invention provides a method for preparing a drug for preventing and / or treating respiratory diseases, the preparation method comprising culturing AKK ONE; the deposit number of the AKK ONE is CGMCC No.20954.

[0095] Specifically, the AKK ONE culture medium includes but is not limited to: BHI culture medium and MRS culture medium.

[0096] More specifically, the concentration of AKK ONE is 1×10 8 TFU / mL-1×10 12 TFU / mL.

[0097] Preferably, the concentration of AKK ONE is 1×10 9 TFU / mL-1×10 10 TFU / mL.

[0098] Specifically, the respiratory disease includes but is not limited to pneumonia.

[0099] More specifically, the pneumonia is mycoplasma pneumonia.

[0100] Specifically, the medicine also includes pharmaceutically acceptable excipients.

[0101] More specifically, the pharmaceutically acceptable excipient is selected from one or a combination of two or more of wetting agents, emulsifiers, preservatives, antioxidants, buffers, excipients, diluents, lubricants, antibacterial agents, suspending agents, suspending aids, solubilizers, thickeners, stabilizers, sweeteners and flavors.

[0102] Preferably, the pharmaceutically acceptable excipient is at least one selected from lactose, mannose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil.

[0103] The technical effects achieved by the present invention are:

[0104] (1) AKK ONE can significantly reduce the frequency of coughing in mice.

[0105] (2) AKK ONE can slightly downregulate the levels of pro-inflammatory factors IL-6 and IL-8 in mouse serum, and upregulate the level of anti-inflammatory factor IL-10.

[0106] (3) The expression levels of TRPA1, SP, and CGRP mRNA in the lung tissue of mice intervened by AKK ONE were reduced.

[0107] Storage Instructions:

[0108] Strain name: AKK ONE;

[0109] Deposit number: CGMCC No.20954;

[0110] Taxonomic nomenclature: Akkermansia muciniphila;

[0111] Deposit date: October 26, 2020;

[0112] Depository: General Microbiology Center, China Microbiological Culture Collection Administration;

[0113] Address of the preservation unit: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0114] Figure 1 The number of coughs of mice within 5 minutes, where the same letters indicate no significant difference, and different letters indicate significant difference. DETAILED DESCRIPTION

[0115] The present invention will be further described in detail below in conjunction with specific examples. The following examples are not intended to limit the present invention, but are only intended to illustrate the present invention. The experimental methods used in the following examples are generally conventional, unless otherwise specified, and the materials, reagents, etc. used in the following examples are commercially available, unless otherwise specified.

[0116] the term:

[0117] Interleukin-6 (IL-6) is a typical pro-inflammatory cytokine secreted by monocytes, macrophages, activated T cells, B cells and fibroblasts.

[0118] Interleukin-10 (IL-10) has an anti-inflammatory effect. IL-10 can inhibit the production of inflammatory mediators and reduce the degree of inflammatory response; IL-10 can inhibit Th1 cell immune response and reduce cell-mediated immune inflammation; IL-10 can inhibit the activation of macrophages and dendritic cells and reduce the release of inflammation-related cytokines.

[0119] Interleukin-8 (IL-8) is an important chemokine and neutrophil activation factor and a pro-inflammatory factor.

[0120] Example 1

[0121] The strain of the present invention is Akkermansia muciniphila AKK ONE, whose deposit number is CGMCC No. 20954, hereinafter referred to as AKK ONE.

[0122] Preparation of AKK ONE dead bacteria resuspension: inoculate the frozen strain into BHI liquid culture medium, culture at 37℃ for 72h, centrifuge at 4℃ and 5000r / min for 5min, discard the supernatant, obtain the bacterial sludge, inoculate the bacterial sludge into BHI liquid culture medium at 3% inoculation amount, and activate at 37℃ for 72h. Use an inoculation loop to take a small amount of bacterial liquid and make three-zone lines on BHI solid culture medium, culture at 37℃ for 48h, pick a single colony and culture it in BHI liquid culture medium for 15h, and use it as a bacterial suspension after three consecutive activations. Count the live bacteria by flow cytometry, and the live bacteria concentration is 1×10 9 CFU / mL.

[0123] Preparation of AKK ONE live bacteria resuspension: The AKK ONE live bacteria suspension obtained above was pasteurized (80° C., 30 min, water bath) to obtain a dead bacteria suspension.

[0124] Culture of Mycoplasma pneumoniae: Mycoplasma pneumoniae (ATCC 15531) was revived in a sterile environment, cultured in a 37°C, 5% CO2 incubator, and subcultured every 6 days.

[0125] 1.1 Animal model construction and drug administration

[0126] 6-week-old SPF BALB / c mice, weighing 20-22 g, were purchased from Hunan Slake Jingda Experimental Animal Co., Ltd., with the catalog number: hnslkjd004; ambroterol oral solution, manufactured by Beijing Hanmei Pharmaceutical Co., Ltd., with the approval number: National Medicine Standard H20040317.

[0127] Thirty-five BALB / c mice were randomly divided into a control group, a model group, an ambroxol oral solution group, an AKK ONE dead bacteria resuspension group, and an AKK ONE live bacteria resuspension group, with 7 mice in each group. Except for the control group, mice in each group were anesthetized with ether and 20 μL of 1×10 6 Mycoplasma pneumoniae, once a day, for 5 days. After the model was confirmed to be successful, each group was given medication for 6 consecutive days, once a day, among which the ambroxol oral solution group (1.5 mL / Kg), the AKK ONE dead bacteria resuspension group (1×10 9 TFU / Kg), AKK ONE live bacteria resuspension (1×10 9 CFU / Kg); the control group and the model group were given the same volume of normal saline (1.5mL / Kg).

[0128] 1.2 Detection indicators

[0129] 1.2.1 Cough induction experiment

[0130] 40 minutes after the last administration, a beaker was placed upside down in the clean bench. A cotton ball containing 0.5 mL of ammonia was attached to the top of the beaker. The mouse was placed in the beaker and the timer was started. The number of coughs of the mouse within 5 minutes was observed and recorded. After the coughing experiment was completed, the mouse was killed and its lung tissue was taken for subsequent experiments.

[0131] 1.2.2 Enzyme-linked immunosorbent assay to determine the expression of related factors in serum

[0132] IL-6 ELISA test kit, purchased from Shanghai Saipeison Biotechnology Co., Ltd., catalog number: SPS-16497;

[0133] IL-8 ELISA test kit, purchased from Hefei Lyle Biotechnology Co., Ltd., catalog number: LE-B1155;

[0134] IL-10 ELISA detection kit, purchased from Shanghai Zeye Biotechnology Co., Ltd., item number: ZY72237H.

[0135] Enzyme-linked immunosorbent assay was used to determine IL-6, IL-8 and IL-10 in mouse serum. The experiment of the present invention was carried out strictly according to the steps in the operating instructions of the ELISA kit.

[0136] 1.2.3 mRNA expression levels of TRPA1, SP, and CGRP

[0137] Lung tissues of mice in each group were taken, and total RNA of lung tissues of mice in each group was extracted according to RNA extraction kit (Acori Biotechnology, AG21017), and RNA concentration of each group was detected by microplate reader. The expression levels of TRPA1, SP, and CGRP mRNA were detected using UltraSYBR One Step RT-qPCR Kit (purchased from Shanghai Bangjing Industrial Co., Ltd.; catalog number BJ-S963777) according to the kit instructions.

[0138] Taking β-actin as the internal reference, the relative mRNA expression of each target gene was calculated according to the formula 2-ΔΔCT. The primer gene sequences of TRPA1, SP, and CGRP are shown in Table 2 below:

[0139] Table 2

[0140]

[0141]

[0142] 1.3 Test results and analysis

[0143] 1.3.1 Cough induction experiment

[0144] Results Figure 1 Compared with the control group, the number of coughs within 5 minutes in the model group mice increased significantly (p<0.01). After drug intervention, the number of coughs within 5 minutes in the mice decreased significantly. The effects of the ambroxol oral solution group and the AKK ONE dead bacteria resuspension group were comparable, and there was no significant difference between the two groups and the control group. The effect of the AKK ONE live bacteria resuspension group was not as good as that of the AKK ONE dead bacteria resuspension group (p<0.05).

[0145] 1.3.2 Enzyme-linked immunosorbent assay to determine the expression of related factors in serum

[0146] The test results are shown in Table 1.

[0147] Table 1

[0148] Group IL-6 (pg / mL) IL-8 (pg / mL) IL-10 (pg / mL) Control group 20.32±3.53 15.81±1.82 384.56±91.42 Model Group 87.81±7.83 59.17±6.37 410.22±101.33 Positive drug control group 37.43±4.81 24.69±3.76 779.42±125.86 AKK ONE dead bacteria resuspension group 36.15±3.47 27.45±4.01 836.57±130.44 AKK ONE live bacteria resuspension group 49.94±5.95 37.84±2.69 695.46±109.85

[0149] From the data in Table 1, we can see that compared with the control group, the expression levels of IL-6 and IL-8 in the serum of the model group mice were significantly increased (p<0.01), and drug intervention could significantly downregulate the expression levels of IL-6 and IL-8. Among them, there was no significant difference between the positive drug control group and the AKKONE dead bacteria resuspension group, and the downregulation degree of the two groups was the largest and the effect was the best. The downregulation effect of the AKK ONE live bacteria resuspension group on proinflammatory factors was poorer than that of the AKK ONE dead bacteria resuspension group.

[0150] The positive drug control group, AKK ONE dead bacteria resuspension group and AKK ONE live bacteria resuspension group could all upregulate the expression of the anti-inflammatory factor IL-10, and the upregulation effect of the AKK ONE dead bacteria resuspension group was better than that of the positive drug group (p<0.05), but the AKK ONE live bacteria resuspension group was worse than that of the positive drug group (p<0.05).

[0151] 1.3.3 mRNA expression levels of TRPA1, SP, and CGRP

[0152] The results of the detection of mRNA expression levels of TRPA1, SP, and CGRP are shown in Table 3:

[0153] Table 3

[0154]

[0155]

[0156] From the data in Table 3, we can see that compared with the control group, the mRNA expression levels of TRPA1, SP, and CGRP in the lung tissue of the model group mice were significantly upregulated (p<0.01). After drug intervention, the mRNA expression levels of TRPA1, SP, and CGRP in the lung tissue of mice were downregulated to varying degrees, among which the AKK ONE dead bacteria resuspension group had the largest degree of downregulation, which was significantly better than the positive drug group (p<0.01), and had no significant difference with the control group, and was even better than the control group; the expression level of the AKK ONE live bacteria resuspension group was significantly different from that of the positive drug group (p<0.05), and the effect was slightly lower than that of the positive drug group.

Claims

1. Akkermansia muciniphila Akkermansia muciniphila ) Use of AKK ONE in the preparation of postbiotics for preventing and / or treating mycoplasma pneumonia, characterized in that: The deposit number of the AKK ONE is CGMCC No. 20954; the postbiotics are selected from the live bacteria suspension and / or dead bacteria suspension of AKK ONE.

2. The use according to claim 1, characterized in that: The preparation method of the postbiotics includes culturing AKKONE.

3. The use according to claim 2, characterized in that the culture medium used for culturing AKK ONE is BHI culture medium.

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