Probiotic preparation for improving Parkinson's disease and application thereof

Abstract

The invention relates to a probiotic preparation for improving Parkinson's disease and an application of the probiotic preparation. The bacterial strains in the probiotic preparation comprise an Ackermania muciniphila Akk11 bacterial strain and a Bifidobacterium breve BBr60 bacterial strain. The invention further relates to a preparation method of the probiotic preparation for improving the Parkinson's disease and an application of the probiotic preparation for improving the Parkinson's disease, wherein the bacterial strains in the probiotic preparation comprise the Ackermania muciniphila Akk11 bacterial strain and the Bifidobacterium breve BBr60 bacterial strain. The invention develops a brand-new probiotic compounding mode and a brand-new probiotic intervention strategy for improving the Parkinson's disease, that is, the Akk11 strain and the BBr60 strain are compounded, and the two strains are found to have potential interaction and can be matched with each other, so that a synergistic effect is achieved on the aspect of improving the Parkinson's disease, and the effect of improving the Parkinson's disease is achieved. Specifically, the traditional Chinese medicine composition synergistically improves dyskinesia caused by Parkinson's disease, reduces expression of related inflammatory factors in nigra, promotes generation of anti-inflammatory factors to relieve neurological inflammation, inhibits activation of inflammasome NLPR3 in nigra, and further inhibits brain inflammation.

Description

Technical Field

[0001] The present invention belongs to the technical field of probiotic preparations and relates to a probiotic preparation for improving Parkinson's disease and an application thereof. Background Art

[0002] The primary pathological hallmarks of Parkinson's disease are degeneration of dopaminergic neurons in the substantia nigra of the midbrain, impaired nerve conduction, and the abnormal aggregation of α-synuclein within neurons, forming Lewy bodies. Clinical features include typical motor dysfunction such as tremor, muscle rigidity, bradykinesia, and gait instability, as well as a variety of non-motor symptoms, which severely impact the quality of life of patients and their families.

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

[0004] While current treatments for Parkinson's disease can alleviate symptoms, they each have drawbacks. Drug treatments such as levodopa can cause motor complications and drug resistance, dopamine agonists can cause drowsiness and impulse control disorders, and anticholinergics can cause side effects such as cognitive impairment. Surgical treatments such as deep brain stimulation carry surgical risks and require equipment adjustments, while intracerebral drug injections are expensive and have varying effectiveness. While adjunctive therapies can improve function, they require prolonged treatment and vary in effectiveness from person to person. In summary, while these methods improve symptoms, they all have certain side effects and limitations.

[0005] The gut microbiome may play a significant role in the development and progression of Parkinson's disease. Parkinson's disease patients often experience gastrointestinal dysfunction, such as dysphagia, nausea, vomiting, and constipation. Inflammatory cytokines in the gut, such as IL-1β, can cross the blood-brain barrier and enter the brain, causing microglia in the brain to transition to an M1 activation state. This in turn mediates and triggers a neuroinflammatory response, further releasing large amounts 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. Furthermore, Parkinson's disease patients often experience a degree of gut microbial imbalance. Gut microbial imbalance has been observed in both Parkinson's disease patients and Parkinson's disease model mice, with increased abundance of harmful bacteria such as Escherichia coli and Shigella, and decreased abundance of beneficial bacteria such as Bifidobacterium and Akkermansia.

[0006] Probiotics not only regulate immunity but also reduce inflammation. They may interact with the brain through the "gut-brain axis," enhancing vagus nerve function and improving neuroinflammation, thereby positively impacting neurodegenerative diseases such as Parkinson's disease. Compared with traditional Parkinson's disease treatments, probiotic therapy has the advantages of fewer side effects and is less likely to cause drug resistance or dependence. As a non-invasive treatment, probiotics have a wide range of applications, can be used long-term, and are not easily restricted by the patient's physical condition. Therefore, it is very meaningful to develop more probiotic intervention strategies that can improve Parkinson's disease. Summary of the Invention

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

[0008] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

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

[0010] The present invention has developed a new probiotic compounding method and a new probiotic intervention strategy for improving Parkinson's disease, which is to combine Akkermansia muciniphila with Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve The BBr60 strain was compounded and found to have a potential interaction between the two, which can cooperate with each other to synergize the efficacy of improving Parkinson's disease. Specifically, it is manifested in: (1) synergistic improvement of movement disorders caused by Parkinson's disease; (2) synergistic reduction of the expression of related inflammatory factors in the substantia nigra, synergistic promotion of the production of anti-inflammatory factors to alleviate neuroinflammation; (3) synergistic inhibition of the activation of the inflammasome NLPR3 in the substantia nigra, thereby further inhibiting brain inflammation. At the same time, as probiotics, Akkermansia muciniphila Akk11 and Bifidobacterium breve BBr60 are highly safe and not prone to resistance.

[0011] Preferably, the Akkermansia muciniphila Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve The ratio of the number of viable bacteria of the BBr60 strain is 1:3-2:1, for example, 1:3, 2:5, 1:2, 2:3, 1:1, 3:2, 2:1, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described here one by one.

[0012] Based on the potential synergistic relationship between the two strains mentioned above, when they meet the above-mentioned specific ratio of live bacteria counts, they are more effective in improving movement disorders caused by Parkinson's disease, reducing the expression of related inflammatory factors in the substantia nigra, promoting the production of anti-inflammatory factors, and inhibiting the activation of inflammasome NLPR3 in the substantia nigra.

[0013] Preferably, the total number of viable bacteria in the probiotic preparation 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 this numerical range can be selected and will not be described here one by one.

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

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

[0016] 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.

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

[0018] 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.

[0019] Preferably, the probiotic preparation further contains excipients; the excipients include any one or a combination of at least two of fillers, binders, wetting agents, disintegrants, emulsifiers, cosolvents, solubilizers, osmotic pressure regulators, colorants, pH regulators, antioxidants, antibacterial agents or buffers.

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

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

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

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

[0024] Preferably, the Akkermansia muciniphila Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve The ratio of the number of viable bacteria of the BBr60 strain is 1:3-2:1, for example, 1:3, 2:5, 1:2, 2:3, 1:1, 3:2, 2:1, etc. Other specific point values ​​within this numerical range can be selected, and they will not be described here one by one.

[0025] Preferably, the total number of viable bacteria in the probiotic preparation 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), 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 this numerical range can be selected and will not be described here one by one.

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

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

[0028] The present invention has developed a new probiotic compounding method and a new probiotic intervention strategy for improving Parkinson's disease, which is to combine Akkermansia muciniphila with Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve The BBr60 strain was compounded and found to have a potential interaction between the two, which can cooperate with each other to synergize the efficacy of improving Parkinson's disease. Specifically, it is manifested in: (1) synergistic improvement of movement disorders caused by Parkinson's disease; (2) synergistic reduction of the expression of related inflammatory factors in the substantia nigra, synergistic promotion of the production of anti-inflammatory factors to alleviate neuroinflammation; (3) synergistic inhibition of the activation of the inflammasome NLPR3 in the substantia nigra, thereby further inhibiting brain inflammation. At the same time, as probiotics, Akkermansia muciniphila Akk11 and Bifidobacterium breve BBr60 are highly safe and not prone to resistance.

[0029] The Akk11 strain involved in the present invention is named Akkermansia muciniphila Akk11 Akkermansia muciniphila Akk11, the depository is China Center for Type Culture Collection, the deposit date is January 15, 2024, the deposit number is CCTCC NO: M2024119, and the address is: Wuhan University, Wuhan, China.

[0030] The BBr60 strain involved in the present invention is classified and named Bifidobacterium breve Bifidobacterium breve The depository is the General Microbiology Center of China Culture Collection Administration of Microorganisms. The deposit date is August 29, 2016. The deposit number is CGMCC No. 12915. The address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a statistical graph of the time it takes for mice in each group to fall from the pole 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 solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely to help understand the present invention and should not be regarded as specific limitations of the present invention.

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

[0047] The BBr60 strain involved in the following content is named Bifidobacterium breve Bifidobacterium breve The deposit date is August 29, 2016, and the deposit number is CGMCC No.12915.

[0048] The bacterial suspension preparation method involved in the following experiment is as follows: the desired strain is inoculated into MRS liquid medium, cultured at 37°C for 20 h for activation, and activated twice continuously to obtain an activated solution; the activated solution is inoculated into MRS liquid medium and cultured at 37°C to obtain a bacterial solution; the bacterial solution is centrifuged and filtered to obtain bacterial cells, and the bacterial cells are resuspended in PBS to obtain a bacterial suspension.

[0049] MRS medium: peptone 10 g / L, beef extract 10 g / L, glucose 20 g / L, sodium acetate 2 g / L, yeast powder 5 g / L, diammonium hydrogen citrate 2 g / L, K2PO4·3H2O 2.6 g / L, MgSO4·7H2O 0.1 g / L, MnSO4 0.05 g / L, Tween 80 1 mL / L, cysteine ​​hydrochloride 0.5 g / L.

[0050] Example

[0051] This example explores the improvement effect of probiotic preparations on Parkinson's disease mouse models:

[0052] (1) Experimental animals:

[0053] SPF male C57BL / 6J mice (5 weeks old, purchased from Jiangsu Jicui Pharmaceutical Kang Biotechnology Co., Ltd.) were first orally administered with 200 μL of 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 housed in a pathogen-free environment (12 h light / dark cycle) with room temperature (22 ± 1°C) and humidity (55 ± 5%). The animals had free access to food and water.

[0054] (2) Animal grouping:

[0055] Adaptive feeding was performed on days 1-7 of the experiment. After one week of adaptive feeding, the mice were randomly divided into 8 groups, with 8 mice in each group: untreated group (NC group), MPTP model group, Akk11 group (model mice were intervened with Akk11 bacterial solution), BBr60 group (model mice were intervened with BBr60 bacterial solution), composite group 1 (model mice were intervened with Akk11 bacterial solution and BBr60 bacterial solution, with a ratio of viable bacteria of 1:3), composite group 2 (model mice were intervened with Akk11 bacterial solution and BBr60 bacterial solution, with a ratio of viable bacteria of 2:1), composite control group (model mice were intervened with Akk11 bacterial solution and commercially available Bifidobacterium breve ATCC15700 bacterial solution, with a ratio of viable bacteria of 1:3), and probiotic control group (healthy mice were intervened with Akk11 bacterial solution and BBr60 bacterial solution, with a ratio of viable bacteria of 1:3).

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

[0057] Untreated group: From day 7 to day 35 of the experiment, mice were gavaged with biological saline every day;

[0058] MPTP model group: From day 14 to day 19 of the experiment, mice were intraperitoneally injected with MPTP (30 mg / kg) and probenecid (250 mg / kg) daily. From day 7 to day 35 of the experiment, mice were gavaged with biological saline daily.

[0059] Akk11 group: On days 14-19 of the experiment, mice were intraperitoneally injected with MPTP (30 mg / kg) and probenecid (250 mg / kg) daily. On days 7-35 of the experiment, Akk11 bacterial suspension (1×10 9 CFU / day / animal);

[0060] BBr60 group: From day 14 to day 19 of the experiment, mice were intraperitoneally injected with MPTP (30 mg / kg) and probenecid (250 mg / kg) every day. From day 7 to day 35 of the experiment, mice were gavaged with BBr60 suspension (1×10 9 CFU / day / animal);

[0061] Combined group 1: On days 14-19 of the experiment, mice were intraperitoneally injected with MPTP (30 mg / kg) and probenecid (250 mg / kg) daily. On days 7-35 of the experiment, mice were gavaged with a mixed suspension of Akk11 and BBr60 (total intervention dose 1×10 9 CFU / day / animal, the ratio of viable bacteria count is 1:3);

[0062] Combined group 2: On days 14-19 of the experiment, mice were intraperitoneally injected with MPTP (30 mg / kg) and probenecid (250 mg / kg) daily. On days 7-35 of the experiment, mice were gavaged with a mixed suspension of Akk11 and BBr60 (total intervention dose 1×10 9 CFU / day / animal, the ratio of viable bacteria count is 2:1);

[0063] Combined control group: From day 14 to day 19 of the experiment, mice were intraperitoneally injected with MPTP (30 mg / kg) and probenecid (250 mg / kg) daily. From day 7 to day 35 of the experiment, mice were gavaged with a mixed bacterial suspension of Akk11 and ATCC15700 (total intervention amount 1×10 9 CFU / day / animal, the ratio of viable bacteria count is 1:3);

[0064] Probiotic control group: From day 7 to day 35 of the experiment, mice were gavaged with Akk11 and BBr60 mixed bacterial suspension every day (total intervention amount 1×10 9 CFU / day / animal, the ratio of viable bacteria count is 1:3).

[0065] (4) Motor behavior test:

[0066] (4.1) Pole climbing test:

[0067] Prepare a vertical metal pole of about 50 cm and 1 cm in diameter. Two days before the test, all mice received adaptive training to minimize the impact of environmental changes on the experimental results. Specifically, the mice were placed in a rod cage to familiarize themselves with the environment, and then the mice were placed head up on a pole 15 cm from the bottom of the cage for 3 times, and then placed on poles 30 cm and 50 cm above the cage floor for 3 training sessions. On the day of the test, each mouse was placed head up on the top of the pole, and their descent was observed. The time until the two front paws touched the bottom ground was recorded. The test was repeated 3 times with an interval of 5 minutes between each trial. If the mouse fell or jumped off the pole within 60 seconds, the test was repeated, the average time was taken, and statistical analysis was performed. The results are as follows. Figure 1 shown.

[0068] (4.2) Rotating rod test:

[0069] The RT-01 mouse rotarod, which includes an automatic timer and a fall sensor, was used during both training and formal testing. Prior to formal testing, mice received three training sessions, each lasting 180 seconds per day for three days at three incremental speeds (10, 20, and 30 rpm). During the formal experiment, the rotarod was set to 30 rpm, and mice were placed on the rotarod. The latency to fall was recorded and averaged. Figure 2 shown.

[0070] (4.3) Open field test:

[0071] Two days before the test, the mice were trained to adapt to the environment to reduce the stress brought by the new environment in the formal experiment. The open field consists of a square arena with an area of ​​50 cm × 50 cm. The arena has four walls, each 45 cm high. During the formal test, the laboratory maintains a quiet and properly lit environment, and human intervention is minimized. The mouse is gently placed in the center of the open field, and its behavior is recorded for 5 minutes. The total distance and number of entries of the mouse in the center are recorded. Figure 3 and Figure 4 shown.

[0072] Depend on Figures 1-4 The results showed that compared with the NC group, the time for the model group mice to descend the rod was significantly prolonged, the time for falling on the rotating rod was significantly shortened, and the performance was poor in terms of total moving distance and the number of times entering the open area; after intervention with probiotics in each group, the time for descending the rod was shortened to varying degrees, the time for falling on the rotating rod was prolonged to varying degrees, and the total moving distance and the number of times entering the open area of ​​the mice increased to varying degrees. The improvement effects of the compound 1 group and the compound 2 group were more obvious than those of the Akk11 group and the BBr60 group, indicating that the Akk11 strain and the BBr60 strain 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 killed and the mouse tissues were collected for analysis. The total RNA in the substantia nigra tissue of the mouse brain was extracted using a kit. The kit was then used for reverse transcription and finally for qRT-PCR analysis. The expression of related genes was normalized with the level of β-actin. The results are shown in Figure 2. Figure 5-10 shown.

[0075] Depend on Figure 5-10 The results showed that 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 the intervention of probiotics in each group, the levels of pro-inflammatory factors decreased to varying degrees, and the levels of anti-inflammatory factors increased to varying degrees, and the improvement effects of the compound 1 group and the compound 2 group were more obvious than those of the Akk11 group and the BBr60 group, indicating that the Akk11 strain and the BBr60 strain cooperate and promote each other in alleviating the inflammatory response of Parkinson's mice.

[0076] (6) Inflammasome protein immunoblotting assay:

[0077] The substantia nigra tissue of each group of mice was 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 for 2 hours at room temperature. Then the membrane was incubated overnight with the following primary antibodies: β-actin (1:3000), NLRP3 (1:1000), caspase-1 (1:1000), IL-1β (1:1000), and incubated overnight at 4°C. The membrane was then incubated with secondary antibodies of anti-mouse IRDye®680RD conjugated antibody and anti-rabbit IRDye®800CW conjugated antibody. The immunoblot was scanned using a dual-color infrared laser imaging scanner and analyzed using software. The statistical results of the relative expression of each group of proteins are shown in Figure 2. Figure 11-13 shown.

[0078] NLRP3 inflammasome plays an important role in the pathogenesis of Parkinson's disease, mainly accelerating the death of dopaminergic neurons through microglial activation, release of proinflammatory factors, mitochondrial damage and cell pyroptosis. Figure 7 The results showed that compared with the NC group, the inflammation-related protein NLRP3 level, Caspase-1 / pro-Caspase-1 ratio level and IL-1β / pro-IL-1β ratio level of the model group mice were significantly increased; probiotic intervention in each group reversed the expression trend of these proteins, and the reversal effect of compound 1 and compound 2 groups was more obvious than that of Akk11 group and BBr60 group, indicating that Akk11 strain and 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 present invention is illustrated by the above-described embodiments, but the present invention is not limited to the above-described embodiments. This does not mean that the present invention must rely on the above-described embodiments in order to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent replacements for raw materials in the present invention, additions of auxiliary ingredients, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

[0080] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0081] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A probiotic preparation for improving Parkinson's disease, characterized in that: The strains in the probiotic preparation include Akkermansia muciniphila with a deposit number of CCTCC NO: M2024119 Akkermansia muciniphila Akk11 strain and Bifidobacterium breve with the deposit number of CGMCC No.12915 Bifidobacterium breve BBr60 strain.

2. The probiotic preparation for improving Parkinson's disease according to claim 1, characterized in that Akkermansia muciniphila Akkermansia muciniphila Akk11 strain and Bifidobacterium breve Bifidobacterium breve The ratio of viable bacteria count of BBr60 strain is 1:3-2:

1.

3. 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.

4. 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.

5. 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.

6. 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.

7. 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.

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

9. 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.

10. Use of the probiotic preparation for improving Parkinson's disease according to any one of claims 1 to 9 in the preparation of an NLRP3 inflammasome activation inhibitor.

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