Probiotic formulation for improving parkinson's disease and use thereof

Abstract

The present application relates to a kind of probiotic preparation for improving Parkinson's disease and its application, the strain in the probiotic preparation includes mucinophilic Akkermansia Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve BBr60 strain.The present application develops a brand-new probiotic compound mode and a brand-new probiotic intervention strategy for improving Parkinson's disease, i.e., Akk11 strain and BBr60 strain are compounded, it is found that there is potential interaction between the two, can cooperate with each other, synergistic effect in improving the efficacy of Parkinson's disease, specifically in: synergistically improving the motor disorders caused by Parkinson's disease, reducing the expression of related inflammatory factors in substantia nigra, promoting the generation of anti-inflammatory factors to relieve neurogenic inflammation, inhibiting the activation of inflammasome NLPR3 in substantia nigra to further inhibit brain inflammation.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of probiotic preparations, and relates to a probiotic preparation for improving Parkinson's disease and application thereof. BACKGROUND

[0002] The main pathological features of Parkinson's disease are the degeneration of dopaminergic neurons in the substantia nigra of the midbrain, the blockage of nerve conduction, and the formation of Lewy bodies caused by the abnormal aggregation of alpha-synuclein in neurons. The clinical features include typical motor dysfunction, such as tremor, muscle stiffness, slowness of movement, and gait instability, as well as various non-motor symptoms, which seriously affect the quality of life of patients and their families.

[0003] The treatment of Parkinson's disease mainly focuses on relieving symptoms and improving the quality of life. Through drug, surgery and lifestyle intervention, the disease can be effectively controlled. Common drug treatments include levodopa, dopamine agonists, and anticholinergic drugs, which mainly relieve motor symptoms by supplementing dopamine or adjusting neurotransmitters. For patients who do not respond well to drug treatment, surgical treatments such as deep brain stimulation (DBS) are used to improve motor symptoms. In addition, some auxiliary treatments such as physical therapy, occupational therapy, and speech therapy are also used to maintain the daily life of patients.

[0004] Current treatments for Parkinson's disease can relieve symptoms, but each has its own drawbacks. Drug treatments such as levodopa can cause motor complications and drug resistance, dopamine agonists can cause drowsiness and impulse control disorders, and anticholinergic drugs can cause side effects such as cognitive impairment. Surgical treatments such as deep brain stimulation have surgical risks and require device adjustments, and intracerebral injection of drugs is expensive and has varying effects. Auxiliary treatments can improve function, but they require a long treatment time and the effects vary from person to person. In short, these methods have certain side effects and limitations while improving symptoms.

[0005] Gut microbiota may play an important role in the occurrence and progression of Parkinson's disease. Parkinson's disease patients often have gastrointestinal dysfunction, such as difficulty swallowing, nausea, vomiting, and constipation. Inflammatory cytokines in the gut, such as IL-1β, can cross the blood-brain barrier into the brain, further causing microglial cells in the brain to transform into M1-activated state, mediating and triggering neuroinflammatory responses to release a large amount of inflammatory cytokines such as IL-1β and IL-18, as well as reactive oxygen species (ROS), thereby exacerbating the pathological process of Parkinson's disease. Moreover, Parkinson's disease patients often have certain intestinal flora imbalance. Intestinal microbiota imbalance has been observed in both Parkinson's disease patients and Parkinson's disease model mice, with an increase in harmful bacteria such as Escherichia coli and Shigella, and a decrease in beneficial bacteria such as Bifidobacterium and Akkermansia.

[0006] Probiotics can not only regulate immunity, but also reduce inflammation, possibly through the "gut-brain axis" to interact with the brain, enhance vagus nerve function, improve neuroinflammation, and thus have a positive impact on neurodegenerative diseases such as Parkinson's disease. Compared with traditional Parkinson's disease treatment methods, the advantages of probiotic therapy are less side effects, and less likely to cause drug resistance or dependence. As a non-invasive treatment, probiotics have a wide range of applications, can be used for a long time, and are less likely to be limited by the patient's physical condition. Therefore, it is very meaningful to develop more probiotic intervention strategies to improve Parkinson's disease. SUMMARY

[0007] In view of the deficiencies of the prior art, the purpose of the present application is to provide a probiotic preparation for improving Parkinson's disease and its application.

[0008] To achieve the purpose of the present application, the following technical solutions are adopted:

[0009] In a first aspect, the present application provides a probiotic preparation for improving Parkinson's disease, wherein the strains in the probiotic preparation include mucinophilic Akkermansia muciniphila with a preservation number of CCTCC NO: M2024119 Akkermansia muciniphila Akk11 strain and Bifidobacterium breve with a preservation number of CGMCC No.12915 Bifidobacterium breve BBr60 strain.

[0010] The present application develops a brand-new probiotic compounding method and a brand-new probiotic intervention strategy for improving Parkinson's disease, which is to compound mucinophilic Akkermansia muciniphila Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve BBr60 strain, and it is found that the two strains have potential interaction and can cooperate with each other to synergistically improve the efficacy of Parkinson's disease, which is specifically manifested in: (1) synergistically improving the motor disorders caused by Parkinson's disease; (2) synergistically reducing the expression of related inflammatory factors in the substantia nigra and synergistically promoting the generation of anti-inflammatory factors to relieve neurogenic inflammation; (3) synergistically inhibiting the activation of inflammasome NLPR3 in the substantia nigra to further inhibit brain inflammation. At the same time, as probiotics, mucinophilic Akkermansia muciniphila Akk11 and Bifidobacterium breve BBr60 have high product safety and are less likely to produce resistance.

[0011] Preferably, the ratio of viable bacteria of the mucinophilic Akkermansia muciniphila Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve BBr60 strain is 1:3-2:1, such as 1:3, 2:5, 1:2, 2:3, 1:1, 3:2, 2:1, and other specific point values in the numerical range can be selected, which will not be described one by one here.

[0012] Based on the potential synergistic relationship between the two strains, when the specific viable cell count ratio is met, the effect of improving motor disorders caused by Parkinson's disease, reducing the expression of related inflammatory factors in the substantia nigra, promoting the generation of anti-inflammatory factors, and inhibiting the activation of inflammasome NLPR3 in the substantia nigra is more excellent.

[0013] Preferably, in the probiotic preparation, the total number of viable bacteria is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, for example, 1×10 9 CFU / g (CFU / mL), 2×10 9 CFU / g (CFU / mL), 5×10 9 CFU / g (CFU / mL), 8×10 9 CFU / g (CFU / mL), 1×10 10 CFU / g (CFU / mL), 5×10 10 CFU / g (CFU / mL), 1×10 11 CFU / g (CFU / mL), 1×10 12 CFU / g (CFU / mL), 1×10 13 CFU / g (CFU / mL), etc. Other specific point values within the numerical range can be selected, which will not be repeated here.

[0014] Preferably, the dosage form of the probiotic preparation includes a solution, a powder, a capsule, a tablet, or a granule. The dosage form of the probiotic preparation involved in the present application is not limited, including the most commonly used solution, powder, or further prepared capsule, tablet, or granule.

[0015] Preferably, the dosage form of the probiotic preparation is a solution, which is prepared by the following method:

[0016] The Akk11 strain and the BBr60 strain are inoculated into the culture medium respectively for activation and fermentation culture in turn to obtain fermentation broth; the fermentation broth is centrifuged respectively, resuspended with a solvent to obtain Akk11 bacterial suspension and BBr60 bacterial suspension; the Akk11 bacterial suspension and the BBr60 bacterial suspension are mixed according to the viable cell count ratio, and the probiotic preparation is obtained.

[0017] Preferably, the dosage form of the probiotic preparation is a powder, which is prepared by the following method:

[0018] The Akk11 strain and the BBr60 strain are inoculated into culture media respectively to carry out activation and fermentation culture in turn to obtain fermentation liquor; the fermentation liquor is centrifuged, mixed with a protective agent and then freeze-dried to obtain Akk11 bacterial powder and BBr60 bacterial powder; the Akk11 bacterial powder and the BBr60 bacterial powder are mixed according to the ratio of viable bacterial numbers, and the probiotic preparation is obtained.

[0019] Preferably, the probiotic preparation further contains an auxiliary material; the auxiliary material includes any one or a combination of at least two of a filler, a binder, a wetting agent, a disintegrating agent, an emulsifying agent, a solubilizing agent, a penetration pressure regulator, a coloring agent, a pH regulator, an antioxidant, a bacteriostatic agent or a buffer.

[0020] In the present application, the probiotic preparation improves motor dysfunction.

[0021] In the present application, the probiotic preparation reduces the expression level of inflammatory factors.

[0022] In the present application, the probiotic preparation inhibits the activation of NLRP3 inflammasome.

[0023] In a second aspect, the present application provides a probiotic preparation for assisting in improving Parkinson's disease, wherein the strain in the probiotic preparation includes a mucinophilic Akkermansia muciniphila with a preservation number of CCTCC NO: M2024119. Akkermansia muciniphila Akk11 strain and Bifidobacterium breve with a preservation number of CGMCC No.12915 Bifidobacterium breve BBr60 strain.

[0024] Preferably, the mucinophilic Akkermansia muciniphila Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve The ratio of viable bacterial numbers of the Akk11 strain and the BBr60 strain is 1:3-2:1, for example, 1:3, 2:5, 1:2, 2:3, 1:1, 3:2, 2:1, and other specific point values in the numerical range can be selected, which will not be described here one by one.

[0025] Preferably, in the probiotic preparation, the total number of viable bacteria is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g, for example, 1×10 9 CFU / g (CFU / mL), 2×10 9 CFU / g (CFU / mL), 5×10 9 CFU / g (CFU / mL), 8×10 9 CFU / g (CFU / mL), 1×10 10CFU / g (CFU / mL), 5x10 10 CFU / g (CFU / mL), 1x10 11 CFU / g (CFU / mL), 1x10 12 CFU / g (CFU / mL), 1x10 13 CFU / g (CFU / mL), 1x10

[0026] In a third aspect, the present application provides use of the probiotic preparation for improving Parkinson's disease according to the first aspect in preparation of an inhibitor of NLRP3 inflammasome activation.

[0027] Compared with the prior art, the present application has the following beneficial effects:

[0028] The present application develops a brand-new probiotic compound mode and a brand-new probiotic intervention strategy for improving Parkinson's disease, which is to compound Akkermansia muciniphila Akk11 and Bifidobacterium breve BBr60. Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve BBr60 strain, and it is found that the two strains have potential interaction and can cooperate with each other to synergistically improve the efficacy of Parkinson's disease, which is specifically shown in the following aspects: (1) synergistically improving the motor disorder caused by Parkinson's disease; (2) synergistically reducing the expression of related inflammatory factors in the substantia nigra and synergistically promoting the generation of anti-inflammatory factors to relieve neurogenic inflammation; (3) synergistically inhibiting the activation of inflammasome NLPR3 in the substantia nigra to further inhibit brain inflammation. At the same time, as probiotics, Akkermansia muciniphila Akk11 and Bifidobacterium breve BBr60 have high safety and are not prone to resistance.

[0029] The classification name of the Akk11 strain involved in the present application is Akkermansia muciniphila Akk11 Akkermansia muciniphila , which is preserved in China Center for Type Culture Collection, preserved on January 15, 2024, has a preservation number of CCTCC NO: M2024119, and is located at Wuhan University, Wuhan, China.

[0030] The classification name of the BBr60 strain involved in the present application is Bifidobacterium breve Bifidobacterium breve , which is preserved in China General Microbiological Culture Collection Center, preserved on August 29, 2016, has a preservation number of CGMCC No.12915, and is located at No.3, Beichen West Road, Beijing, China. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 is a statistical result graph of the pole falling time of each group of mice in the pole climbing test;

[0032] Figure 2 This is a statistical graph showing the residence time of each group of mice in the rotarod test;

[0033] Figure 3 This is the statistical result of the total moving distance of each group of mice in the central area in the open field test;

[0034] Figure 4 This is a statistical graph of the number of times each group of mice entered the central area in the open field test;

[0035] Figure 5 This is a statistical result diagram of the relative expression of the pro-inflammatory factor IL-1β gene in each group of mice in the inflammatory factor expression test;

[0036] Figure 6 This is a statistical result chart of the relative expression of the pro-inflammatory factor TNF-α gene in each group of mice in the inflammatory factor expression test;

[0037] Figure 7 This is a statistical result chart of the relative expression of the pro-inflammatory factor IL-6 gene in each group of mice in the inflammatory factor expression test;

[0038] Figure 8 This is a statistical result chart of the relative expression of the anti-inflammatory factor TGF-β gene in each group of mice in the inflammatory factor expression test;

[0039] Figure 9 This is the statistical result of the relative expression of the anti-inflammatory factor IL-10 gene in each group of mice in the inflammatory factor expression test;

[0040] Figure 10 This is the statistical result of the relative expression of the anti-inflammatory factor Arg-1 gene in each group of mice in the inflammatory factor expression test;

[0041] Figure 11 This is the statistical result of the relative expression of the inflammasome-related protein NLRP3 in the substantia nigra tissue of each group of mice;

[0042] Figure 12 This is a statistical result of the relative expression of inflammasome-related protein caspase-1 in the substantia nigra tissue of each group of mice;

[0043] Figure 13 This is the statistical result of the relative expression of inflammasome-related protein IL-1β in the substantia nigra tissue of each group of mice;

[0044] In the figure, ns indicates no significant difference compared with the model group, # indicates p < 0.05 compared with the model group, ## indicates p < 0.01 compared with the model group, ### indicates p < 0.001 compared with the model group, #### indicates p < 0.0001 compared with the model group, and **** indicates p < 0.0001 compared with the NC group. DETAILED DESCRIPTION

[0045] The technical solutions of the present application are further illustrated by specific embodiments. Those skilled in the art should understand that the embodiments are only used to help understand the present application and should not be regarded as specific limitations of the present application.

[0046] The classification name of the Akk11 strain involved in the following content is Akkermansia muciniphila Akk11 Akkermansia muciniphila Akk11, deposited on January 15, 2024, with the accession number CCTCC NO: M2024119.

[0047] The classification name of the BBr60 strain involved in the following content is Bifidobacterium breve Bifidobacterium breve , deposited on August 29, 2016, with the accession number CGMCC No. 12915.

[0048] The method for preparing the bacterial suspension involved in the following test is as follows: inoculate the desired strain into MRS liquid medium, cultivate at 37°C for 20 h for activation, continuously activate for 2 times, and obtain an activation liquid; inoculate the activation liquid into MRS liquid medium, cultivate at 37°C, and obtain a bacterial liquid; centrifuge the bacterial liquid, filter, obtain bacterial bodies, resuspend the bacterial bodies with PBS, and obtain the bacterial suspension.

[0049] MRS medium: 10 g / L proteose peptone, 10 g / L beef extract, 20 g / L glucose, 2 g / L sodium acetate, 5 g / L yeast powder, 2 g / L diammonium hydrogen citrate, 2.6 g / L K2PO4·3H2O, 0.1 g / L MgSO4·7H2O, 0.05 g / L MnSO4, 80 mL / L Tween 80, and 0.5 g / L cysteine hydrochloride.

[0050] Example

[0051] This example discusses the improvement effect of probiotic preparation on a Parkinson mouse model:

[0052] (1) Test animals:

[0053] SPF male C57BL / 6J mice (5 weeks old, purchased from Jiangsu Jizhuangkang Biotechnology Co., Ltd.) were first orally administered with 200 μL cocktail antibiotics (1 g / L metronidazole, 0.5 g / L vancomycin, 1 g / L ampicillin, and 1 g / L neomycin) for 7 consecutive days. The mice were raised in a room temperature (22±1°C), humidity (55±5%), and pathogen-free environment (12 hours, light / dark cycle), and the animals were free to eat and drink.

[0054] (2) Animal grouping:

[0055] The mice were adaptively fed for 1-7 days in the experiment, and after the mice were adaptively fed for one week, they were randomly assigned into 8 groups of 8 mice each: an untreated group (NC group), an MPTP model group, an Akk11 group (model mice were intervened with Akk11 bacterial solution), a BBr60 group (model mice were intervened with BBr60 bacterial solution), a compound 1 group (model mice were intervened with Akk11 bacterial solution and BBr60 bacterial solution in combination, with a ratio of live bacteria of 1:3), a compound 2 group (model mice were intervened with Akk11 bacterial solution and BBr60 bacterial solution in combination, with a ratio of live bacteria of 2:1), a compound control group (model mice were intervened with Akk11 bacterial solution and commercially available Bifidobacterium breve ATCC15700 bacterial solution in combination, with a ratio of live bacteria of 1:3), and a probiotic control group (healthy mice were intervened with Akk11 bacterial solution and BBr60 bacterial solution in combination, with a ratio of live bacteria of 1:3).

[0056] (3) Animal modeling and intervention methods:

[0057] Untreated group: from the 7th day to the 35th day of the experiment, the mice were given biological saline by gavage every day;

[0058] MPTP model group: from the 14th day to the 19th day of the experiment, the mice were given intraperitoneal injection of MPTP (30 mg / kg) and probenecid (250 mg / kg) every day, and at the same time, from the 7th day to the 35th day of the experiment, the mice were given biological saline by gavage every day;

[0059] Akk11 group: from the 14th day to the 19th day of the experiment, the mice were given intraperitoneal injection of MPTP (30 mg / kg) and probenecid (250 mg / kg) every day, and at the same time, from the 7th day to the 35th day of the experiment, the mice were given Akk11 bacterial suspension (1×10 9 CFU per day per mouse) by gavage every day;

[0060] BBr60 group: from the 14th day to the 19th day of the experiment, the mice were given intraperitoneal injection of MPTP (30 mg / kg) and probenecid (250 mg / kg) every day, and at the same time, from the 7th day to the 35th day of the experiment, the mice were given BBr60 bacterial suspension (1×10 9 CFU per day per mouse) by gavage every day;

[0061] Compound 1 group: from the 14th day to the 19th day of the experiment, the mice were given intraperitoneal injection of MPTP (30 mg / kg) and probenecid (250 mg / kg) every day, and at the same time, from the 7th day to the 35th day of the experiment, the mice were given Akk11 and BBr60 mixed bacterial suspension (total intervention amount 1×10 9 CFU per day per mouse, with a ratio of live bacteria of 1:3) by gavage every day;

[0062] Compound 2 group: from day 14 to day 19 of the experiment, mice were given intraperitoneal injection of MPTP (30 mg / kg) and probenecid (250 mg / kg) every day, while from day 7 to day 35 of the experiment, mice were given gavage of Akk11 and BBr60 mixed bacteria suspension (total intervention amount 1 x 10 9 CFU / day / animal, the ratio of viable bacteria was 2:1);

[0063] Compound control group: from day 14 to day 19 of the experiment, mice were given intraperitoneal injection of MPTP (30 mg / kg) and probenecid (250 mg / kg) every day, while from day 7 to day 35 of the experiment, mice were given gavage of Akk11 and ATCC15700 mixed bacteria suspension (total intervention amount 1 x 10 9 CFU / day / animal, the ratio of viable bacteria was 1:3);

[0064] Probiotic control group: from day 7 to day 35 of the experiment, mice were given gavage of Akk11 and BBr60 mixed bacteria suspension (total intervention amount 1 x 10 9 CFU / day / animal, the ratio of viable bacteria was 1:3).

[0065] (4) Exercise behavior test:

[0066] (4.1) Pole climbing test:

[0067] A vertical metal rod, about 50 cm in length and 1 cm in diameter, was prepared. Two days before the test, all mice received adaptive training to minimize the impact of environmental changes on the test results. Specifically, the mice were placed in the rod cage to familiarize themselves with the environment, and then placed head-up on the rod 15 cm from the cage bottom for 3 times, and then placed on the rod 30 cm and 50 cm above the ground of the cage for 3 times of training. On the test day, each mouse was placed head-up on the top of the rod, and the process of descending was observed, and the time until the two forepaws touched the ground was recorded. The test was repeated 3 times with an interval of 5 min, and if the mouse fell or jumped off the rod within 60 s, the test was repeated, and the average time was taken for statistical analysis. The results are shown in Figure 1 .

[0068] (4.2) Rotarod test:

[0069] The RT-01 mouse rotarod containing an automatic timer and a falling sensor was used during training and formal testing. Before the formal test, the mice received three training sessions at three incremental speeds (10, 20, and 30 rpm) for 180 s per day for three days. During the formal experiment, the rotating rod was set to rotate at a speed of 30 rpm, and the mice were placed on the rotating rod, and the latency to fall was recorded, and the average time was taken. The results are shown in Figure 2 .

[0070] (4.3) Open field test:

[0071] Two days before the test, the mice were subjected to environmental adaptation training to reduce the stress caused by the new environment in the formal experiment. The open field consisted of a 50 cm x 50 cm square arena with four walls, each 45 cm high. During the formal test, the laboratory was kept quiet and properly lit, with minimal human intervention. The mice were gently placed in the center of the empty field, and their behavior was recorded for 5 min, with the total distance and number of entries into the center recorded. The results are shown in Figure 3 and Figure 4 .

[0072] As can be seen from Figures 1-4 , compared with the NC group, the model group mice had significantly longer time on the descending rod, significantly shorter time falling on the rotating rod, and poorer performance in total distance and number of entries into the open field. After intervention with probiotics in each group, the time on the descending rod was shortened to varying degrees, the time falling on the rotating rod was extended to varying degrees, and the total distance and number of entries into the open field were increased to varying degrees. The improvement in the above aspects was more obvious in the composite 1 and composite 2 groups than in the Akk11 and BBr60 groups, indicating that the Akk11 and BBr60 strains cooperate and promote each other in improving the motor dysfunction of Parkinson's mice.

[0073] (5) Inflammatory factor expression test:

[0074] After the motor behavior test, the mice were sacrificed and their tissues were collected for analysis. Total RNA was extracted from the mouse brain substantia nigra tissue using a kit. Reverse transcription was then performed using a kit, and finally qRT-PCR analysis was performed. The expression of related genes was normalized to the level of β-actin, and the results are shown in Figures 5-10 .

[0075] As can be seen from Figures 5-10 , compared with the NC group, the pro-inflammatory cytokines (IL-1β, TNF-α, IL-6) of the model group mice were significantly increased, while the anti-inflammatory cytokines (TGF-β, IL-10, Arg-1) were decreased. After intervention with probiotics in each group, the levels of pro-inflammatory factors were decreased to varying degrees, and the levels of anti-inflammatory factors were increased to varying degrees. The improvement in the above aspects was more obvious in the composite 1 and composite 2 groups than in the Akk11 and BBr60 groups, indicating that the Akk11 and BBr60 strains cooperate and promote each other in reducing the inflammatory response of Parkinson's mice.

[0076] (6) Inflammasome protein immunoblotting test:

[0077] The substantia nigra tissues of each group of mice were lysed using RIPA lysis buffer containing a protease inhibitor cocktail. The protein samples were separated by SDS-PAGE and electrotransferred to a polyvinylidene fluoride membrane. The membrane was blocked with 5% skim milk at room temperature for 2 hours. Then, the primary antibody of the following antibodies was incubated overnight: β-actin (1:3000), NLRP3 (1:1000), Caspase-1 (1:1000), IL-1β (1:1000), incubated at 4°C overnight. Then, the membrane was incubated with the secondary antibody of anti-mouse IRDye® 680RD conjugated antibody and anti-rabbit IRDye® 800CW conjugated antibody. The immunoblot was scanned using a dual-color infrared laser imager, and analyzed using software. The statistical results of the relative amount of protein expression of each group are shown in Table 2. Figures 11-13

[0078] NLRP3 inflammasome plays an important role in the pathogenesis of Parkinson's disease, mainly through microglial cell activation, pro-inflammatory factor release, mitochondrial damage and cell pyroptosis to accelerate the death of dopaminergic neurons. Figure 7 The results show that, compared with the NC group, the levels of inflammation-related proteins NLRP3, Caspase-1 / pro-Caspase-1 and IL-1β / pro-IL-1β in the model group of mice are significantly increased; the expression trend of these proteins is reversed in each group of probiotics intervention, and the above-mentioned reverse effect of the compound 1 group and the compound 2 group is more obvious than that of the Akk11 group and the BBr60 group, indicating that the Akk11 strain and the BBr60 strain cooperate and promote each other in inhibiting the activation of NLRP3 inflammasome in the substantia nigra tissue of Parkinson's mice.

[0079] The applicant declares that the technical solutions of the present application are illustrated by the above-mentioned embodiments, but the present application is not limited to the above-mentioned embodiments, i.e. it does not mean that the present application must rely on the above-mentioned embodiments to be implemented. It should be understood by those skilled in the art that any improvement of the present application, equivalent replacement of each raw material of the product of the present application and addition of auxiliary ingredients, selection of specific modes, etc. fall within the protection scope and disclosure scope of the present application.

[0080] The preferred embodiments of the present application are described in detail above, but the present application is not limited to the specific details in the above-mentioned embodiments, and various simple modifications can be made to the technical solutions of the present application within the technical concept of the present application, and these simple modifications all belong to the protection scope of the present application.

[0081] In addition, it should be noted that each specific technical feature described in the above-mentioned specific embodiments can be combined in any appropriate manner without contradiction, and in order to avoid unnecessary repetition, the present application will not further describe various possible combination manners.​

Claims

1. A probiotic preparation for improving Parkinson's disease, characterized in that: The strains in the probiotic preparation are composed of Akkermansia muciniphila Akk11 strain with a deposit number of CCTCC NO: M2024119 and Bifidobacterium breve BBr60 strain with a deposit number of CGMCC No.12915; The ratio of the viable bacteria count of the Akkermansia muciniphila Akk11 strain to the Bifidobacterium breve BBr60 strain is 1:3-2:

1.

2. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that In the probiotic preparation, the total number of viable bacteria is not less than 1×10 9 CFU / mL or 1×10 9 CFU / g.

3. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that The dosage form of the probiotic preparation includes solution, powder, capsule, tablet or granule.

4. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that The dosage form of the probiotic preparation is a solution, which is prepared by the following method: The Akk11 strain and the BBr60 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain fermentation broth; the fermentation broth is centrifuged respectively and resuspended with a solvent to obtain an Akk11 bacterial suspension and a BBr60 bacterial suspension; the Akk11 bacterial suspension and the BBr60 bacterial suspension are mixed according to the ratio of the number of viable bacteria to obtain.

5. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that The probiotic preparation is in the form of a powder, which is prepared by the following method: The Akk11 strain and the BBr60 strain are inoculated into a culture medium respectively for activation and fermentation culture in sequence to obtain fermentation broth; the fermentation broth is centrifuged respectively, mixed with a protective agent and then freeze-dried to obtain Akk11 bacterial powder and BBr60 bacterial powder; the Akk11 bacterial powder and the BBr60 bacterial powder are mixed according to the ratio of the number of viable bacteria to obtain.

6. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that The probiotic preparation further contains excipients; the excipients include any one of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, antibacterial agents or buffers, or a combination of at least two of them.

7. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that The probiotic preparation improves motor dysfunction.

8. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that The probiotic preparation reduces the expression level of inflammatory factors and inhibits the activation of NLRP3 inflammasome.

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