Strains, compositions, uses and products for the prevention and treatment of parkinson's disease

Abstract

The application discloses strains for preventing and treating Parkinson's disease and compositions, applications and products thereof, and relates to the technical field of microorganisms.The application provides three strains of human milk origin, namely Lactobacillus helveticus LV-20, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15.The application also provides a composition capable of preventing and treating Parkinson's disease.The three strains can preferably prevent and treat Parkinson's disease.The composition can significantly improve motor dysfunction (such as static tremor, muscle rigidity, bradykinesia, posture balance and the like), reduce neuron damage, significantly reduce alpha-syn deposition, reverse sensory function degradation and cognitive impairment, relieve sleep disorders, inhibit neural inflammation and oxidative stress, reduce and repair intestinal barrier damage, relieve gastrointestinal dysfunction, improve constipation and regulate intestinal flora composition, and has a wide application prospect as a probiotic or synbiotic product for treating and preventing Parkinson's disease.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbiology, in particular to a strain and its composition, application and product for preventing and treating Parkinson's disease. BACKGROUND

[0002] Parkinson's Disease (PD) is a common neurodegenerative disease, mainly characterized by the progressive loss of dopaminergic neurons in the substantia nigra and the abnormal aggregation of alpha-synuclein in the midbrain. Currently, the clinical treatment of PD mainly relies on drugs and surgery (such as deep brain stimulation), drugs including levodopa, dopamine receptor agonists, MAO-B inhibitors and COMT inhibitors. Levodopa, a dopamine precursor, can effectively improve motor symptoms, but long-term use can lead to "wearing-off" and dyskinesia. Dopamine receptor agonists (such as pramipexole, ropinirole) can directly stimulate dopamine receptors, but may cause impulse control disorders (such as pathological gambling, compulsive shopping). MAO-B inhibitors (such as selegiline) and COMT inhibitors (such as entacapone) can prolong the action of dopamine, but have limited efficacy and may cause liver toxicity. Drugs and surgery can alleviate symptoms, but cannot stop disease progression, and long-term use may lead to motor complications (such as dyskinesia, wearing-off) and mental side effects (such as hallucinations, impulse control disorders).

[0003] Recent studies have shown that gut dysbiosis is closely related to the occurrence and development of PD, i.e. the "gut-brain axis" hypothesis. PD patients often have constipation, intestinal inflammation and other gastrointestinal symptoms, and their intestinal flora composition is significantly different from that of healthy people, such as decreased Prevotella and increased Enterobacteriaceae. These changes in flora may exacerbate neurodegenerative lesions by promoting the misfolding of alpha-synuclein, activating neuroinflammation (such as microglial activation) and affecting the production of short-chain fatty acids (SCFAs).

[0004] Intervention strategies targeting gut microbiota (such as probiotics, prebiotics) have become a research hotspot, but the existing solutions have the following problems: (1) Single strain: Most studies use a single strain (such as lactobacillus, bifidobacterium), which is difficult to comprehensively regulate the complex intestinal microecology. (2) Lack of specificity: Most existing probiotic products are aimed at gastrointestinal health, rather than specifically regulating PD-related flora-neuroinflammatory pathways (such as TNF-alpha, IL-6). (3) Single effect: Probiotics can only improve individual symptoms when treating PD patients, and it is difficult to produce sustained and effective changes in most symptoms. For example, a clinical trial (NCT03575195) used a complex probiotic (containing L. acidophilus, B. bifidum, etc.) to treat PD patients, which improved constipation symptoms, but had no significant effect on motor function. (4) Lack of effective combination: Probiotic combinations or prebiotic combinations are more commonly used to prevent and treat symptoms in PD patients, but there are few reports of their combination for preventing and treating symptoms in PD patients. SUMMARY

[0005] The purpose of the present application is to provide strains and their compositions, applications and products for preventing and treating Parkinson's disease.

[0006] To achieve the above-mentioned purposes of the application, the technical solutions of the present application are as follows:

[0007] In one aspect, the present application provides a strain of Lactobacillus helveticus LV-20, the preservation number of which is CGMCC No: 34563.

[0008] Specifically, the colony characteristics of the Lactobacillus helveticus LV-20 on the MRS medium are white, near-circular, wet surface, opaque, and irregular edge.

[0009] In another aspect, the present application provides a strain of Lactobacillus plantarum LPP8, the preservation number of which is CGMCC No: 34565.

[0010] Specifically, the colony characteristics of the Lactobacillus plantarum LPP8 on the MRS medium are white or light yellow, circular, wet surface, opaque, and regular edge.

[0011] In another aspect, the present application provides a strain of Lactobacillus fermentum PL-15, the preservation number of which is CGMCC No: 34564.

[0012] Specifically, the colony characteristics of the Lactobacillus fermentum PL-15 on the MRS medium are white, circular, wet surface, opaque, and regular edge.

[0013] In another aspect, the present invention provides a product for the prevention or treatment of Parkinson's disease, the product comprising one or more of the above-mentioned Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8, and Lactobacillus fermentum PL-15.

[0014] Furthermore, the product can improve motor dysfunction.

[0015] Furthermore, the viable count of *Lactobacillus helveticus* LV-20, *Lactobacillus plantarum* LPP8, or *Lactobacillus fermentum* PL-15 in the product is 1 × 10⁻⁶. 5 -10 12 CFU / g;

[0016] Furthermore, the viable count of *Lactobacillus helveticus* LV-20, *Lactobacillus plantarum* LPP8, or *Lactobacillus fermentum* PL-15 in the product is 1 × 10⁻⁶. 7 -10 10 CFU / g;

[0017] Furthermore, the viable count of *Lactobacillus helveticus* LV-20, *Lactobacillus plantarum* LPP8, or *Lactobacillus fermentum* PL-15 in the product is 9 × 10⁻⁶. 9 CFU / g.

[0018] Specifically, the product may also include cultures of Lactobacillus helveticus LV-20, Lactobacillus plantarum LPP8, or Lactobacillus fermentum PL-15.

[0019] Furthermore, the culture refers to the entire culture medium containing the strain, strain extracts, its metabolites, additional nutrients, etc., obtained by culturing the strain (Lactobacillus helveticus LV-20, Lactobacillus plantarum LPP8, or Lactobacillus fermentum PL-15) for a period of time in a medium that can supply nutrients so that the strain (Lactobacillus helveticus LV-20, Lactobacillus plantarum LPP8, or Lactobacillus fermentum PL-15) can grow and survive, but also includes the culture medium obtained by eliminating the strain after culturing the strain.

[0020] Culture medium can refer to the upper liquid itself, excluding the portion that settles to the bottom after being left to stand for a period of time; or it can refer to the culture medium obtained by removing bacterial cells through filtration; or it can refer to the upper liquid itself obtained by centrifuging the culture medium to remove the lower sediment. Culture medium can be used as a concentrate by concentrating it using common methods.

[0021] Culture medium or a concentration of culture medium can be provided as a dried product by drying using common methods.

[0022] The culture described in the present application can include a culture solution of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 or Lactobacillus fermentum PL-15, a concentrate of the culture solution, a dry product of the culture solution or the concentrate.

[0023] Specifically, the product is a pharmaceutical composition.

[0024] Further, the pharmaceutical composition can further include a pharmaceutically acceptable excipient.

[0025] Still further, the pharmaceutically acceptable excipient includes, but is not limited to, an excipient, a disintegrant, an antioxidant, a pH adjuster, a diluent, an osmotic pressure adjuster, an emulsifier, a binder, a film-forming agent, a wetting agent.

[0026] Specifically, the dosage form of the pharmaceutical composition includes, but is not limited to, a gastrointestinal administration dosage form or a non-gastrointestinal administration dosage form according to the administration method.

[0027] Further, the gastrointestinal administration dosage form includes, but is not limited to, a powder, a tablet, a granule, a capsule, a solution, an emulsion, a suspension, an oil.

[0028] Further, the non-gastrointestinal administration dosage form includes, but is not limited to, an injection administration dosage form, a respiratory administration dosage form, a skin administration dosage form, a mucosa administration dosage form, a cavity administration dosage form.

[0029] Still further, the injection administration dosage form includes, but is not limited to, an intravenous injection, an intramuscular injection, a subcutaneous injection, an intradermal injection, an intracavity injection.

[0030] Still further, the respiratory administration dosage form includes, but is not limited to, a spray, an aerosol, a powder spray.

[0031] Still further, the skin administration dosage form includes, but is not limited to, an external solution, a lotion, a liniment, an ointment, a plaster, a paste, a patch.

[0032] Still further, the mucosa administration dosage form includes, but is not limited to, an eye drop, a nose drop, an eye ointment, a gargle, a sublingual tablet, a sticking tablet and a film.

[0033] Still further, the cavity administration dosage form includes, but is not limited to, a suppository, an aerosol, an effervescent tablet, a drop and a drop pill.

[0034] In another aspect, the present application provides a use of the above-mentioned Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 or Lactobacillus fermentum PL-15 in the preparation of a product for preventing or treating Parkinson's disease.

[0035] Further, the product can improve motor dysfunction.

[0036] In particular, the product is a pharmaceutical composition.

[0037] Further, the pharmaceutical composition can further comprise a pharmaceutically acceptable excipient.

[0038] Still further, the pharmaceutically acceptable excipient includes but is not limited to an excipient, a disintegrant, an antioxidant, a pH regulator, a diluent, an osmotic pressure regulator, an emulsifier, a binder, a film-forming agent, a wetting agent.

[0039] In particular, the dosage form of the drug includes but is not limited to a gastrointestinal administration dosage form or a non-gastrointestinal administration dosage form according to the administration mode.

[0040] Further, the gastrointestinal administration dosage form includes but is not limited to a powder, a tablet, a granule, a capsule, a solution, an emulsion, a suspension, an oil.

[0041] Further, the non-gastrointestinal administration dosage form includes but is not limited to an injection administration dosage form, a respiratory administration dosage form, a skin administration dosage form, a mucosa administration dosage form, a cavity administration dosage form.

[0042] Still further, the injection administration dosage form includes but is not limited to an intravenous injection, an intramuscular injection, a subcutaneous injection, an intradermal injection, an intracavity injection.

[0043] Still further, the respiratory administration dosage form includes but is not limited to a spray, an aerosol, a powder spray.

[0044] Still further, the skin administration dosage form includes but is not limited to a topical solution, a lotion, a liniment, an ointment, a plaster, a paste, a patch.

[0045] Still further, the mucosa administration dosage form includes but is not limited to an eye drop, a nose drop, an eye ointment, a gargle, a sublingual tablet, a sticking tablet and a film patch.

[0046] Still further, the cavity administration dosage form includes but is not limited to a suppository, an aerosol, an effervescent tablet, a drop and a drop pill.

[0047] In another aspect, the present application provides a composition comprising a composite bacteria powder, wherein the composite bacteria powder comprises the above-mentioned Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15.

[0048] In particular, the viable bacterial count of the Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the composite bacteria powder is 1×10 5 -10 12 CFU / g.

[0049] Further, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is (0.5-10):(0.5-10):1;

[0050] Further, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is (0.5-5):(0.5-5):1;

[0051] Further, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is (0.5-2):(0.5-2):1;

[0052] Specifically, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is 1:1:1, 2:1:1, 1:2:1 or 0.5:0.5:1.

[0053] According to some embodiments of the present application, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is 3x10 9 CFU: 3x10 9 CFU: 3x10 9 CFU.

[0054] According to some embodiments of the present application, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is 4.5x10 9 CFU: 2.25x10 9 CFU: 2.25x10 9 CFU.

[0055] According to some embodiments of the present application, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is 2.25x10 9 CFU: 4.5x10 9 CFU: 2.25x10 9 CFU.

[0056] According to some embodiments of the present application, the viable cell number ratio of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the complex bacterial powder is 2.25x10 9 CFU: 2.25x10 9CFU: 4.5 x 10 9 CFU.

[0057] Specifically, the composition further includes an adjuvant.

[0058] Further, the adjuvant includes, but is not limited to, fructo-oligosaccharide, stachyose, galacto-oligosaccharide, resistant dextrin, mannan, pentosan, beta-glucan, arabinoxylan.

[0059] Still further, the adjuvant includes fructo-oligosaccharide, stachyose, galacto-oligosaccharide, and resistant dextrin.

[0060] Specifically, the weight ratio of the complex bacterial powder to fructo-oligosaccharide, stachyose, galacto-oligosaccharide, and resistant dextrin is (5-10):(1-3):(1-3):(1-3):1.

[0061] Further, the weight ratio of the complex bacterial powder to fructo-oligosaccharide, stachyose, galacto-oligosaccharide, and resistant dextrin is (5-8):(1-2):(1-2):(1-2):1.

[0062] Still further, the weight ratio of the complex bacterial powder to fructo-oligosaccharide, stachyose, galacto-oligosaccharide, and resistant dextrin is 6:1:1:1:1.

[0063] Specifically, the composition can further include a culture of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8, or Lactobacillus fermentum PL-15.

[0064] Further, the culture means the entire culture medium containing the strain, a strain extract, metabolites thereof, additional nutritional ingredients, etc., obtained by culturing the strain (Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8, or Lactobacillus fermentum PL-15) in a culture medium capable of supplying nutritional ingredients so that the strain can grow and survive for a certain period of time, but also includes the culture solution obtained by eliminating the strain after culturing the strain.

[0065] The culture solution can mean the supernatant itself collected except for the portion that sinks to the bottom layer by being left for a certain period of time, or means the culture solution obtained by removing the bacterial bodies by filtration, or means the upper liquid itself obtained by centrifuging the culture solution to remove the lower precipitate. The culture solution can be used as a concentrate by being concentrated in a common method.

[0066] The culture solution or the concentrate of the culture solution can be provided as a dry product by being dried in a common method.

[0067] The culture described in the present application can include a culture solution of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 or Lactobacillus fermentum PL-15, a concentrate of the culture solution, a dry product of the culture solution or the concentrate.

[0068] Specifically, the composition is a synbiotic.

[0069] According to some embodiments of the present application, the preparation method of the synbiotic is mixing the compound bacterial powder with fructo-oligosaccharide, stachyose, galacto-oligosaccharide and resistant dextrin to obtain the synbiotic.

[0070] According to some embodiments of the present application, the preparation method of the bacterial powder is: inoculating Lactobacillus helveticus LV-20, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 activated respectively into MRS liquid medium with an inoculation amount of 1%, culturing at 37°C for 24 hours, centrifuging to obtain bacterial bodies, adding a freeze-drying protective agent to resuspend the bacterial bodies according to a weight ratio of 1:1 of the bacterial body wet weight to the freeze-drying protective agent weight, and placing the bacterial bodies into a freeze-drying machine to obtain the bacterial powder by freeze-drying.

[0071] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for treating or preventing Parkinson's disease.

[0072] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for improving motor dysfunction.

[0073] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for reducing pathological features of brain tissues.

[0074] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for inhibiting neuroinflammation.

[0075] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for resisting oxidative stress.

[0076] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for repairing intestinal barrier.

[0077] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for improving constipation.

[0078] In another aspect, the present application provides use of the above-mentioned composition in preparation of a product for regulating composition of intestinal flora.

[0079] Specifically, the product is a drug.

[0080] Further, the drug can further include a pharmaceutically acceptable excipient.

[0081] Further, the pharmaceutically acceptable adjuvant includes, but is not limited to, excipients, disintegrants, antioxidants, pH regulators, diluents, osmotic pressure regulators, emulsifiers, binders, film formers, wetting agents.

[0082] Specifically, the dosage form of the drug includes, but is not limited to, a gastrointestinal administration dosage form or a non-gastrointestinal administration dosage form according to the administration mode.

[0083] Further, the gastrointestinal administration dosage form includes, but is not limited to, a powder, a tablet, a granule, a capsule, a solution, an emulsion, a suspension, an oil.

[0084] Further, the non-gastrointestinal administration dosage form includes, but is not limited to, an injection administration dosage form, a respiratory administration dosage form, a skin administration dosage form, a mucosa administration dosage form, a cavity administration dosage form.

[0085] Further, the injection administration dosage form includes, but is not limited to, an intravenous injection, an intramuscular injection, a subcutaneous injection, an intradermal injection, an intracavity injection.

[0086] Further, the respiratory administration dosage form includes, but is not limited to, a spray, an aerosol, a powder spray.

[0087] Further, the skin administration dosage form includes, but is not limited to, an external solution, a lotion, a liniment, an ointment, a plaster, a paste, a patch.

[0088] Further, the mucosa administration dosage form includes, but is not limited to, an eye drop, a nose drop, an eye ointment, a gargle, a sublingual tablet, a sticking tablet and a film.

[0089] Further, the cavity administration dosage form includes, but is not limited to, a suppository, an aerosol, an effervescent tablet, a drop and a drop pill.

[0090] The beneficial effects of the present application are:

[0091] The present application provides Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15, and verifies the preventive effect of the three strains on Parkinson's disease, and the three strains have improving effects on behavior and cognitive impairment of Parkinson's disease, and can better prevent Parkinson's disease.

[0092] The present application provides a synbiotic for preventing and treating Parkinson's disease, which comprises Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8, Lactobacillus fermentum PL-15 and adjuvants, and the synbiotic has the following effects in preventing or treating Parkinson's disease:

[0093] (1) significantly improving motor dysfunction

[0094] (2) reducing brain tissue pathological characteristics

[0095] Synbiotic can restore tyrosine hydroxylase expression in substantia nigra and striatum, reduce neuronal damage, and significantly reduce α-syn deposition;

[0096] (3) Inhibition of neuroinflammation and oxidative stress

[0097] Synbiotic can inhibit inflammasome pathway: down-regulate NLRP3, caspase-1, IL-1β, IL-18 expression, reduce pro-inflammatory factors (TNF-α, IL-6), and increase anti-inflammatory factors (IL-10);

[0098] Synbiotic has antioxidant effect: activation of Nrf2 / ARE pathway, increase of SOD and GSH levels, and decrease of MDA level;

[0099] (4) Reduction and repair of intestinal barrier damage

[0100] Synbiotic can repair intestinal barrier: restore colon length, significantly increase mucin and tight junction protein (ZO-1, Occludin) expression; reduce serum LPS level, reduce leaky gut syndrome, and reduce LPS content in blood;

[0101] (5) Alleviation of gastrointestinal dysfunction: increase fecal output and improve fecal water content;

[0102] (6) Regulation of intestinal flora composition

[0103] Synbiotic can restore flora richness, reduce harmful bacteria (such as Enterobacteriaceae), and increase beneficial bacteria (such as Akkermansia, Bifidobacterium); significantly optimize metabolism and immune-related pathways. BRIEF DESCRIPTION OF DRAWINGS

[0104] Figure 1 For the behavioral test results of Example 2, A-B in the figure are pole climbing experiments, ns>0.05, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

[0105] Figure 2 For the behavioral test results of Example 2, A is the rotarod test, and B is the open field test, ns: P>0.05, *P<0.05, **P<0.01, ****P<0.0001.

[0106] Figure 3 For the behavioral test results in Example 3, A-B in the figure are the pole climbing test results, C is the rotarod test result, D1, D2, D3, E, and F are the open field test results; *P<0.05, **P<0.01, ***P<0.001.

[0107] Figure 4 Effect of synbiotic on dopamine production in PD mice, A-D are TH expression in SN and striatum; E is WB detection of TH protein expression in SN; *P<0.05, **P<0.01, ***P<0.001; A scale is 50um, B scale is 250um.

[0108] Figure 5 Effect of synbiotic on a-syn aggregation in PD mice, A and C are a-syn expression in SN; B is a-syn expression in striatum; D is the proportion of a-syn expression in striatum; E is WB detection of a-syn protein expression in SN; *P<0.05, **P<0.01, ****P<0.0001; A scale is 100um, B scale is 250um.

[0109] Figure 6 Effect of synbiotic on neuroinflammation in PD mice, A is iba-1 expression in SN; B is GFAP expression in SN; C is Nissl body expression in SN; D-F are quantification figures, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001; Scale in A-C is 50um.

[0110] Figure 7 Effect of synbiotic on inflammation and pyroptosis pathways in PD mice, the figure is the expression of NLRP3 and other pyroptosis-related proteins in SN.

[0111] Figure 8 Effect of synbiotic on inflammation and pyroptosis pathways in PD mice, A-I are the expression of NLRP3 and other pyroptosis-related proteins in SN, *P<0.05, **P<0.01, ***P<0.001.

[0112] Figure 9 Effect of synbiotic on inflammation and pyroptosis pathways in PD mice, A-F are the expression of IL-6, IL-10, TNF-a, IL-1β, IL-18, NLRP3, caspase1, GSDMD mRNA in SN; P is the content of LPS in SN; ns: P>0.05, *P<0.05, **P<0.01, ***P<0.001.

[0113] Figure 10 Effect of synbiotic on oxidative stress-related pathway proteins in PD mice; the figure is the expression of Nrf2 pathway-related proteins in SN.

[0114] Figure 11Effects of Synbiotic on the oxidative stress related pathway proteins in PD mice; A-E are the expression of Nrf2 pathway related proteins in the substantia nigra; F-I are the contents of SOD, GSH, MDA and NQO1 in serum; ns: P>0.05, *P<0.05, **P<0.01, ***P<0.001.

[0115] Figure 12 Effects of Synbiotic on the colon length, structure and mucin secretion in PD mice; A and D are the colon length; B is the HE staining of colon; C and E are the Alcian blue staining of colon; *P<0.05, **P<0.01, ***P<0.001; the scale bar in B and C is 50 um.

[0116] Figure 13 Effects of Synbiotic on the expression of tight junction proteins in the colon of PD mice; A-D are the immunofluorescence staining of ZO-1 and occludin in the colon; E is the mRNA expression of ZO-1 and occludin in the colon; F is the content of LPS in serum; *P<0.05, **P<0.01, ***P<0.001.

[0117] Figure 14 Fecal detection results; A is the fecal output; B is the water content of feces; C is the fecal particle morphology; *P<0.05, ***P<0.001.

[0118] Figure 15 Effects of Synbiotic on the intestinal flora composition of PD mice; A is the beta diversity; B is the relative abundance heatmap at the species level.

[0119] Figure 16 Effects of Synbiotic on the intestinal flora composition of PD mice; A-D are the violin plots of Bacteroides, Duncaniella, Muribaculaceae-bacterium, Muribaculaceae-bacterium-Isolate-013-NCI distribution in each group; *P<0.05, **P<0.01, ***P<0.001.

[0120] Figure 17 Behavioral test results of Example 4; A-B are the rotarod test, C is the open field test; *P<0.05, **P<0.01.

[0121] Figure 18 Effects of Synbiotic treatment on the pathological changes of PD mice; A is the expression of TH in the substantia nigra; B is the result of Nissl staining in the substantia nigra; *P<0.05, **P<0.01.

[0122] Figure 19 Synbiotic can inhibit NLRP3 inflammasome-mediated inflammatory response in PD mice; A-H in the figure are IL-6, IL-10, TNF-a, IL-1β, IL-18, NLRP3, caspase1, GSDMD mRNA expression in the substantia nigra; ns: P>0.05, *P<0.05, **P<0.01, ***P<0.001.

[0123] Figure 20 Effect of synbiotic treatment on intestinal barrier integrity; A is the length of the colon; B is the number of goblet cells; *P<0.05, ***P<0.001.

[0124] Figure 21 Effect of synbiotic treatment on intestinal tight junction proteins; A-B are the results of occludin and ZO-1 immunofluorescence staining quantification in colon; **P<0.01, ***P<0.001.

[0125] DEPOSIT DESCRIPTION

[0126] Deposit number: CGMCC No. 34563;

[0127] Classification name: Lactobacillus helveticus Lactobacillus helveticus ;

[0128] Date of deposit: May 19, 2025;

[0129] Deposit unit: China General Microbiological Culture Collection Center;

[0130] Abbreviation of deposit unit: CGMCC;

[0131] Address of deposit: No. 3, Beichen West Road, Chaoyang District, Beijing.

[0132] DEPOSIT DESCRIPTION

[0133] Deposit number: CGMCC No. 34564;

[0134] Classification name: Lactobacillus fermentum Lactobacillus fermentum ;

[0135] Date of deposit: May 19, 2025;

[0136] Deposit unit: China General Microbiological Culture Collection Center;

[0137] Abbreviation of deposit unit: CGMCC;

[0138] Address of deposit: No. 3, Beichen West Road, Chaoyang District, Beijing.

[0139] Deposit Description

[0140] Deposit Number: CGMCC No. 34565;

[0141] Classification Name: Lactiplantibacillus plantarum Lactiplantibacillus plantarum ;

[0142] Date of Deposit: May 19, 2025;

[0143] Deposit Unit: China General Microbiological Culture Collection Center;

[0144] Abbreviation of Deposit Unit: CGMCC;

[0145] Address of Deposit: No. 3, Beichen West Road, Beijing City, China. DETAILED DESCRIPTION

[0146] In order to make the technical means, creative features, purposes and effects realized by the present application easy to understand, the following specific embodiments are further illustrated, but the following embodiments are only preferred embodiments of the present application, not all. Based on the embodiments in the embodiments, other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application. In the following embodiments, if not otherwise specified, the operation method used is a conventional operation method, the equipment used is a conventional equipment, and the equipment and materials used in each embodiment are the same.

[0147] Example 1 Preparation of synbiotic

[0148] The activated Lactobacillus helveticus LV-20, Lactiplantibacillus plantarum LPP8 and Lactobacillus fermentum PL-15 were prepared into bacterial powder according to the following steps: 1% inoculation amount was inoculated into sterile MRS liquid medium, and cultured at 37°C for 24h, and then centrifuged to obtain bacterial bodies, and the bacterial bodies were resuspended with freeze-drying protectant (10% skim milk, 5% trehalose) according to the weight ratio of 1:1, and then placed in a freeze-drying machine for freeze-drying. Finally, the pour plate method was used for counting. The viable bacterial count of the three bacterial powders in the synbiotic was prepared in a weight ratio of 1:1:1, and the mixed bacterial powder was mixed with fructooligosaccharide, stachyose, galactooligosaccharide and resistant dextrin in a weight ratio of 6:1:1:1:1.

[0149] Example 2

[0150] Male C57BL / 6J mice (6-8 weeks old) were randomly divided into groups of 8 after one week of acclimation, including control group (NC), rotenone model group (PD), Lactobacillus helveticus LV-20 group, Lactobacillus plantarum LPP8 group, Lactobacillus fermentum PL-15 group, combination 1 (bacterial number LV-20:LPP8:PL-15 = 1:1:1), combination 2 (bacterial number LV-20:LPP8:PL-15 = 10:1:1), combination 3 (bacterial number LV-20:LPP8:PL-15 = 1:10:1), combination 4 (bacterial number LV-20:LPP8:PL-15 = 1:1:10), combination 5 (bacterial number LV-20:LPP8:PL-15 = 1:1:2), and combination 6 (bacterial number LV-20:LPP8:PL-15 = 20:1:1). At the beginning of the experiment, the probiotic groups were gavaged with 0.2 mL of bacterial solution (containing 9 x 10^9 CFU / d) in the morning, and the PD and NC groups were gavaged with the same volume of sterile PBS (0.2 mL). In the afternoon, the PD and SYN groups were gavaged with 30 mg / kg / d rotenone, and the NC group was gavaged with the same volume of rotenone solvent (4% chloroform and 0.5% carboxymethyl cellulose CMC). After 4 weeks, behavioral tests were performed.

[0151] From Figures 1-2 The results show that the three strains and four combinations have effects on the behavior and cognitive impairment of PD mice. Among the single strains, Lactobacillus helveticus LV-20 is more effective than Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15. Combinations 1, 2, 3, and 5 are effective, with combinations 1, 2, and 5 being more effective than combination 3, and combination 1 being the most effective. Combinations 4 and 6 have no effect.

[0152] Example 3 Synbiotic for preventing PD

[0153] 1. Male C57BL / 6J mice (6-8 weeks old) were randomly divided into groups of 15 after one week of acclimation, including control group (NC), rotenone model group (PD), and synbiotic intervention group (SYN). At the beginning of the experiment, the SYN group was gavaged with 0.2 mL of synbiotic (containing 9 x 10^9 CFU / d) in the morning, and the PD and NC groups were gavaged with the same volume of sterile PBS (0.2 mL). In the afternoon, the PD and SYN groups were gavaged with 30 mg / kg / d rotenone, and the NC group was gavaged with the same volume of rotenone solvent (4% chloroform and 0.5% carboxymethyl cellulose CMC). After 4 weeks, behavioral tests were performed, and the experiment in mice was completed after the tests. Serum, substantia nigra and striatal brain tissue, colon, and fecal particles were collected.

[0154] 2. Behavioral tests

[0155] Rope climbing test, a 50 cm long, 3 cm diameter wooden rod and a fixed wooden ball at the top of the rod were placed in the mouse cage. The head turning time of each mouse to lower the rod and the time spent by the mouse from the top to the bottom with one hind leg were recorded. The mice had been trained to descend from the top to the cage before the test. Three tests were conducted with an interval of 1 hour.

[0156] Rotarod test, the mice were placed on a rotating rod, and then the rod rotated at a speed of 30 rpm for 120 seconds. Subsequently, the rotating rod automatically recorded the time when each mouse fell off the rod and the number of times the mouse fell off. Each mouse received 3 tests with an interval of 1h.

[0157] Open field test, the animals were adapted to the stable feeding conditions for 1-3 days before the experiment. During the experiment, the animals were first placed in the open field for 5-10 minutes, then gently placed in the fixed corner of the open field, and the recording was started. The total distance, central area residence time and number of times, number of times of standing, and number of times of grooming behavior were recorded.

[0158] The longer the time for the mouse to climb from the top of the rod to the bottom of the rod in the rope climbing test, and the more times the mouse turned its head, the worse the mouse's motor coordination and balance function. The longer the mouse stays on the rotating rod in the rotarod test, the fewer the number of times the mouse falls off, the better the mouse's motor coordination and balance function. In the open field test, mice with depression, anxiety and high tension are afraid to explore the central open field area. As shown in Figure 3 , from Figure 3 A- Figure 3 F, it can be seen that compared with the PD group of mice, the behavior and cognitive impairment of the SYN intervention group were significantly improved, and the rope climbing, rotarod and open field tests were significantly better than the PD group.

[0159] 3. Brain tissue pathological detection

[0160] After the fresh brain tissue of the mice was soaked in paraformaldehyde for 48 hours, the sample was placed in a fully automatic dehydration machine for dehydration treatment. After the dehydration process was completed, the fully dehydrated tissue was embedded. After embedding, slicing (4 μm), deparaffinization, staining, microscopic observation and photographing were performed.

[0161] Tyrosine hydroxylase (TH) is a key enzyme in the synthesis of dopamine. In the process of Parkinson's disease, the decrease of dopamine production will also reduce the key enzyme TH. After antigen repair, endogenous peroxidase blocking, blocking (5% BSA solution, incubated at room temperature for 1h), the addition of primary antibody was incubated overnight at 4°C in a chromatography cabinet, washed with PBS, added biotin-labeled secondary antibody, incubated in the dark for 30min-1h, washed with PBS, added DAB for color development, and observed under a microscope and recorded photographs. The results of immunohistochemical staining showed that Figure 4 A- Figure 4D) Compared with the NC group, the PD group showed a significant decrease in TH in both the SN and the striatum (P < 0.01), while the SYN group after vitamin intervention showed a significant increase compared with the PD group (P < 0.05). The tissue protein sample was subjected to SDS-PAGE gel electrophoresis, electric transfer, and skimmed milk blocking. The first antibody was incubated at 4°C overnight, washed, and the second antibody was incubated at room temperature for 1 h. After washing, the hypersensitive ECL luminescent liquid was used for development, and the photographs were taken, saved, and analyzed. Figure 4 E) Western blotting experiments also demonstrated a significant decrease in TH in brain tissue proteins, showing consistent results.

[0162] An important pathological feature of PD development is the abnormal aggregation of alpha-synuclein. Immunohistochemical staining results showed that Figure 5 A- Figure 5 D) The PD group showed a significant increase in alpha-synuclein aggregation in the SN and striatum compared with the NC group (P < 0.001), while the SYN group after vitamin intervention showed a significant decrease in alpha-synuclein aggregation compared with the PD group (P < 0.01). Western blotting was used to detect the expression of alpha-synuclein in the SN, and the results showed that Figure 5 E) Consistent with the previous staining results.

[0163] 4. Inflammation and pyroptosis detection

[0164] GFAP and iba-1 are markers of astrocyte and microglial cell activation, representing the inflammatory response in the brain area. Immunohistochemical staining of the SN of mice in each group showed that Figure 6 A- Figure 6 B) The PD group showed an increase in GFAP and iba-1 compared with the NC group (P < 0.001), while the SYN group showed a significant decrease compared with the PD group (P < 0.001). Next, Nissl staining (Nissl staining kit, Beijing Solabio Technology Co., Ltd.) showed that Figure 6 C- Figure 6 F) The PD group showed swollen neuron cell bodies and a decrease in the number of nuclear displacement compared with the NC group (P < 0.001), and the SYN group after vitamin intervention showed a significant improvement compared with the PD group (P < 0.01). Therefore, vitamin intervention significantly improved the main pathological features of PD mice and reduced the inflammatory response.

[0165] Inflammation and pyroptosis play an important role in the development of PD. Western blotting and qPCR (qPCR detection kit, Applied Biological Materials) were used to evaluate the expression of pyroptosis-related proteins and genes in the SN of mice in each group Figure 7 , Figure 8, Figure 9 A- Figure 9 In the PD group, both the protein and mRNA expression levels of pyroptosis pathway-related indicators were significantly increased compared to the NC group (P < 0.05), while the SYN group showed a reversal compared to the PD group (P < 0.05). The mRNA expression levels of inflammatory factors (IL-6, IL-10, TNF-α) were detected by qPCR. Figure 9 C- Figure 9 The results showed that compared with the NC group, the PD group had significantly increased IL-6 and TNF-α (P < 0.05), while the SYN group showed a reversal compared with the PD group (P < 0.05), and the opposite was true for IL-10.

[0166] like Figure 6 Substantia nigra extracted from brain homogenate and analyzed by Western blotting revealed a significant decrease in blood-brain barrier tight junction protein in the PD group (P < 0.05), while the SYN group showed some improvement (P < 0.05). Furthermore, LPS expression in brain homogenate was also examined. Figure 9 The result (F) is consistent with previous results.

[0167] In summary, synbiotic intervention can reduce brain barrier damage in PD mice, decrease LPS in brain tissue, and inhibit the LPS / NLRP3 / pyroptosis pathway and inflammatory response.

[0168] 5. Antioxidant stress detection

[0169] Nrf2 / ARE is a classic antioxidant stress pathway. Western blotting analysis of proteins related to this pathway revealed, for example... Figure 10 , Figure 11 A- Figure 11 In the PD group, the protein level of this pathway was significantly lower than that in the NC group (P < 0.05), while it was significantly higher in the SYN group than in the PD group after synbiotic intervention, indicating that this pathway was significantly activated after intervention (P < 0.05). Detection of key oxidative stress indicators SOD, GSH, and MDA revealed… Figure 11 F- Figure 11 In the PD group, H2O, SOD, and GSH were significantly decreased (P < 0.05), while MDA showed the opposite trend. This phenomenon was reversed after synbiotic intervention. Therefore, synbiotics activate the Nrf2 / ARE antioxidant stress pathway in PD mice.

[0170] 6. Intestinal barrier detection

[0171] Impaired intestinal barrier function may increase the risk of bacterial components entering the bloodstream from the intestinal lumen. LPS can reach the bloodstream via the portal vein, cross the blood-brain barrier, activate inflammatory pathways, and cause brain damage. Figure 12As shown, the colon length in the PD group was significantly shorter, while that in the SYN group was significantly increased after intervention. Quantitative diagrams of colon length for each group are shown below. Figure 12 As shown in E, compared to the PD group, the NC and SYN groups had longer lengths (P < 0.05); Figure 12 As shown in Figure B, HE staining results revealed that the PD group exhibited colonic crypt tortuosity, irregular crypt surfaces, reduced mucus within goblet cells, and basal lymphocyte aggregation. In contrast, after synbiotic intervention, the crypts were neatly arranged, and goblet cell mucus increased. Goblet cells are the main source of intestinal mucus and play a protective role in the intestinal barrier. Goblet cells containing acidic mucin stained blue with albedo blue solution. Figure 12 C- Figure 12 The D-plot showed that, compared with the NC group, the expression of mucin was reduced in the PD group (P < 0.0001), and compared with the PD group, the SYN group showed significant improvement (P < 0.0001).

[0172] Tight junction proteins ZO-1 and Occludin, as essential proteins of the intestinal barrier, accurately reflect the integrity and density of the intestinal barrier. Sections were subjected to antigen retrieval, endogenous peroxidase blockade, and blocking (5% BSA solution, incubation at room temperature for 1 h). Primary antibody was added and the sections were incubated overnight in a chromatography cabinet at 4°C. After washing with PBS, fluorescently labeled secondary antibody was added, and the sections were incubated at 38°C in the dark for 30 min. After washing with PBS, DAPI staining solution was added, and the sections were incubated at room temperature for 10 min. After washing with PBS, the sections were observed and photographed under a fluorescence microscope. Immunofluorescence staining results showed (…). Figure 13 A- Figure 13 In the PD group, compared with the NC group, the levels of ZO-1 and Occludin were significantly decreased (P < 0.001), and the expression of TJ protein was significantly increased after synbiotic intervention (P < 0.01). Similarly, qPCR results showed that synbiotic intervention could increase the mRNA expression of ZO-1 and Occludin (e.g., PD). Figure 13 (E in the middle); then as in Figure 13 The F-value in the figure shows that the increased LPS level in the blood of the PD group was detected by ELISA (P < 0.01), which was reversed after synbiotic intervention (P < 0.0001). Therefore, synbiotic can increase the expression of intestinal tight junction proteins and maintain the integrity of the intestinal barrier.

[0173] 7. Stool testing

[0174] Before the test, mice were fasted for 2 hours and then placed in a clean, transparent, sealed box for another 2 hours. Fecal pellets were collected, and the number of pellets per mouse was counted. Fresh feces were weighed individually, and the wet weight was recorded. The EP tubes containing fresh feces were then placed in an 85°C oven for 24 hours to dry overnight. After drying, the feces were weighed again to obtain the dry weight. The percentage of water content in the feces was calculated based on the difference between the wet and dry weights.

[0175] To evaluate the intestinal motility function of mice, fecal collection was performed for 60 min. Each mouse was assigned to a clean transparent transparent seal box. The number of fecal particles was recorded every 5 min for 60 min.

[0176] The fecal output of the SYN intervention group increased, and the water content of the feces also increased significantly: the PD mouse fecal particles decreased, small and dry, while the SYN intervention group fecal particles increased, large and moist Figure 14 A in Figure 14 C).

[0177] 8. Bacterial flora detection

[0178] One day before the mice were sacrificed, 5-6 fecal particles were collected from each mouse in a sterile 1.5 mL EP tube, the sample was labeled, frozen in liquid nitrogen, and stored with a seal. The whole process was sterile. DNA extraction, library construction, sequencing, and genomic analysis.

[0179] The feces of mice in each group were reserved for metagenomic sequencing (Beijing Yuede Microbial Technology Co., Ltd.), and the results are shown in Figures 15-16 A in Figure 15 A is β diversity, PCA and PcoA dimensionality reduction analysis shows that the overall composition of intestinal microorganisms in each group is significantly different; Figure 15 B is the relative abundance heatmap at the species level, the results show that the PD group of Bacteroides and AKK bacteria shows an increasing trend at the genus and species levels, and the phenomenon is reversed after the intervention of synbiotic, in addition to this, the two species of Deng's bacteria and murine bacillus significantly increase after the intervention of synbiotic.

[0180] Level 2 differential analysis of the KEGG database showed that the PD group significantly increased the species richness of pathways related to environmental information processing, viral diseases, and the body's immune system, while synbiotic intervention increased the species richness of pathways related to cell growth and death. GO database annotation results showed a higher number of genes related to metabolic catalysts and antioxidant activity. pcoa analysis indicated certain differences among groups in the KEGG signaling pathway. Analysis of the eggNOG database to compare the relative gene abundance of the mouse gut microbiota under different functions revealed that the PD group had decreased relative gene abundance in functions such as cell defense mechanisms, signal transduction mechanisms, cell movement, and transcription, while its gene abundance was relatively decreased in functions such as extracellular structure, lipid transport and metabolism, and coenzyme transport and metabolism. Synbiotic intervention showed the opposite trend. pcoa analysis showed significant differences in gene functional diversity among groups. CAZy database analysis revealed that the PD group had increased relative gene abundance in glycoside hydrolases, polysaccharide lysozymes, and carbohydrate esterases, while its relative gene abundance in helper oxidoreductases was decreased, which increased after synbiotic intervention. The above results show that the gut microbiota of PD model mice has undergone significant changes, and synbiotic intervention can reverse this phenomenon.

[0181] Example 4: Synbiotics for the treatment of PD

[0182] 1. Male C57BL / 6J mice (6-8 weeks old) were acclimatized for one week and then randomly divided into three groups of 8 mice each: control group (NC), rotenone model group (PD), and synbiotic intervention group (SYN). At the beginning of the experiment, the PD and SYN groups were administered 30 mg / kg / day of rotenone by gavage in the afternoon, while the NC group was administered an equal volume of solvent (4% chloroform and 0.5% carboxymethyl cellulose CMC) by gavage for 4 weeks. Starting from the fifth week, the SYN group was administered 0.2 mL of synbiotic (containing 9 × 10^9 CFU / day of live bacteria) by gavage in the morning, while the PD and NC groups were administered an equal volume of sterile PBS (0.2 mL). The treatment lasted for two weeks. After the treatment, the mice underwent behavioral tests and were sacrificed. Serum, substantia nigra and striatum brain regions, colon, and fecal particles were collected.

[0183] 2. Behavioral testing

[0184] In a PD model treatment trial, it was found that synbiotics could alleviate behavioral disorders in PD mice. Figure 17 A- Figure 17 The results of the B-rotator experiment showed that, compared with the NC group, the PD group had a significantly increased number of drops (P<0.05) and a significantly reduced total dwell time (P<0.05), while the SYN group after synbiotic intervention showed a significant improvement compared with the PD group (P<0.05). Figure 17 The results of the open field experiment, where C represents the open field, show the same trend as the rotating bar experiment.

[0185] 3. PD brain tissue pathological detection

[0186] As in the preventive intervention experiment, the mouse substantia nigra tissue was immunohistochemically stained, and the tyrosine hydroxylase expression of the PD group mice was significantly reduced compared with the NC group mice (P<0.01), and the SYN group improved this phenomenon (P<0.01) and restored to a similar state to the NC group; Figure 18 ), the PD group of the substantia nigra region of the neuron was shrunk and broken and reduced, and the symptoms of the vitamin intervention group were alleviated (P<0.05), and the vitamin treatment could alleviate the pathological changes of PD.

[0187] 4. Inflammation and cell pyroptosis detection

[0188] The qPCR detection showed that (Fig. 6) Figure 19 Vitamin intervention can alleviate the inflammatory response caused by PD and inhibit the mRNA level expression of pyroptosis-related factors.

[0189] 5. Intestinal barrier detection

[0190] The occurrence of PD will cause a certain degree of damage to the intestinal barrier and increase the intestinal permeability. Through the measurement of the colon length (such as Fig. 7) Figure 20 , it was found that compared with the NC group, the colon length of the PD group mice was shortened to a certain extent (P<0.05), and after the vitamin intervention, it was also increased to a certain extent (P=0.05); through HE staining and Alcian staining (Alcian staining kit, Beijing Solaybao Technology Co., Ltd.) of the colon tissue, it was found that compared with the NC group, the colon basement layer of the PD group was thin, the goblet cells and mucin were reduced, and the crypt arrangement was disordered, and after the vitamin intervention, the SYN group had an improvement effect compared with the PD group.

[0191] Next, through immunofluorescence staining, the tight junction protein of the colon tissue of each group was detected (such as Fig. 8) Figure 21 , the expression of ZO-1 and Occludin of the PD group colon was reduced compared with the NC group (P<0.01), and after the vitamin intervention, the SYN group was increased compared with the PD group (P<0.01), so through the therapeutic intervention using vitamin, the increase of intestinal permeability caused by PD and the increase of intestinal barrier integrity can be alleviated.

[0192] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A composition characterized in that, The composition comprises a complex bacteria powder, the complex bacteria powder comprises Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15, the preservation number of Lactobacillus helveticus LV-20 is CGMCC No: 34563, the preservation number of Lactobacillus plantarum LPP8 is CGMCC No: 34565, and the preservation number of Lactobacillus fermentum PL-15 is CGMCC No: 34564. The live bacteria number ratio of Lactobacillus helveticus LV-20, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 is 0.5-10:0.5-10:

1.

2. The composition of claim 1, wherein, The viable cell count of Lactobacillus helveticus LV-20 strain, Lactobacillus plantarum LPP8 and Lactobacillus fermentum PL-15 in the composite bacterial powder is 1 x 10 5 -10 12 CFU / g.

3. Use of the composition of any one of claims 1-2 in the preparation of a medicament for treating or preventing Parkinson's disease.

4. Use of the composition of any one of claims 1-2 in the preparation of a medicament for treating motor dysfunction caused by Parkinson's disease.

5. Use of the composition of any one of claims 1-2 in the preparation of a medicament for improving brain tissue pathological abnormalities caused by Parkinson's disease.

6. Use of the composition of any one of claims 1-2 in the preparation of a medicament for inhibiting neuroinflammation caused by Parkinson's disease.

7. Use of the composition of any one of claims 1-2 in the preparation of a medicament for treating oxidative stress caused by Parkinson's disease.

8. Use of the composition of any one of claims 1-2 in the preparation of a medicament for repairing intestinal barrier damage caused by Parkinson's disease.

9. Use of the composition of any one of claims 1-2 in the preparation of a medicament for improving constipation caused by Parkinson's disease.

10. Use of the composition of any one of claims 1-2 in the preparation of a medicament for regulating intestinal flora composition changes caused by Parkinson's disease.

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