The use of Akkermansia muciniphila in improving cognitive function

Abstract

The present invention belongs to the field of microbial technology, and specifically relates to the application of Akkermansia muciniphila in improving cognitive function. The present invention provides the application of Akkermansia muciniphila in improving cognitive function, and the Akkermansia muciniphila is deposited in CGMCC with a deposit number of CGMCC No.20955. The Akkermansia muciniphila can significantly improve cognitive function, increase the content of anti-inflammatory factors IL-4 and IL-10, significantly downregulate the expression of inflammatory factors and apoptosis factors, significantly upregulate the expression of Aβ degrading enzyme IDE, protect the normal morphology of the colon, and is non-toxic to the liver.

Description

Technical Field

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

[0002] Neurodegenerative diseases are a type of disease in which the motor or psychological functions of the central or peripheral nervous system are gradually impaired. Their pathological characteristics are abnormal aggregation and deposition of proteins in the brain and spinal cord. Neurodegenerative diseases mainly include Parkinson's disease (PD), Alzheimer's disease (AD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). These diseases have similar histopathological characteristics and common cellular and molecular mechanisms. Currently known neurodegenerative diseases are divided into two types according to the symptoms of the patient at the time of onset: one type will damage the patient's memory and cognitive ability, and eventually cause dementia in the patient; the second type affects the patient's movement and balance ability.

[0003] Huntington's disease, the main disease population is middle-aged people. Huntington's disease can damage the patient's memory and cognitive ability, as well as affect the patient's motor ability. The main manifestations are chorea-like movements and neurological dysfunction, and patients usually die within 10 to 20 years after the onset of the disease. Frontotemporal dementia, generally occurs in the early elderly, mainly the frontal bone and temporal lobe area of ​​the patient's brain degeneration and atrophy, thereby damaging the patient's language ability and motor ability. Frontotemporal degeneration can lead to dementia in patients three to four years after the onset of the disease. Amyotrophic lateral sclerosis is a neurodegenerative disease that occurs frequently in adults. Some cases of the disease may be related to genetic and familial gene defects. The main feature of ALS is the death of motor neurons. As the disease progresses, symptoms such as respiratory failure, muscle atrophy, and dysphagia will appear. Generally, patients will die of respiratory failure 3 to 5 years after the onset of the disease.

[0004] Alzheimer's disease (AD) is the most common type of dementia in the elderly. Its clinical manifestations are deterioration of cognitive function and memory ability. It accounts for 50%-75% of all dementia and is also the neurodegenerative disease with the highest incidence (Thies, et al. 2012 Alzheimer's disease facts and figures. [J]. Alzheimers & Dementia the Journal of the Alzheimers Association, 2012, 8 (2): 131-168.). The brains of AD patients show global brain atrophy, with dense senile plaques (SP) formed by the accumulation of amyloid beta peptide (Aβ) aggregates, neurofibrillary tangles (NFTs) caused by hyperphosphorylation of microtubule-associated protein tau, and massive neuronal loss (Jakob Roetne, et al. Alzheimer's disease: From pathology to therapeutic approaches. [J]. Angew Chem Int Ed Engl, 2010, 40(25): 3030-3059.).

[0005] There are many theories about the pathogenesis of Alzheimer's disease, and the most widely accepted one is the "amyloid cascade hypothesis". The amyloid hypothesis was first proposed by Hardy et al. in 1992. One of the characteristic pathological changes of AD is the formation of senile plaques, and the core component of senile plaques is β-amyloid protein. When neurotoxic Aβ is abnormally deposited in the brain, senile plaques will form. Abnormal accumulation and aggregation of Aβ can initiate a pathological cascade reaction, inducing abnormal phosphorylation of Tau and subsequent formation of neurofibrillary tangles, inducing neuroinflammation and oxidative stress in the brain, and then leading to widespread neuronal loss (Hardy JA, et al. Alzheimer's disease: The amyloid cascade hypothesis. [J]. Science, 1992, 256(5054): 184-185.; Hardy, et al. Alzheimer's disease: The amyloid cascade hypothesis: An update and reappraisal. [J]. Journal of Alzheimers Disease, 2006, 9(S3): 151-153.).

[0006] Aging is the main risk factor for neurodegenerative diseases, which seriously endanger the health of the elderly. Therefore, it is of great significance to find safe and effective treatments.

[0007] In recent years, with the in-depth study of intestinal flora, the theory of microbial-gut-brain axis has been proposed. Intestinal microorganisms regulate gastrointestinal function and can also affect the function of the central nervous system through the neuro-immune-endocrine system. Studies have found that the intestinal microbiome changes in various neurodegenerative diseases, indicating that the occurrence and development of neurodegenerative diseases has a significant impact on the intestinal microbiome (LOPEZ-OTIN C, et al. The hallmarks of aging. [J]. Cell, 2013, 153 (6): 1194.121). Therefore, the use of microorganisms to improve and / or treat neurodegenerative diseases has become a new option.

[0008] U.S. patent application US17780436 discloses a composition and method for treating or preventing the progression of a neurodegenerative disease, the composition comprising a bacterial composition having an effective amount of live non-pathogenic microorganisms, live non-pathogenic bacteria, wherein at least one of the bacteria is a predatory bacterium, such as Bdellovibrio; the disclosed bacterial composition can be used to treat or prevent the progression of neurodegenerative diseases such as ALS, Alzheimer's disease, Huntington's disease and Parkinson's disease.

[0009] Chinese patent application 201880039628.9 discloses a composition comprising a bacterial strain of the genus Megasphaera, which is used to treat or prevent neurodegenerative disorders. The composition is used to treat or prevent early-onset neurodegenerative diseases, and can also be used to prevent or delay the onset or progression of neurodegenerative disorders.

[0010] In the field of neurodegenerative diseases, especially Alzheimer's disease, there is still a need for more microbial methods and drugs for treating and / or preventing neurodegenerative diseases. Summary of the invention

[0011] In order to overcome the defects of the prior art, the present invention hereby proposes a use of Akkermansia muciniphila PROBIO (hereinafter referred to as AKK PROBIO) in improving cognitive function.

[0012] The inventor creatively isolated a strain of Akkermansia muciniphila from a healthy human body, which has been deposited in CGMCC with the deposit number CGMCC No.20955. The inventor also identified and characterized its morphological taxonomy and biochemical genetic functions, and explored its applications in different fields (other patent applications have been submitted on the same day).

[0013] The inventors were surprised to find that the application of AKK PROBIO in multiple fields such as anti-tumor, cognitive disease, metabolic disease, inflammatory disease, bone and joint disease, cardiovascular disease, etc. has achieved unexpected positive effects. However, considering the practice of Chinese patent law, we now apply for protection for each indication that does not have a single nature.

[0014] This application is to protect the application of AKK PROBIO in improving cognitive function.

[0015] Based on the practice of Chinese patent law, in order to avoid the same inventions and creations submitted on the same day, the applicant only recorded the relevant experimental data such as microbial morphological taxonomic identification and biochemical genetic performance characterization in the microbial patent application submitted on the same day. This application does not record the relevant information of the microorganism, nor does it protect the microorganism itself. In order to facilitate the review of the present invention and quickly understand the inventor's previous work, the relevant information of AKK PROBIO is now briefly disclosed in the invention content of this application.

[0016] 1. Basic information of AKK PROBIO

[0017] 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.

[0018] 1. Akkermansia muciniphila CGMCC No.20955 was screened and isolated by the inventors;

[0019] 2. The deposit information of Akkermansia muciniphila CGMCC No.20955;

[0020] 3. 16s rRNA of Akkermansia muciniphila CGMCC No.20955;

[0021] 4. Physiological and biochemical properties of Akkermansia muciniphila CGMCC No.20955:

[0022] (1) Hydrophobicity: increases with time, reaching more than 30% at 60 min;

[0023] (2) Self-aggregation: It increases with time and stabilizes at around 52% after 20 hours.

[0024] (3) Gastrointestinal fluid tolerance:

[0025] The survival rate in gastric juice showed a decreasing trend with the extension of time, and the survival rate was about 85% at 240 minutes;

[0026] The survival rate in intestinal fluid showed an overall downward trend, and at 240 min the survival rate was above 80%;

[0027] (4) Biofilm formation ability: weak;

[0028] (5) Acute toxicity test:

[0029] The results of bacterial reverse mutation test showed that the strain had no mutagenicity;

[0030] Mice were acutely gavaged with high, medium, and low doses of Akkermansia muciniphila CGMCC No.20955, and no mortality occurred;

[0031] There was no significant difference in the daily body weight changes between the test group mice and the control saline group;

[0032] 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;

[0033] 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;

[0034] There were no significant differences in the blood parameters between the acute gavage group and the control saline group;

[0035] In the liver, the triglyceride levels in the test mice were significantly lower than those in the control saline group mice;

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

[0037] There was no significant difference in cholesterol levels in serum and liver between the test mice and the control mice;

[0038] There was no significant difference in the bile acid levels in serum and liver between the test mice and the control mice;

[0039] There was no significant difference in glucose levels in serum and liver between the test mice and the control mice;

[0040] There was no significant difference in the total protein levels in serum and liver between the test mice and the control mice;

[0041] There was no significant difference in the activity of alanine aminotransferase in serum and liver between the test mice and the control mice.

[0042] There was no significant difference in the activity of aspartate aminotransferase in serum and liver between the test mice and the control mice;

[0043] There was no significant difference in serum creatinine content between the test mice and the control mice;

[0044] There was no significant difference in the urea nitrogen concentration in the serum between the test mice and the control mice;

[0045] 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;

[0046] There was no significant pathological damage in the liver and kidneys of the test mice and the control group mice.

[0047] (6) Subchronic toxicity test:

[0048] Mice were gavaged with high, medium, and low doses of AKK PROBIO for 90 consecutive days without mortality.

[0049] There was no significant difference in the weekly body weight change between the test group mice and the control saline group;

[0050] 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;

[0051] During the continuous gavage period, the changes in blood glucose in the mice given a medium dose of oral administration were significantly different from those in the control saline group, but there were no significant differences between the other groups and the control group.

[0052] There were no significant differences in the blood parameters between the mice in the gavage group and the control saline group;

[0053] There was no significant difference in triglyceride levels in the liver and serum between the test mice and the control mice;

[0054] There was no significant difference in cholesterol levels in the liver and serum between the test mice and the control mice;

[0055] The bile acid levels in the liver of the mice given the medium dose and 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.

[0056] There was no significant difference in glucose levels in the liver and serum between the test mice and the control mice;

[0057] There was no significant difference in the total protein content in the liver and serum between the test mice and the control mice;

[0058] There was no significant difference in the activity of alanine aminotransferase in serum and liver between the test mice and the control mice.

[0059] There was no significant difference in the activity of aspartate aminotransferase in serum and liver between the test mice and the control mice;

[0060] There was no significant difference in serum creatinine content between the test mice and the control mice;

[0061] There was no significant difference in the urea nitrogen concentration in the serum between the test mice and the control mice;

[0062] 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;

[0063] There was no significant pathological damage in the liver and kidneys of the test mice and the control mice;

[0064] There was no significant difference in blood sugar regulation ability between the test mice and the control group mice.

[0065] The technical purpose of the present invention is to provide a microorganism that can improve cognitive function.

[0066] (7) Drug sensitivity analysis:

[0067] AKK PROBIO was found to be sensitive to ampicillin, ceftriaxone, ceftriaxone, meropenem, tetracycline, moxifloxacin and chloramphenicol.

[0068] Gut microbiota refers to the microorganisms that grow in the human digestive tract. It is numerous and complex in variety, and is composed of bacteria, fungi, viruses, and protozoa. Since bacteria are the most widely studied, the intestinal microbiota is also commonly referred to as the intestinal flora (Backhed F, et al. Host-bacterial mutualism in the human intestine. [J]. Science, 2005, 307 (5717): 1915-1920.). The intestinal flora not only assists the host in digesting food and providing nutrients, but also has a profound impact on the host's health. Studies have shown that the structure and composition of different intestinal microorganisms affect the host's nutrient processing, energy balance, immune function, gastrointestinal development, and other important physiological activities. The intestinal flora plays an important role in our growth, development, metabolism, and other aspects (Wen L, et al. Innate immunity and intestinal microbiota in the development of Type 1 diabetes. [J]. Nature, 2008, 455 (7216): 1109-1113.).

[0069] Intestinal flora plays an important role in regulating host metabolism and low-grade inflammation. Therefore, the goal of improving human health can be achieved by regulating the structure of intestinal flora. Probiotics developed based on intestinal native bacteria are expected to become a new way to treat many diseases. Probiotics refer to the general term for beneficial microorganisms that colonize and survive in the human intestine after the host consumes enough and can produce definite health effects. Different types of probiotics play different functions. Probiotics are roughly divided into traditional lactic acid probiotics, non-lactic acid probiotics and next-generation probiotics. Traditional probiotics mainly play a preventive and health-care role, while the research direction of next-generation probiotics focuses on the treatment and cure of diseases (Botina SG, et al. Classification of domestic probiotic cultures of Lactobacillus genus. [J]. Zh Mikrobiol Epidemiol Immunobiol, 2010 (5): 3-7.). Among them, the next-generation probiotics mainly include Akkermansia, Bacteroides fragilis, Clostridium perfringens, etc. As a potential probiotic, Akkermansia muciniphila is the main research object of the next generation of probiotics and has broad prospects for medical applications.

[0070] Akkermansia muciniphila is an oval, flagella-free, non-spore-forming Gram-negative anaerobic bacterium with a single cell long axis of 0.6 μm-1.0 μm. It can grow alone or in pairs, and can also grow in groups in a culture medium containing mucin. The colonies present a white biofilm, and the colonies are connected by filamentous structures. The size of a single colony is about 0.7 mm (Derrien M. et al. Akkermansia muciniphila gen. nov., sp. nov., a human intestinal mucin-degrading bacterium. [J]. INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2004, 54 (5): 1469-1476.). Akkermansia muciniphila is a typical intestinal anaerobic bacterium. Its isolation and purification are extremely difficult, and the strains successfully isolated are very limited. Currently, there are only two species of Akkermansia, Akkermansia muciniphila isolated from human feces and Akkermansia glycaniphila isolated from reticulated python feces (Ouwerkerk JP, et al. Akkermansia glycaniphila sp.nov., an anaerobic mucin-degrading bacterium isolated from reticulated python faeces [J]. International Journal of Systematic and Evolutionary Microbiology, 2016, 66 (11): 4614-4620.). However, Akkermansia muciniphila is a potential probiotic, and obtaining its pure culture strain is necessary to clarify the function of the bacterium.

[0071] In order to achieve the above technical objectives, the present invention hereby proposes the following technical solutions:

[0072] In one aspect, the present invention provides use of Akkermansia muciniphila in improving cognitive function. The Akkermansia muciniphila is deposited in CGMCC with a deposit number of CGMCC No.20955.

[0073] In one embodiment, the Akkermansia muciniphila improves cognitive function via at least one of the following mechanisms:

[0074] (1) Promote the expression of insulin-degrading enzyme IDE;

[0075] (2) Increase the levels of anti-inflammatory factors IL-4 and IL-10;

[0076] (3) downregulate the expression of proinflammatory factors IL-1β, IL-18, CCL2, NLRP3 and chemokine CXCL1;

[0077] (4) Down-regulate the expression of Aβ protein.

[0078] In another aspect, Akkermansia muciniphila is used in the preparation of a drug or a pharmaceutical raw material for preventing, treating and / or assisting in the treatment of Alzheimer's disease. The Akkermansia muciniphila is deposited in CGMCC with a deposit number of CGMCC No.20955.

[0079] In one embodiment, the drug or drug substance improves cognitive function through at least one of the following mechanisms:

[0080] 1) Promote the expression of insulin-degrading enzyme IDE;

[0081] 2) Increase the levels of anti-inflammatory factors IL-4 and IL-10;

[0082] 3) down-regulate the expression of pro-inflammatory factors IL-1β, IL-18, CCL2, NLRP3 and chemokine CXCL1;

[0083] 4) Down-regulate the expression of Aβ protein.

[0084] In a preferred embodiment, the neurodegenerative disease may be Alzheimer's disease.

[0085] In one embodiment, the product comprises living or dead forms of Akkermansia muciniphila, isolates and / or secretions thereof.

[0086] In one embodiment, the living form of Akkermansia muciniphila can be any biologically active Akkermansia muciniphila, such as live bacteria in fermentation broth, live bacteria in solid culture medium, live bacteria in fermentation broth precipitate, live bacteria in fermentation broth supernatant, live bacteria naturally existing in nature, live bacteria existing in the intestines of humans or animals, etc.

[0087] In one embodiment, the dead form of Akkermansia muciniphila can be any Akkermansia muciniphila without biological activity, such as dead bacteria in fermentation broth, dead bacteria in solid culture medium, dead bacteria in fermentation broth precipitate, dead bacteria in fermentation broth supernatant, dead bacteria naturally existing in nature, dead bacteria existing in the intestines of humans or animals, etc.

[0088] In one embodiment, the probiotic preparation comprises an effective amount of a living form or a dead form of Akkermansia muciniphila, and an isolate and / or secretion thereof, wherein the deposit number of Akkermansia muciniphila is CGMCC No.20955.

[0089] In one embodiment, the probiotic preparation further comprises at least one of a food-acceptable excipient, a food-acceptable compound, a food-acceptable protein and / or a food-acceptable polypeptide.

[0090] In a preferred embodiment, the food-acceptable excipient is selected from at least one of erythritol, D-mannitol, fumaric acid, glycerol, pectin, potassium alginate, sodium alginate, talc, sodium pyrophosphate, polydextrose, carrageenan, sodium ascorbate, ascorbyl palmitate, L-malic acid, L(+)-tartaric acid, maltitol, gelatin, xylitol, citric acid, potassium citrate, sodium citrate, citric acid fatty acid glyceride, agar, lactic acid, sodium lactate, sorbic acid and its potassium salt, sorbitol, acid red, calcium carbonate, sodium carbonate, sodium bicarbonate, beet red, vitamin C, vitamin E, oxidized starch, ethanol, sodium acetate, stearic acid, calcium stearate, magnesium stearate, α-cyclodextrin, γ-cyclodextrin and dextrin.

[0091] In a preferred embodiment, the food acceptable compound can be in the form of a compound of trace elements beneficial to the human body and / or their derivatives, such as iron sulfate, succinic acid, zinc sulfate, zinc gluconate, etc.; compounds beneficial to the human body, such as β-carotene, polyphenol antioxidants, dietary fiber, etc.

[0092] In a preferred embodiment, the food acceptable protein and / or polypeptide includes soy protein isolate, meat protein, cereal protein, milk protein, plant polypeptide, animal polypeptide and the like.

[0093] In one embodiment, the dosage form of the probiotic preparation includes a liquid preparation, a solid preparation, a semi-solid preparation or a gaseous preparation.

[0094] In a preferred embodiment, the liquid preparation can be a liquid seasoning, a liquid dairy product, a liquid beverage, or the like.

[0095] Among them, liquid dairy products may be pasteurized milk, high-temperature sterilized milk, formulated milk, sterilized milk, fermented milk, condensed milk, cream, light cream, anhydrous butter, specialty dairy products, concentrated milk, etc.;

[0096] Liquid beverages may be fruit juice, vegetable juice, pulp, compound fruit and vegetable juice, concentrated fruit and vegetable juice, milk-containing beverages, plant protein beverages, compound protein beverages, coffee beverages, plant beverages, flavored beverages, sports drinks, nutrient beverages, energy drinks, electrolyte beverages, etc.

[0097] In a preferred embodiment, the solid preparation can be solid condiments, solid dairy products, solid beverages, biscuits, candy products, frozen drinks, etc.

[0098] Among them, solid dairy products can be whole milk powder, skimmed milk powder, partially skimmed milk powder, reconstituted milk powder, whey powder, cheese, processed cheese, etc.;

[0099] The solid beverage may be a protein solid beverage, a fruit and vegetable solid beverage, a tea solid beverage, or a coffee solid beverage;

[0100] In a preferred embodiment, the biscuits include crisp biscuits, tough biscuits, fermented biscuits, compressed biscuits, cookies, sandwich (filled) biscuits, wafer biscuits, egg round biscuits, egg rolls, pancakes, decorated biscuits, and bubble biscuits.

[0101] In a preferred embodiment, the frozen beverage includes ice cream, slush, popsicles, edible ice, and sweet ice.

[0102] In one embodiment, the semi-solid formulation includes beverage concentrates, ice cream, and the like.

[0103] In one embodiment, each unit of the probiotic preparation contains at least about 10 8 CFU / g or CFU / mL.

[0104] Among them, each unit can refer to each tablet, each dose, each pill, each pill drop, each capsule and the like. Those skilled in the art should know that the unit depends on the dosage form / carrier form of the preparation and does not constitute a limitation on the scope of protection of the present invention.

[0105] In a preferred embodiment, each unit of the probiotic preparation contains about 10 8 -10 12 CFU / g or CFU / mL.

[0106] In a preferred embodiment, each unit of the probiotic preparation contains about 10 9 -10 12 CFU / g or CFU / mL.

[0107] In a preferred embodiment, each unit of the probiotic preparation contains about 10 9 -10 11 CFU / g or CFU / mL.

[0108] In a preferred embodiment, each unit of the probiotic preparation may contain about 10 9 -10 10 CFU / g or CFU / mL.

[0109] In a preferred embodiment, each unit of the probiotic preparation may contain about 1.0×10 9 , 1.1×10 9 , 1.2×10 9 , 1.3×10 9 , 1.4×10 9 , 1.5×10 9 , 1.6×10 9 , 1.7×10 9 , 1.8×10 9 , 1.9×10 9 , 2.0×10 9 , 2.1×10 9 , 2.2×10 9 , 2.3×10 9 , 2.4×10 9 , 2.5×10 9 , 2.6×10 9 , 2.7×10 9 , 2.8×10 9 , 2.9×10 9 , 3.0×10 9 , 3.1×10 9 , 3.2×10 9 , 3.3×10 9 , 3.4×10 9 , 3.5×10 9 , 3.6×10 9 , 3.7×10 9 , 3.8×10 9 , 3.9×109 、4.0×10 9 、4.1×10 9 、4.2×10 9 、4.3×10 9 、4.4×10 9 、4.5×10 9 、4.6×10 9 、4.7×10 9 、4.8×10 9 、4.9×10 9 、5.0×10 9 、5.1×10 9 、5.2×10 9 、5.3×10 9 、5.4×10 9 、5.5×10 9 、5.6×10 9 、5.7×10 9 、5.8×10 9 、5.9×10 9 、6.0×10 9 、6.1×10 9 、6.2×10 9 、6.3×10 9 、6.4×10 9 、6.5×10 9 、6.6×10 9 、6.7×10 9 、6.8×10 9 、6.9×10 9 、7.0×10 9 、7.1×10 9 、7.2×10 9 、7.3×10 9 、7.4×10 9 、7.5×10 9 、7.6×10 9 、7.7×10 9 、7.8×10 9 、7.9×10 9 、8.0×10 9 、8.1×10 9 、8.2×10 9 、8.3×10 9 、8.4×10 9 、8.5×10 9 、8.6×10 9 、8.7×10 9 、8.8×10 9 、8.9×109 9.0×10 9 , 9.1×10 9 , 9.2×10 9 , 9.3×10 9 , 9.4×10 9 , 9.5×10 9 , 9.6×10 9 , 9.7×10 9 , 9.8×10 9 9.9×10 9 , 1.0×10 10 CFU / g or CFU / mL.

[0110] In another aspect, the present invention provides a drug for preventing and / or treating neurodegenerative diseases, comprising an effective amount of a living form or a dead form of Akkermansia muciniphila, and an isolate and / or secretion thereof, wherein the deposit number of Akkermansia muciniphila is CGMCC No.20955.

[0111] In one embodiment, the neurodegenerative disease may be Alzheimer's disease.

[0112] In one embodiment, the living form of Akkermansia muciniphila can be any biologically active Akkermansia muciniphila, for example: live bacteria in fermentation broth, live bacteria in solid culture medium, live bacteria in fermentation broth precipitate, live bacteria in fermentation broth supernatant, live bacteria naturally existing in nature, live bacteria existing in the intestines of humans or animals, live bacteria in freeze-dried powder, etc.

[0113] In one embodiment, the dead form of Akkermansia muciniphila can be any Akkermansia muciniphila without biological activity, for example: dead bacteria in fermentation broth, dead bacteria in solid culture medium, dead bacteria in fermentation broth precipitate, dead bacteria in fermentation broth supernatant, dead bacteria naturally existing in nature, dead bacteria existing in the intestines of humans or animals, dead bacteria in freeze-dried powder, etc.

[0114] In a preferred embodiment, the pharmaceutically acceptable excipient is selected from at least one of a solvent, a diluent, a disintegrant, a precipitation inhibitor, a surfactant, a glidant, a binder, a lubricant, a dispersant, a suspending agent, an isotonic agent, a thickener, an emulsifier, a preservative, a stabilizer, a hydrating agent, an emulsification accelerator, a buffer, an absorbent, a colorant, a flavoring agent, a sweetener, an ion exchanger, a release agent, a coating agent, a flavoring agent and an antioxidant.

[0115] In a preferred embodiment, the dosage form of the drug is selected from at least one of powders, tablets, granules, capsules, solutions, emulsions, suspensions, injections, sprays, powder sprays, aerosols, suppositories, drops and pills.

[0116] In one embodiment, each unit of the drug contains at least about 10 8 CFU / g or CFU / mL.

[0117] Among them, each unit can refer to each tablet, each dose, each pill, each pill, each capsule and the like. Those skilled in the art should know that the unit depends on the dosage form / carrier form of the drug and does not constitute a limitation on the scope of protection of the present invention.

[0118] In a preferred embodiment, each unit of the drug may contain about 10 8 -10 12 CFU / g or CFU / mL.

[0119] In a preferred embodiment, each unit of the drug may contain about 10 9 -10 12 CFU / g or CFU / mL.

[0120] In a preferred embodiment, each unit of the drug may contain about 10 9 -10 11 CFU / g or CFU / mL.

[0121] In a preferred embodiment, each unit of the drug may contain about 10 9 -10 10 CFU / g or CFU / mL.

[0122] In a preferred embodiment, each unit of the probiotic preparation may contain about 1.0×10 9 , 1.1×10 9 , 1.2×10 9 , 1.3×10 9 , 1.4×10 9 , 1.5×10 9 , 1.6×10 9 , 1.7×10 9 , 1.8×109 、1.9×10 9 、2.0×10 9 、2.1×10 9 、2.2×10 9 、2.3×10 9 、2.4×10 9 、2.5×10 9 、2.6×10 9 、2.7×10 9 、2.8×10 9 、2.9×10 9 、3.0×10 9 、3.1×10 9 、3.2×10 9 、3.3×10 9 、3.4×10 9 、3.5×10 9 、3.6×10 9 、3.7×10 9 、3.8×10 9 、3.9×10 9 、4.0×10 9 、4.1×10 9 、4.2×10 9 、4.3×10 9 、4.4×10 9 、4.5×10 9 、4.6×10 9 、4.7×10 9 、4.8×10 9 、4.9×10 9 、5.0×10 9 、5.1×10 9 、5.2×10 9 、5.3×10 9 、5.4×10 9 、5.5×10 9 、5.6×10 9 、5.7×10 9 、5.8×10 9 、5.9×10 9 、6.0×10 9 、6.1×10 9 、6.2×10 9 、6.3×10 9 、6.4×10 9 、6.5×10 9 、6.6×10 9 、6.7×10 9 、6.8×109 , 6.9×10 9 , 7.0×10 9 , 7.1×10 9 , 7.2×10 9 , 7.3×10 9 , 7.4×10 9 , 7.5×10 9 , 7.6×10 9 , 7.7×10 9 , 7.8×10 9 ,7.9×10 9 , 8.0×10 9 , 8.1×10 9 , 8.2×10 9 , 8.3×10 9 , 8.4×10 9 , 8.5×10 9 , 8.6×10 9 , 8.7×10 9 , 8.8×10 9 , 8.9×10 9 9.0×10 9 , 9.1×10 9 , 9.2×10 9 , 9.3×10 9 , 9.4×10 9 , 9.5×10 9 , 9.6×10 9 , 9.7×10 9 , 9.8×10 9 9.9×10 9 , 1.0×10 10 CFU / g or CFU / mL.

[0123] In yet another aspect, the present invention provides a method for preventing, treating and / or assisting in the treatment of a subject suffering from a neurodegenerative disease, wherein an effective amount of a living form or a dead form of Akkermansia muciniphila, an isolate and / or secretion thereof, the probiotic preparation or the medicament is administered to the subject;

[0124] The deposit number of the Akkermansia muciniphila is CGMCC No.20955.

[0125] In one embodiment, the neurodegenerative disease may be Alzheimer's disease.

[0126] Compared with the prior art, the Akkermansia muciniphila provided by the present invention has at least the following beneficial effects:

[0127] (1) Significantly improved cognitive function. In the water maze test, the latency was shortened by at least 10 seconds compared with the model group.

[0128] (2) The levels of anti-inflammatory factors IL-4 and IL-10 were significantly increased, and the levels of pro-inflammatory factors CXCL1, CCL2, and Aβ were downregulated. Compared with the model group, IL-4 increased by about 2.5 times, IL-10 increased by about 1.4 times, CXCL1 level was downregulated by about 51%, CCL2 level was downregulated by about 25%, and Aβ level was downregulated by about 50%;

[0129] (3) The expression of pro-inflammatory factors (IL-1β, IL-18, NLRP3) and apoptosis factor Caspase-1 was significantly downregulated. Compared with the model group, the level of IL-1β was downregulated by about 46%, the level of IL-18 was downregulated by 44%, the level of NLRP3 was downregulated by about 40%, and the level of Caspase-1 was downregulated by about 47%. The expression of Aβ-degrading enzyme IDE was significantly upregulated, which was 2 times higher than that of the model group.

[0130] (4) Significantly down-regulated the expression level of Aβ protein in hippocampal tissue;

[0131] (5) It can protect the normal morphology of the colon and is non-toxic to the liver. BRIEF DESCRIPTION OF THE DRAWINGS

[0132] Figure 1 The latency statistics of the Morris water maze test are shown.

[0133] Figure 2 The trajectory diagram of the Morris water maze experiment. A is the trajectory diagram of the WT group mice from being placed in the pool until they find the platform, B is the trajectory diagram of the APP group mice from being placed in the pool until they find the platform, and C is the trajectory diagram of the APP+AKK PROBIO group mice from being placed in the pool until they find the platform.

[0134] Figure 3 The results of the determination of Aβ protein content in mice under different treatment groups. The letters a, b, and c represent significant differences, the same lowercase letters indicate no significant difference between the two groups, and different lowercase letters indicate significant difference between the two groups (p<0.05).

[0135] Figure 4 The figure shows the Western Blot results of Aβ protein in mice under different treatment groups.

[0136] Figure 5The liver sections of mice in different treatment groups. A is a liver section of mice in the WT group, B is a liver section of mice in the APP group, and C is a liver section of mice in the APP+AKK PROBIO group. Scale bar 100 μm.

[0137] Figure 6 The colon sections of mice in different treatment groups. A is a colon section of mice in the WT group, B is a colon section of mice in the APP group, and C is a colon section of mice in the APP+AKK PROBIO group. Scale bar 500 μm. DETAILED DESCRIPTION

[0138] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly used in the field to which the present invention belongs. For the purpose of interpreting this specification, the following definitions will apply, and where appropriate, terms used in the singular will also include the plural form, and vice versa.

[0139] Unless the context clearly dictates otherwise, the expressions "a", "an" and "an" as used herein include plural references. For example, reference to "a cell" includes a plurality of such cells and equivalents thereof known to those skilled in the art, and so forth.

[0140] As used herein, the term "about" refers to a range of ±20% of the value that follows. In some embodiments, the term "about" refers to a range of ±10% of the value that follows. In some embodiments, the term "about" refers to a range of ±5% of the value that follows.

[0141] The term "cognitive function" used in this article refers to the ability to recognize and know things, specifically including memory, calculation, comprehension thinking, judgment and reasoning, language expression, executive ability, time and space orientation, etc.

[0142] The term "Akkermansia muciniphila", "AKK", "AKK PROBIO" or "Akkermansia muciniphila" used herein refers to Akkermansia muciniphila with the deposit number of CGMCC No.20955.

[0143] The term "neurodegenerative disease" used in this article refers to a class of diseases in which damage to neurons in the brain and spinal cord gradually worsens over time, with a large loss of specific neurons as the main feature. It is a complex disease that progresses and eventually leads to death.

[0144] The term "viable form of Akkermansia muciniphila" as used herein refers to Akkermansia muciniphila having at least one of the following properties: culturability, transcribability, metabolic activity, and cell membrane integrity. Culturability refers to the ability of bacteria to form colonies on a culture medium; transcribability refers to the ability of bacterial cells to produce mRNA; metabolic activity refers to the ability of bacteria to exhibit life phenomena related to their metabolism, such as redox ability, respiration, active enzymes, and substrate uptake ability; and cell membrane integrity refers to the fact that the bacterial cell membrane is not damaged.

[0145] Otherwise, it is the "death form of Akkermansia muciniphila".

[0146] The term "isolate" as used herein refers to a microbial substance in which a specific microorganism is separated from a population or a mixed microbial population and exists in a certain medium, wherein the medium includes fermentation broth supernatant, fermentation broth precipitate, bacterial suspension, freeze-dried powder, etc.

[0147] The term "secretions" as used herein refers to products secreted by specific microbial individuals during growth or metabolism, including metabolites such as methanol, ethanol, propanol, isopropanol, n-butanol, succinic acid, tartaric acid, etc.; extracellular products such as proteins, fats, starch, cellulose, antibiotics, etc.

[0148] The term "food" as used herein refers to various finished products and raw materials for human consumption or drinking, as well as items that are traditionally both food and Chinese medicinal materials, but does not include items for therapeutic purposes.

[0149] The terms "health products", "dietary supplements" or "health foods" used in this article refer to a type of food that has the common properties of general foods, can regulate the functions of the human body, and is suitable for consumption by specific groups of people, but is not intended to treat diseases.

[0150] The term "food raw materials" used in this article refers to all raw materials used in food processing, including food staples, food ingredients, and food additives.

[0151] The term "drug raw material" or "pharmaceutical raw material" used in this article refers to the raw material drug containing the active ingredient of the drug and the biological and chemical substances that serve as its source.

[0152] The term "probiotic preparation" used in this article refers to a preparation prepared according to the principles of microecology using normal beneficial microorganisms or their promoting substances that are beneficial to the host. It has the function of maintaining or adjusting the balance of microecology, preventing and treating diseases and promoting the health of the host.

[0153] The term "subject" as used herein refers to a person who participates in a clinical trial and serves as a recipient of the trial drug, including patients and healthy subjects. Preferably, the subject is a patient.

[0154] The term "adjuvant therapy" as used herein refers to products that are not directly used to prevent or combat disease, but are used to reduce the side effects of treatment, improve patient tolerance, and enhance the effectiveness of treatment. Products used for adjuvant therapy generally refer to products that help promote the absorption of the main drug, prevent or reduce adverse reactions, and enhance body functions.

[0155] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with embodiment. If the specific conditions are not specified in the embodiment, it is carried out according to the conditions recommended by normal conditions or manufacturers. If the manufacturer is not specified in all reagents or instruments, they are all conventional products that can be purchased commercially. In order to better illustrate the present invention, numerous specific details are provided in the specific embodiments below. The specific embodiments described herein are only used to explain the present invention and are not intended to constitute any limitation of the present invention. In addition, in the following description, the description of known structures and technologies is omitted to avoid unnecessary confusion of the concept of the present invention. Such structures and technologies are also described in many publications.

[0156] Example 1 Cultivation of AKK PROBIO

[0157] Primary culture: Take AKK PROBIO frozen in a 2 mL glycerol tube, inoculate it at a 10% rate in 9 mL brain heart infusion broth (BHI) medium, and culture it anaerobically at 37°C for 24-48 hours to obtain a primary culture.

[0158] Secondary culture: Take the primary culture and inoculate it into a 90 mL Erlenmeyer flask at a 5% inoculation rate, and culture it anaerobically at 37°C for 24 hours to obtain the secondary culture.

[0159] Tertiary culture: Take the secondary culture and inoculate it into a 300 mL Erlenmeyer flask at a 5% inoculum size and culture it anaerobically at 37°C for 13 h.

[0160] Alternatively, add 20% glycerol to the tertiary culture and store frozen at -80°C.

[0161] Example 2 Application of AKK PROBIO in improving cognitive function

[0162] Preparation of main experimental reagents:

[0163] Preparation of AKK PROBIO bacterial suspension: Take 5 mL of AKK PROBIO frozen bacterial suspension for activation culture, centrifuge at 4000 r / min for 10 min, wash twice, and add 5 mL of physiological saline after washing to form a concentration of 1×10 10 CFU / mL of bacterial suspension.

[0164] The experiment of this embodiment was carried out in parallel for three tissue samples, and then the mean value was calculated. SPSS software (SPSS v.25 for Windows, IBM Software Group, Chicago, IL, USA) was used to average and analyze the data. The difference between the mean values ​​of each group was evaluated by one-way analysis of variance using Duncan's multiple range test. The difference of p < 0.05 was considered to be statistically significant.

[0165] 2.1 Animal Experiment Design

[0166] In this example, 20 12-month-old APP / PS1 double transgenic mice (Hangzhou Ziyuan Experimental Animal Technology Co., Ltd., MJ0324-058 batch) (Alzheimer's disease (AD) model mice) were randomly divided into 2 groups: APP group (model group) and APP+AKK PROBIO group (AKK PROBIO was administered to AD model mice), with 10 mice in each group; 10 wild-type C57 BL / 6J mice from the same litter were used as blanks, which were the WT group. All mice were gavaged and injected at 12 months of age for 30 days, as shown in Table 1. After the experiment, blood was collected from the orbital vein of the mice, and the mice were killed by cervical dislocation. Part of the brain, liver and colon tissues of the mice were taken in 4% paraformaldehyde, and the remaining tissues were stored in an ultra-low temperature refrigerator at -80°C.

[0167] Table 1. Groups and doses for oral administration

[0168]

[0169]

[0170] 2.2 Evaluation of AKK PROBIO in improving spatial memory ability

[0171] After the end of the experimental drug administration, a Morris water maze experiment was conducted for 4 days. The water maze consists of a circular pool (120 cm in diameter) and is equally divided into four quadrants. During the experiment, the mice were moved into the laboratory two hours in advance to familiarize themselves with the environment in advance. The water temperature was adjusted to about 21-22 ° C, and milk was poured into the pool to make it milky white. Days 1 to 3 were the training period. The mice were gently placed in the water from the first, second, third, and fourth quadrants facing the direction of the pool wall prompts. The swimming trajectory was recorded for 1 min. Mice that did not find the platform within the specified time needed to be guided to the platform. Each mouse was trained once in each quadrant every day for 3 consecutive days. Day 4 was the experimental period. Each mouse randomly selected a quadrant and placed it in the pool. The time required to find the platform within 60 seconds, that is, the escape latency, was recorded, and the swimming trajectory of each mouse was recorded with a video acquisition system; if the mouse failed to find the platform within 60 seconds after entering the water, the escape latency was recorded as 60 seconds.

[0172] The experimental results showed that after 4 consecutive days of Morris water maze test, the escape latency of each group of mice showed a decreasing trend ( Figure 1 ). Compared with the test results of the APP group on the last day, the escape latency of mice in the APP+AKK PROBIO group was shortened by about 22.5%, and the path to find the platform was also significantly shortened ( Figure 1 and Figure 2 ).

[0173] 2.3 Determination of the effect of AKK PROBIO on inflammatory markers

[0174] The hippocampal tissue of each group of mice in this embodiment was homogenized with PBS and the supernatant was aspirated for the experiment. The ELISA detection kit instructions detected the levels of IL-4 (interleukin-4, Nanjing Jiancheng Bioengineering Institute, item number: H005-1-2), IL-10 (interleukin-10, Nanjing Jiancheng Bioengineering Institute, item number: H009-1-2), CCL2 (chemokine ligand 2, Nanjing Jiancheng Bioengineering Institute, item number: H318-1) and CXCL1 (CXC chemokine ligand 1, Nanjing Jiancheng Bioengineering Institute, item number: H306-1) in the hippocampal tissue.

[0175] The results are shown in Table 2. The levels of CXCL1 and CCL2 in the hippocampus of the WT group were the lowest among the four groups, and the levels of IL-4 and IL-10 were the highest among the four groups. The levels of IL-4 and IL-10 in the APP group were the lowest among the four groups, and the levels of CXCL1 and CCL2 were the highest among the four groups. The levels of CXCL1 and CCL2 in the APP+AKK PROBIO group showed a downward trend compared with the APP group, and the levels of IL-4 and IL-10 showed an upward trend compared with the APP group.

[0176] CCL-2 is involved in the formation of inflammatory response. When it is used to treat microglia, the latter's migration increases abnormally. Microglia can activate this factor, thereby mediating immune damage and inflammatory response of neural tissue cells. CXCL1 belongs to the CXC chemokine family and is widely present in the nervous system. It is one of the important inflammatory mediators of nervous system lesions such as brain and spinal cord trauma and neurodegenerative diseases. The main function of the anti-inflammatory factor IL-4 is to promote humoral immunity and inhibit cellular immunity, thereby enhancing the killing effect of cells. The main function of the anti-inflammatory factor IL-10 is to inhibit immune response, inhibit inflammatory response, and inhibit cell apoptosis. Therefore, the above results show that AKK PROBIO significantly upregulated the expression of anti-inflammatory factors IL-4 and IL-10, significantly downregulated the levels of pro-inflammatory factors CXCL1 and CCL2, and improved the inflammatory response of the nervous system by regulating inflammatory factors.

[0177] Compared with the model group, the IL-4 level in the APP+AKK PROBIO group increased by about 2.5 times, the IL-10 level increased by about 1.4 times, the CXCL1 level decreased by about 51%, the CCL2 level decreased by about 25%, and the Aβ level in the hippocampus decreased by about 50% ( Figure 3 ).

[0178] Table 2 The results of the determination of the inflammatory indexes in mice in this example

[0179]

[0180] *The same lowercase letters indicate no significant difference between the two groups, and different lowercase letters indicate significant difference between the two groups (p<0.05); pg in pg / mL is the mass of IL-4, IL-10, CXCL1 or CCL2, and mL is the volume of serum; ng / mL is the mass of Aβ, and mL is the volume of serum.

[0181] 2.4 Determination of the effect of AKK PROBIO on the expression of inflammatory factor-related genes in hippocampal tissue

[0182] Take the hippocampal tissue of each group of mice in this embodiment, and detect the mRNA expression of IL-1β, Caspase-1, IL-18, IDE and NLRP3 in the hippocampal tissue of mice by qPCR. Take the hippocampal tissue of mice in a homogenization tube containing homogenization beads, add 1mL TRIzol reagent, and homogenize. Add 200μL chloroform after homogenization, mix well and let stand on ice for 15min. Centrifuge and take the supernatant in a new centrifuge tube, add isopropanol and mix well, let stand for 20min, centrifuge at 14000r / min for 20min at 4°C, the white precipitate at the bottom of the tube is RNA, pour out the supernatant, add 75% ethanol for cleaning, centrifuge at 14000r / min for 15min at 4°C, extract the liquid in the tube, place it on the clean bench for 3min drying, and then add 20μL DEPC water to dissolve the RNA, which is the RNA stock solution. RNA is transcribed into cDNA using a transcription kit. 10 μL SYBR Green PCR Master Mix, 4 μL sterile ultrapure water, 2 μL primers, and 2 μL cDNA were added to the eight tubes in sequence. Real-time PCR was performed for 40 cycles of pre-denaturation at 95°C for 10 min, denaturation at 95°C for 15 min, annealing at 60°C for 30 s, and extension at 60°C for 30 s. GAPDH was used as the housekeeping gene and 2- ΔΔCt The expression level of target gene mRNA was calculated by the method. The primer sequences used are shown in Table 3. The detection results are shown in Table 4.

[0183] Table 3 Primers for detecting the expression of genes related to inflammatory factors

[0184]

[0185]

[0186] Table 4 Results of determination of inflammatory factor-related gene expression

[0187]

[0188] *The same lowercase letters indicate no significant difference between the two groups, and different lowercase letters indicate significant difference between the two groups (p<0.05).

[0189] The results showed that the expression of inflammatory factors IL-1β, IL-18 and NLRP3, and apoptosis factor Caspase-1 in the APP group was significantly increased, which was about 3, 4, 6 and 4 times higher than that in the normal group, respectively. In addition, the Aβ-degrading enzyme IDE was expressed at the lowest level in the APP group, which was downregulated by about 79% compared with the normal group. After AKK PROBIO intervention, the activity expression of IDE was significantly enhanced, which was about 2 times higher than that in the model group (APP group). Compared with the APP group, the APP+AKK PROBIO group significantly downregulated the expression of IL-1β, IL-18, NLRP3 and Caspase-1, among which the level of IL-1β was downregulated by about 46%, the level of IL-18 was downregulated by 44%, the level of NLRP3 was downregulated by about 40%, and the level of Caspase-1 was downregulated by about 47%.

[0190] 2.5 Effect of AKK PROBIO on Aβ protein expression in hippocampus

[0191] The hippocampal tissue of each group of mice in this example was taken, and the hippocampal tissue was homogenized in 1 mL RIPA (Thermofisher, Waltham, MA, USA) and 10 μL PMSF (Thermofisher), and centrifuged at 12000×g for 5 min at 4°C. The protein was quantified using a BCA protein assay kit (Thermofisher). The protein sample was mixed with a sample buffer (Thermofisher) at a ratio of 4:1 and heated at 95°C for 5 min, and then the sample was spotted into the wells of the SDS-PAGE gel and run at 100 V. The bands on the SDS-PAGE gel were transferred to a PVFD membrane, the PVDF membrane was blocked with 5% skim milk for 1 h, and then incubated with a primary antibody (Thermofisher) at 4°C overnight, and the antibody (Thermofisher) was added after washing the membrane and incubated for 1 hour. Images were obtained on iBright (Thermofisher) after chemiluminescence using a western ECL substrate (Thermofisher) for subsequent analysis.

[0192] The results showed that the expression of Aβ protein in the APP+AKK PROBIO group was significantly downregulated compared with the APP group, but still higher than that in the WT group ( Figure 4 ).

[0193] Example 3 Effects of AKK PROBIO on Mouse Liver and Colon Tissues

[0194] The feeding, modeling and grouping of mice in this example are the same as those in Example 2.

[0195] The livers of mice in each group were taken, and liver pathological sections were made and observed. The results showed that the liver lobules of mice in each group had orderly structures, and the central veins and hepatic sinusoids were clear ( Figure 5 ). This indicates that the administration of AKK PROBIO is non-toxic to the liver of mice and has a good safety profile.

[0196] The intestinal tissue of mice was sliced ​​and observed. The results showed that the intestinal tissue structure of WT mice was intact and normal in morphology. In the APP group, the intestinal muscular layer cells of the mice were wrinkled, the crypt cells were damaged, the mucosal tissue was atrophied, and the fold inflammation was enriched. In the APP+AKK PROBIO group, the intestinal muscle cells recovered, and the crypt and mucosal tissues tended to be normal ( Figure 6 ). This indicated that the inflammation enrichment in the APP+AKKPROBIO group was less than that in the APP group.

[0197] Comparative Example

[0198] In this comparative example, common AKK bacteria (purchased from ATCC, CAT#: ATCC-BAA835) was selected as the control group to compare the difference between AKK PROBIO and AKK PROBIO in improving spatial memory ability.

[0199] The experiment of this comparative example was carried out in parallel for three tissue samples, and then the mean value was calculated. SPSS software (SPSS v.25 for Windows, IBM Software Group, Chicago, IL, USA) was used to average and analyze the data. The differences between the mean values ​​of each group were evaluated by one-way analysis of variance using Duncan's multiple range test. The difference of p<0.05 was considered statistically significant.

[0200] The mouse model was constructed as in Example 2.

[0201] Experimental groups: 30 12-month-old APP / PS1 double transgenic mice were randomly divided into APP group (model group), APP+AKKPROBIO group (administered AKK PROBIO to AD model mice), and control group (administered ATCC-BAA835 bacteria to AD model mice). Ten wild-type C57 BL / 6J mice from the same litter were used as blanks. The WT group was gavaged with normal saline every day, the APP group was gavaged with normal saline every day, and the APP+AKK PROBIO group was gavaged with 1×10 9 CFU / day / AKK PROBIO solution in normal saline, the control group was gavaged daily at a concentration of 1×10 9 CFU / day / ATCC-BAA835 solution in normal saline.

[0202] The water maze experimental conditions were the same as in Example 2.

[0203] The results are shown in Table 5. After 4 consecutive days of Morris water maze test, the escape latency of mice in each group showed a decreasing trend. Compared with the test results of the APP group on the last day, the escape latency of mice in the APP+AKK PROBIO group was shortened by about 24%. The escape latency of mice in the APP+ATCC-BAA835 group was shortened by only 10.3%. This shows that the AKKPROBIO bacteria provided by the present invention has a stronger ability to improve spatial memory than ordinary Akkermansia muciniphila.

[0204] Table 5 Escape latency test results of mice in each group (seconds, s)

[0205]

[0206]

[0207] *The same lowercase letters indicate no significant difference between the two groups, and different lowercase letters indicate significant difference between the two groups (p<0.05).

[0208] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. Akkermansia muciniphila Use in the preparation of a product for the prevention, treatment and / or auxiliary treatment of Alzheimer's disease, characterized in that: Said Akkermansia muciniphila It is deposited in CGMCC with the deposit number CGMCC No.20955; the product is a medicine and pharmaceutical raw material.

2. The use according to claim 1, characterized in that: The products include Akkermansia muciniphila form of survival.

3. The use according to claim 1, characterized in that: The drug further comprises at least one of a pharmaceutically acceptable excipient, a pharmaceutically acceptable compound, a pharmaceutically acceptable protein and / or a pharmaceutically acceptable polypeptide.

4. The use according to claim 3, characterized in that: The pharmaceutically acceptable excipient is selected from at least one of solvents, diluents, disintegrants, precipitation inhibitors, surfactants, glidants, binders, lubricants, dispersants, suspending agents, isotonic agents, thickeners, emulsifiers, preservatives, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, flavoring agents, sweeteners, ion exchangers, release agents, coating agents, flavoring agents and antioxidants.

5. The use according to claim 1, characterized in that: The dosage form of the drug is selected from at least one of powders, tablets, granules, capsules, solutions, emulsions, suspensions, injections, sprays, powder sprays, aerosols, suppositories, drops and pills.

6. The use according to claim 1, characterized in that: Each unit of the drug contains at least Akkermansia muciniphila 10 8 CFU / g or CFU / mL.

7. The use according to claim 1, characterized in that: Each unit of the drug contains Akkermansia muciniphila 10 8 -10 12 CFU / g or CFU / mL.

8. The use according to claim 7, characterized in that: Each unit of the drug contains Akkermansia muciniphila 10 9 -10 12 CFU / g or CFU / mL.

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