Preparation method and application of anti-Parkinson hydrogel preparation

Abstract

The invention relates to a preparation method and application of an anti-Parkinson hydrogel preparation, solves the problems of oxidative stress of the brain of Parkinson and dysbacteriosis of intestinal flora, and adopts the technical scheme that the anti-Parkinson hydrogel preparation is prepared by the following steps: extracting bacterial outer membrane vesicles of alcaligenes faecalis through an ultracentrifugation method; the preparation method comprises the following steps: complexing cerium ions and polyphenol substances into a metal polyphenol network, loading the cerium metal polyphenol network into bacterial outer membrane vesicles by a mechanical extrusion method, and finally mixing the substances with inulin to obtain the anti-Parkinson hydrogel preparation. The preparation conditions are mild, the process is simple, the method is stable and reliable, the production cost is low, a new thought for treating the Parkinson's disease is developed, and the traditional Chinese medicine composition is an innovation for treating the Parkinson's disease and has huge economic and social benefits.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a preparation method and application of an anti-Parkinson's hydrogel preparation. Background Art

[0002] Studies have found that the cause of Parkinson's disease may originate in the intestines, and the disease manifests itself in the early stages of intestinal microbial disturbances and gastrointestinal-related symptoms. Oral prebiotics can regulate intestinal flora and mediate the gut-brain axis to play a positive role in Parkinson's treatment. At the same time, the brains of Parkinson's patients are in a state of high oxidative stress, which promotes the occurrence and development of Parkinson's by mediating mitochondrial dysfunction, endoplasmic reticulum dysfunction, etc. At the same time, severe brain inflammation leads to damage to dopaminergic neurons. Therefore, inhibiting brain inflammation and oxidative stress can effectively alleviate Parkinson's disease. Studies have found that polyphenols and cerium ions both have antioxidant / anti-inflammatory effects, and the two can be complexed to form a metal polyphenol network, making them potential nanomaterials for the treatment of Parkinson's.

[0003] Based on the above analysis, if we can address the issue of simultaneously treating the gut and brain, loading the cerium metal polyphenol network into bacterial outer membrane vesicles and then loading the aforementioned substances into a prebiotic hydrogel, they could act on both the gut and brain, achieving simultaneous gut-brain treatment, which would have positive implications for the treatment of Parkinson's disease. However, no relevant reports have been published to date. Summary of the Invention

[0004] In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide a preparation method and application of an anti-Parkinson's hydrogel preparation, which can effectively solve the problems of Parkinson's brain oxidative stress and intestinal flora imbalance.

[0005] To achieve the above objectives, the present invention provides a technical solution, which is a method for preparing an anti-Parkinson's hydrogel preparation. The method comprises extracting bacterial outer membrane vesicles of Alcaligenes faecalis by ultracentrifugation, complexing cerium ions with polyphenols to form a metal polyphenol network, loading the cerium metal polyphenol network into the bacterial outer membrane vesicles by mechanical extrusion, and finally mixing the above substances with inulin to obtain the anti-Parkinson's hydrogel preparation. The method specifically comprises the following steps: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into a tryptone soy broth medium and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 5000-7000 r / min for 10-30 min at 4°C, and the supernatant was removed. The process was repeated twice, and the supernatant was filtered through a 0.45 μm filter membrane to remove residual bacteria. The supernatant was centrifuged at 130,000-150,000 r / min for 3-4 h using an ultracentrifuge. The precipitate was obtained to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis, which were resuspended in sterile PBS at pH 7.4 for later use. The tryptone soy broth medium was composed of: 17.0 g / L tryptone, 3.0 g / L soy peptone, 5.0 g / L sodium chloride, 2.5 g / L potassium dihydrogen phosphate, and 2.5 g / L glucose, with a pH of 7.3±0.2. 2) Preparation of cerium metal polyphenol network: 60-180 mg of cerium ammonium nitrate is dissolved in 10-30 mL of water, and 66-198 mg of polyvinyl pyrrolidone is dissolved in 1-3 mL of water. The two are mixed and stirred in the dark for 0.5-1.5 hours to obtain a reaction solution. 10-30 mg of a polyphenolic substance is weighed and dissolved in 1-3 mL of anhydrous ethanol, and the solution is added to the reaction solution, and the reaction is continued for 8-10 hours. The reaction solution is centrifuged at 8000-10000 r / min for 10-30 minutes, and the precipitate is collected and washed three times with water to obtain a cerium metal polyphenol network. The polyphenolic substance is luteolin, green tea polyphenols, or tannic acid.

[0006] 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) are mixed with the cerium polyphenol nanoparticles prepared in step 2) in a weight-to-volume ratio of 1:1. The suspension is extruded back and forth 10-20 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network; the weight-to-volume ratio is expressed in grams for solids and milliliters for liquids; 4) Preparation of hydrogel formulation Weigh 0.2-0.4 g of inulin and dissolve it in 800-2400 μL of deionized water. Heat the mixture in an 80°C water bath for 5-10 min, then stir it on a thermostatic magnetic stirrer for 5-10 min. After the temperature drops to room temperature, add 1-3 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). Let the mixture stand at room temperature for 18-30 h to obtain an anti-Parkinson's hydrogel preparation.

[0007] Application of the hydrogel preparation prepared by the method in the preparation of an oral anti-Parkinson's agent.

[0008] The hydrogel preparation prepared by the method is used in the preparation of drugs for treating Parkinson's disease.

[0009] The invention has mild preparation conditions, simple process, stable and reliable method, low production cost, opens up new ideas for treating Parkinson's disease, is a major innovation in treating Parkinson's disease, and has huge economic and social benefits. DETAILED DESCRIPTION

[0010] The specific implementation methods of the present invention are described in detail below with reference to the examples.

[0011] The present invention is specifically implemented by the following examples. Example 1

[0012] A method for preparing an anti-Parkinson's hydrogel preparation comprises the following steps: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into tryptone soy broth and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 5000 rpm for 10 min at 4°C, and the supernatant was removed. This was repeated twice and filtered through a 0.45 μm filter to remove residual bacteria. The supernatant was then centrifuged at 130,000 rpm for 3 h in an ultracentrifuge. The precipitate was collected to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis and resuspended in sterile PBS at pH 7.4 for later use. 2) Preparation of cerium metal polyphenol network: Weigh 60 mg of cerium ammonium nitrate and dissolve it in 10 mL of water. Dissolve 66 mg of polyvinyl pyrrolidone in 1 mL of water. Mix the two and react with stirring in the dark for 0.5 h to obtain a reaction solution. Weigh 10 mg of luteolin and dissolve it in 1 mL of anhydrous ethanol. Add this to the reaction solution and continue the reaction for 8 h. Centrifuge the reaction solution at 8000 rpm for 10 min, collect the precipitate, and wash it three times with water to obtain the cerium metal polyphenol network. 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) were mixed with the cerium polyphenol nanoparticles prepared in step 2) at a weight-to-volume ratio of 1:1. The suspension was extruded back and forth 10 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network. 4) Preparation of hydrogel formulation Weigh 0.2 g of inulin and dissolve it in 800 μL of deionized water. Heat the mixture in an 80°C water bath for 5 min, then stir it on a thermostatic magnetic stirrer for 5 min. After the temperature drops to room temperature, add 1 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). Let the mixture stand at room temperature for 18 h to obtain an anti-Parkinson's hydrogel preparation. Example 2

[0013] A method for preparing an anti-Parkinson's hydrogel preparation comprises the following steps: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into tryptone soy broth and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 6000 rpm for 20 min at 4°C, and the supernatant was removed. This was repeated twice and filtered through a 0.45 μm filter to remove residual bacteria. The supernatant was then centrifuged at 140,000 rpm for 3.5 h in an ultracentrifuge. The precipitate was collected to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis and resuspended in sterile PBS at pH 7.4 for later use. 2) Preparation of cerium metal polyphenol network: Weigh 120 mg of cerium ammonium nitrate and dissolve it in 20 mL of water. Dissolve 132 mg of polyvinyl pyrrolidone in 2 mL of water. Mix the two and stir in the dark for 1 hour to obtain a reaction solution. Weigh 20 mg of green tea polyphenols and dissolve them in 2 mL of anhydrous ethanol. Add this to the reaction solution and continue the reaction for 9 hours. Centrifuge the reaction solution at 9000 rpm for 20 minutes, collect the precipitate, and wash it three times with water to obtain a cerium metal polyphenol network. 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) were mixed with the cerium polyphenol nanoparticles prepared in step 2) at a weight-to-volume ratio of 1:1. The suspension was extruded back and forth 15 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network. 4) Preparation of hydrogel formulation Weigh 0.3 g of inulin and dissolve it in 1600 μL of deionized water. Heat the mixture in an 80°C water bath for 7.5 min, then stir it on a thermostatic magnetic stirrer for 7.5 min. After the temperature drops to room temperature, add 2 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). The mixture is allowed to stand at room temperature for 24 h to obtain an anti-Parkinson's hydrogel preparation. Example 3

[0014] A method for preparing an anti-Parkinson's hydrogel preparation comprises the following steps: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into tryptone soy broth and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 7,000 rpm for 30 min at 4°C, and the supernatant was removed. This was repeated twice and filtered through a 0.45 μm filter to remove residual bacteria. The supernatant was then centrifuged at 150,000 rpm for 4 h in an ultracentrifuge. The precipitate was collected to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis and resuspended in sterile PBS at pH 7.4 for later use. 2) Preparation of cerium metal polyphenol network: Weigh 180 mg of cerium ammonium nitrate and dissolve it in 30 mL of water. Dissolve 198 mg of polyvinyl pyrrolidone in 3 mL of water. Mix the two and stir in the dark for 1.5 h to obtain a reaction solution. Weigh 30 mg of tannic acid and dissolve it in 3 mL of anhydrous ethanol. Add this to the reaction solution and continue the reaction for 10 h. Centrifuge the reaction solution at 10,000 rpm for 30 min, collect the precipitate, and wash it three times with water to obtain a cerium metal polyphenol network. 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) were mixed with the cerium polyphenol nanoparticles prepared in step 2) at a weight-to-volume ratio of 1:1. The suspension was extruded back and forth 20 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network. 4) Preparation of hydrogel formulation Weigh 0.4 g of inulin and dissolve it in 2400 μL of deionized water. Heat the mixture in an 80°C water bath for 10 min, then stir it on a thermostatic magnetic stirrer for 10 min. After the temperature drops to room temperature, add 3 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). Let the mixture stand at room temperature for 30 h to obtain an anti-Parkinson's hydrogel preparation.

[0015] The hydrogel preparation prepared by the present invention is stable and reliable, has the effect of regulating the intestines and controlling the brain, and thus treats Parkinson's disease. Scientific experiments have verified that the effect is very good. Taking Example 1 as an example, the relevant experimental data are as follows: 1. 16S rDNA Observation of Changes in Mouse Intestinal Microbiota First, a Parkinson's mouse model was established by intraperitoneal injection of MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine). After the Parkinson's mouse model was successfully established, the mice were given the drug. The animal pharmacodynamics experiment was divided into three groups: (1) normal mice given saline; (2) MPTP-induced Parkinson's mice given saline; and (3) MPTP-induced Parkinson's mice given the hydrogel formulation. The experimental mice were given the drug by gavage once a day for two consecutive weeks.

[0016] After treatment, feces were collected from each group of mice and sent to Shanghai Meiji Biotechnology Co., Ltd. on dry ice for 16S rDNA sequencing. The experimental data were analyzed for community composition at the phylum and genus levels to examine changes in the composition of the mouse intestinal microbiota.

[0017] Through phylum-level community analysis, the changes in the composition of the intestinal flora of mice in different groups were examined. The results showed that compared with the control group, the relative abundance of Firmicutes in the intestines of MPTP-induced Parkinson's mice increased, while the relative abundance of Bacteroidetes and Campylobacter decreased, indicating that the intestinal flora of Parkinson's mice had undergone significant changes. After treatment with the hydrogel preparation, the imbalance of the intestinal flora of Parkinson's mice could be reversed to varying degrees, reducing the relative abundance of Firmicutes that increased after MPTP treatment, while increasing the relative abundance of Bacteroidetes. The phylum-level community analysis proved that the final preparation had the effect of regulating the composition of the intestinal flora of Parkinson's mice.

[0018] Genus-level community analysis examined changes in the intestinal microbiome composition of mice in different groups. The results showed that compared to the control group, the abundance of SCFA-producing microorganisms in the intestinal tract of the MPTP group decreased, and this decrease in the abundance of these microorganisms further exacerbated Parkinson's disease pathological symptoms. After the hydrogel preparation was treated in Parkinson's mice, the abundance of harmful bacteria decreased, while the abundance of beneficial bacteria increased, indicating that the hydrogel preparation has a good ability to regulate the intestinal microbiome and can be used to treat Parkinson's disease.

[0019] 2. ELISA experiment to investigate the expression level of inflammatory cytokines in mouse brain First, a Parkinson's mouse model was established by intraperitoneal injection of MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine). After the Parkinson's mouse model was successfully established, the mice were given the drug. The animal pharmacodynamics experiments were divided into three groups: (1) normal mice given saline; (2) MPTP-induced Parkinson's mice given saline; and (3) MPTP-induced Parkinson's mice given the hydrogel formulation.

[0020] After the pharmacodynamics experiments, the substantia nigra of the mouse brain was isolated, cleaned, and homogenized in pre-chilled PBS using a cryogenic tissue grinder. The suspension was centrifuged at 3000 rpm for 15 minutes at 4°C, and the supernatant was collected. Tumor necrosis factor (TNF-α) and interleukin (IL-1β) levels in the substantia nigra were quantified using a BCA quantification kit and then measured according to the ELISA kit instructions.

[0021] Results showed that compared with the control group, the MPTP group showed significantly increased levels of pro-inflammatory cytokines TNF-α and IL-1β, indicating neuroinflammation and a dysregulation of the brain's immune microenvironment. This phenomenon was reversed after treatment with the hydrogel formulation, demonstrating its potent anti-inflammatory effect.

[0022] At the same time, while the present invention conducted the above experiment on Example 1, the same experiment was also conducted on Examples 2-3, and the same or similar results as those in Example 1 were obtained. They are not listed here one by one, which shows that the present invention has good effects and stable and reliable therapeutic effects.

[0023] The above experiments show that the method of the present invention is stable and reliable, can solve the problems of brain oxidative stress and intestinal flora imbalance, and has the following outstanding beneficial technical effects compared with the existing technology: 1) The hydrogel preparation of the present invention has synergistic antioxidant / anti-inflammatory effects to alleviate Parkinson's disease; 2) The hydrogel preparation of the present invention exerts therapeutic functions in both the intestine and the brain, achieving gut-brain linkage through the dual effects of regulating the intestine and controlling the brain. This is a major innovation in drugs for the treatment of Parkinson's disease, with huge economic and social benefits.

[0024] It should be pointed out that the above is only a preferred embodiment of the present invention and does not impose any form of limitation on the present invention. Any technician familiar with this profession can make changes or modify the technical content disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention, and all of them fall within the scope of protection of the present invention.

Claims

1. A method for preparing an anti-Parkinson's hydrogel preparation, characterized in that: The following steps are involved: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into a tryptone soy broth medium and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 5000-7000 r / min for 10-30 min at 4°C, and the supernatant was removed. The process was repeated twice, and the supernatant was filtered through a 0.45 μm filter membrane to remove residual bacteria. The supernatant was centrifuged at 130,000-150,000 r / min for 3-4 h using an ultracentrifuge. The precipitate was obtained to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis, which were resuspended in sterile PBS at pH 7.4 for later use. The tryptone soy broth medium was composed of: 17.0 g / L tryptone, 3.0 g / L soy peptone, 5.0 g / L sodium chloride, 2.5 g / L potassium dihydrogen phosphate, and 2.5 g / L glucose, with a pH of 7.3±0.

2. 2) Preparation of cerium metal polyphenol network: 60-180 mg of cerium ammonium nitrate is dissolved in 10-30 mL of water, and 66-198 mg of polyvinyl pyrrolidone is dissolved in 1-3 mL of water. The two are mixed and stirred in the dark for 0.5-1.5 hours to obtain a reaction solution. 10-30 mg of a polyphenolic substance is weighed and dissolved in 1-3 mL of anhydrous ethanol, and the mixture is added to the reaction solution, and the reaction is continued for 8-10 hours. The reaction solution is centrifuged at 8000-10000 r / min for 10-30 minutes, and the precipitate is collected and washed three times with water to obtain a cerium metal polyphenol network. The polyphenolic substance is luteolin, green tea polyphenols, or tannic acid. 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) are mixed with the cerium polyphenol nanoparticles prepared in step 2) in a weight-to-volume ratio of 1:

1. The suspension is extruded back and forth 10-20 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network; the weight-to-volume ratio is expressed in grams for solids and milliliters for liquids; 4) Preparation of hydrogel formulation Weigh 0.2-0.4 g of inulin and dissolve it in 800-2400 μL of deionized water. Heat the mixture in an 80°C water bath for 5-10 min, then stir it on a thermostatic magnetic stirrer for 5-10 min. After the temperature drops to room temperature, add 1-3 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). Let the mixture stand at room temperature for 18-30 h to obtain an anti-Parkinson's hydrogel preparation.

2. The method for preparing the anti-Parkinson's hydrogel preparation according to claim 1, characterized in that: The following steps are involved: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into tryptone soy broth and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 5000 rpm for 10 min at 4°C, and the supernatant was removed. This was repeated twice and filtered through a 0.45 μm filter to remove residual bacteria. The supernatant was then centrifuged at 130,000 rpm for 3 h in an ultracentrifuge. The precipitate was collected to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis and resuspended in sterile PBS at pH 7.4 for later use. 2) Preparation of cerium metal polyphenol network: Weigh 60 mg of cerium ammonium nitrate and dissolve it in 10 mL of water. Dissolve 66 mg of polyvinyl pyrrolidone in 1 mL of water. Mix the two and react with stirring in the dark for 0.5 h to obtain a reaction solution. Weigh 10 mg of luteolin and dissolve it in 1 mL of anhydrous ethanol. Add this to the reaction solution and continue the reaction for 8 h. Centrifuge the reaction solution at 8000 rpm for 10 min, collect the precipitate, and wash it three times with water to obtain the cerium metal polyphenol network. 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) were mixed with the cerium polyphenol nanoparticles prepared in step 2) at a weight-to-volume ratio of 1:

1. The suspension was extruded back and forth 10 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network. 4) Preparation of hydrogel formulation Weigh 0.2 g of inulin and dissolve it in 800 μL of deionized water. Heat the mixture in an 80°C water bath for 5 min, then stir it on a thermostatic magnetic stirrer for 5 min. After the temperature drops to room temperature, add 1 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). Let the mixture stand at room temperature for 18 h to obtain an anti-Parkinson's hydrogel preparation.

3. The method for preparing the anti-Parkinson's hydrogel preparation according to claim 1, characterized in that: The following steps are involved: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into tryptone soy broth and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 6000 rpm for 20 min at 4°C, and the supernatant was removed. This was repeated twice and filtered through a 0.45 μm filter to remove residual bacteria. The supernatant was then centrifuged at 140,000 rpm for 3.5 h in an ultracentrifuge. The precipitate was collected to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis and resuspended in sterile PBS at pH 7.4 for later use. 2) Preparation of cerium metal polyphenol network: Weigh 120 mg of cerium ammonium nitrate and dissolve it in 20 mL of water. Dissolve 132 mg of polyvinyl pyrrolidone in 2 mL of water. Mix the two and stir in the dark for 1 hour to obtain a reaction solution. Weigh 20 mg of green tea polyphenols and dissolve them in 2 mL of anhydrous ethanol. Add this to the reaction solution and continue the reaction for 9 hours. Centrifuge the reaction solution at 9000 rpm for 20 minutes, collect the precipitate, and wash it three times with water to obtain a cerium metal polyphenol network. 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) were mixed with the cerium polyphenol nanoparticles prepared in step 2) at a weight-to-volume ratio of 1:

1. The suspension was extruded back and forth 15 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network. 4) Preparation of hydrogel formulation Weigh 0.3 g of inulin and dissolve it in 1600 μL of deionized water. Heat the mixture in an 80°C water bath for 7.5 min, then stir it on a thermostatic magnetic stirrer for 7.5 min. After the temperature drops to room temperature, add 2 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). The mixture is allowed to stand at room temperature for 24 h to obtain an anti-Parkinson's hydrogel preparation.

4. The method for preparing the anti-Parkinson's hydrogel preparation according to claim 1, characterized in that: The following steps are involved: 1) Extraction of bacterial outer membrane vesicles from Alcaligenes faecalis: Alcaligenes faecalis was inoculated into tryptone soy broth and cultured aerobically in a 37°C incubator. The bacterial solution was centrifuged at 7,000 rpm for 30 min at 4°C, and the supernatant was removed. This was repeated twice and filtered through a 0.45 μm filter to remove residual bacteria. The supernatant was then centrifuged at 150,000 rpm for 4 h in an ultracentrifuge. The precipitate was collected to obtain bacterial outer membrane vesicles derived from Alcaligenes faecalis and resuspended in sterile PBS at pH 7.4 for later use. 2) Preparation of cerium metal polyphenol network: Weigh 180 mg of cerium ammonium nitrate and dissolve it in 30 mL of water. Dissolve 198 mg of polyvinyl pyrrolidone in 3 mL of water. Mix the two and stir in the dark for 1.5 h to obtain a reaction solution. Weigh 30 mg of tannic acid and dissolve it in 3 mL of anhydrous ethanol. Add this to the reaction solution and continue the reaction for 10 h. Centrifuge the reaction solution at 10,000 rpm for 30 min, collect the precipitate, and wash it three times with water to obtain a cerium metal polyphenol network. 3) Preparation of bacterial outer membrane vesicles loaded with cerium metal polyphenol network: The outer membrane vesicles derived from Alcaligenes faecalis prepared in step 1) were mixed with the cerium polyphenol nanoparticles prepared in step 2) at a weight-to-volume ratio of 1:

1. The suspension was extruded back and forth 20 times using a liposome extruder containing polycarbonate membranes with pore sizes of 0.8 μm and 0.4 μm, respectively, to obtain bacterial outer membrane vesicles loaded with a cerium polyphenol network. 4) Preparation of hydrogel formulation Weigh 0.4 g of inulin and dissolve it in 2400 μL of deionized water. Heat the mixture in an 80°C water bath for 10 min, then stir it on a thermostatic magnetic stirrer for 10 min. After the temperature drops to room temperature, add 3 ml of the bacterial outer membrane vesicles loaded with cerium metal polyphenol network prepared in step 3). Let the mixture stand at room temperature for 30 h to obtain an anti-Parkinson's hydrogel preparation.

5. Use of the hydrogel preparation prepared by the method according to any one of claims 1 to 4 in the preparation of an oral anti-Parkinson's agent.

6. Use of the hydrogel preparation prepared by the method according to any one of claims 1 to 4 in preparing drugs for treating Parkinson's disease.