Application of combination of achyranthes bidentata polypeptide and human umbilical cord mesenchymal stem cell exosome in preparation of medicine for preventing and / or treating Parkinson's disease

Abstract

The invention belongs to the technical field of biological medicines, and relates to application of achyranthes bidentata polypeptide combined with human umbilical cord mesenchymal stem cell exosome in preparation of a medicine for preventing and / or treating Parkinson's disease, the achyranthes bidentata polypeptide combined with the human umbilical cord mesenchymal stem cell exosome is used for treating mice, and 1-methyl-4-phenyl-1, 2, 3, 4-tetramethyl-1, 3, 4-tetramethyl-1, 3, 4-tetramethyl-1, 3, 4-tetramethyl-1, 3, 4-tetramethyl-1, 3, 4-tetramethyl-1, 3, 4 The method comprises the following steps: carrying out Parkinson model (PD) modeling on a mouse by using 2, 3, 4, 6-tetrahydropyridine (MPTP); it is found that the treatment can significantly relieve PD related symptoms, for example, the number of dopaminergic neurons in a nigra part is recovered, expression of tyrosine hydroxylase in a middle brain part is improved, movement and olfactory dysfunction of a PD model mouse is relieved, activation of olfactory bulbs and microglia and astrocytes in the middle brain part is reduced, and activity of neurons in the olfactory bulbs is improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and discloses an application of Achyranthes bidentata polypeptide combined with human umbilical cord mesenchymal stem cell exosomes in the preparation of a drug for preventing and / or treating Parkinson's disease. Background Art

[0002] Parkinson's disease (PD), also known as paralysis agitans, is the second largest neurodegenerative disease after Alzheimer's disease that seriously affects the quality of human life. According to statistics, one in every 800 people in the world suffers from Parkinson's disease. By 2030, due to the acceleration of the aging process, the prevalence of Parkinson's disease will double, and it is expected to exceed 9 million patients. Parkinson's disease is characterized by progressiveness, multiple occurrences, and insidious onset, and is mainly manifested by slow movement, muscle rigidity, resting tremor, and postural instability. The main pathological characteristics of Parkinson's disease are mainly manifested in the reduction of dopaminergic neurons in the substantia nigra. The cells that produce dopamine in the brain gradually lose the function of affecting the nervous system, which limits the patient's ability to control muscles. The clinical treatment of Parkinson's disease is mainly based on dopamine replacement therapy. Long-term use of this therapy can cause a variety of adverse reactions, such as mental symptoms such as restlessness, insomnia, and hallucinations. Therefore, finding new PD treatment drugs has huge economic benefits and social value.

[0003] Achyranthes bidentata is the dried root of Achyranthes bidentata BI., a plant of the Amaranthaceae family. It is a commonly used Chinese medicine with physiological functions such as removing blood stasis and promoting menstruation, nourishing the liver and kidneys, strengthening tendons and bones, promoting diuresis and relieving stranguria, and guiding blood downward. Achyranthes bidentata polypeptide is a biologically active substance isolated from Achyranthes bidentata. A purified Achyranthes bidentata polypeptide component was obtained by further purification through high performance liquid chromatography (HPLC).

[0004] Human umbilical cord mesenchymal stem cells (hUCMCSs) are derived from human umbilical cord tissue and have the characteristics of easy extraction, strong self-renewal ability, low immunogenicity and the ability to secrete a variety of functional factors. Therefore, human umbilical cord mesenchymal stem cells are considered to be the first choice for clinical stem cell therapy and are widely used in a variety of tissue repair. Exosomes are a lipid bilayer vesicle with a diameter of 50-150 nm. They are produced and excreted from cells through active secretion. Exosomes contain a variety of substances, including lipid molecules, proteins and nucleic acids (including DNA, RNA and various non-coding RNAs). Exosomes play an extremely important role in information exchange and material transfer between cells. At present, more and more studies have shown that stem cell exosomes have physiological activities similar to those of stem cells.

[0005] The nasal cavity is rich in capillary lymphatic vessels, and the surface of cells in the respiratory zone has a large number of tiny villi, which is an ideal mucosal drug administration route. The nasal mucosa has high permeability and relatively few enzymes, and its decomposition effect on protein drugs is lower than that of the gastrointestinal mucosa. In addition, nasal administration can enter the brain along the olfactory nerve through the bipolar olfactory cells of the nasal olfactory epithelium, avoiding the barrier effect of the blood-brain barrier on protein drugs. Therefore, nasal administration is a very effective drug administration method that acts on the central nervous system. Summary of the invention

[0006] In view of the shortcomings of the prior art, the present invention provides the use of Achyranthes bidentata polypeptide combined with human umbilical cord mesenchymal stem cell exosomes in the preparation of a drug for preventing and / or treating Parkinson's disease. It was found that Achyranthes bidentata polypeptide combined with human umbilical cord mesenchymal stem cell exosomes can restore the number of dopaminergic neurons in the substantia nigra, increase the expression of tyrosine hydroxylase in the midbrain, alleviate the motor and olfactory dysfunction of PD model mice, reduce the activation of microglia and astrocytes in the olfactory bulb and midbrain, and increase the activity of neurons in the olfactory bulb.

[0007] The technical solution provided by the present invention is as follows:

[0008] The present invention provides use of Achyranthes bidentata polypeptide combined with human umbilical cord mesenchymal stem cell exosomes in the preparation of a medicament for preventing and / or treating Parkinson's disease.

[0009] Furthermore, the drug includes 1~2×10 8 / kg human umbilical cord mesenchymal stem cell exosomes and 50~100 μg / kg Achyranthes bidentata peptides.

[0010] Furthermore, the Achyranthes bidentata polypeptide is prepared by the following method: Cutting the dried root of Achyranthes bidentata into pieces, mixing the cut dried root of Achyranthes bidentata with water and then steaming to obtain a mixed liquid; Adding ammonium sulfate to the mixed solution for precipitation, centrifuging, and freeze-drying to obtain a crude extract of Achyranthes bidentata polypeptide; The crude extract of Achyranthes bidentata polypeptide is dissolved in water to prepare a solution, and then the solution is purified by high performance liquid chromatography to obtain the Achyranthes bidentata polypeptide.

[0011] Furthermore, the cooking temperature is 80-85°C and the cooking time is 40-60 minutes.

[0012] Furthermore, the conditions of the HPLC are as follows: C18 reverse phase column, mobile phase is water / acetonitrile containing 0.1% trifluoroacetic acid, the volume ratio of water to acetonitrile is 4:1, flow rate is 1.0 mL / min, and the wavelength is 220 nm UV detector.

[0013] Furthermore, the human umbilical cord mesenchymal stem cell exosomes are prepared by the following method: The primary human umbilical cord mesenchymal stem cells were subcultured in a serum-containing medium. When the density of the third generation human umbilical cord mesenchymal stem cells reached 80%, the medium was changed to a serum-free medium and cultured for 48 hours, and the supernatant was collected. The supernatant was then centrifuged at 400-500 g to remove cells and cell debris, and then filtered through a 0.22 μm filter; The filtered supernatant is treated with an exosome separation kit, and the umbilical cord mesenchymal stem cell exosomes are obtained by separation and purification through a specific buffer and centrifugation step.

[0014] Furthermore, the application includes any of the following: Improve motor dysfunction and olfactory perception; Increase the number of dopaminergic neurons in the brain; Inhibit the abnormal activation of microglia and astrocytes in the brain; Enhances the activity of olfactory bulb neurons.

[0015] Furthermore, the drug is administered via nasal cavity administration.

[0016] Furthermore, the medicine is a solution.

[0017] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0018] Beneficial Effects

[0019] The present invention uses Achyranthes bidentata polypeptide combined with human umbilical cord mesenchymal stem cell exosomes to treat mice, and uses 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to model Parkinson's disease (PD) in mice. It is found that the treatment can significantly alleviate PD-related symptoms, such as restoring the number of dopaminergic neurons in the substantia nigra, increasing the expression of tyrosine hydroxylase in the midbrain, alleviating motor and olfactory dysfunction in PD model mice, reducing the activation of microglia and astrocytes in the olfactory bulb and midbrain, and increasing the activity of neurons in the olfactory bulb.

[0020] The drug can be administered through the nasal cavity and enter the brain along the olfactory nerve through the bipolar olfactory cells of the nasal olfactory epithelium, thereby avoiding the barrier effect of the blood-brain barrier on protein drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Preparation of Achyranthes bidentata polypeptide. A: Preparation process; B: HPLC purification; C: Achyranthes bidentata polypeptide after freeze-drying.

[0022] Figure 2 Electron microscopic image of exosomes from human umbilical cord mesenchymal stem cells. Arrows indicate exosomes. Scale bar: 200 nm.

[0023] Figure 3 (A) PD model mouse modeling and drug intervention process. ip: intraperitoneal injection; in: nasal administration; (B) pole climbing test; (C) olfactory test. *** p<0.001, compared with the control group; # p<0.05, ## p<0.01, ### p<0.001, compared with the MPTP group; &p<0.05, compared with the MPTP+Achyranthes bidentata polypeptide group; $ p<0.05, compared with the MPTP+exosome group.

[0024] Figure 4 (A) Immunofluorescence detection showed that Achyranthes bidentata polypeptide combined with human umbilical cord stem cell mesenchymal exosomes increased the expression of tyrosine hydroxylase (TH) in the substantia nigra of PD model mice; scale bar: 50 mm. (B) Quantitative analysis of the number of TH-positive cells in the substantia nigra, *** p<0.001, compared with the control group; ## p<0.01, ### p<0.001, compared with the MPTP group;&&p<0.01, compared with the MPTP+Achyranthes bidentata polypeptide group;$ p<0.05, compared with the MPTP+exosome group.

[0025] Figure 5 (A) Immunofluorescence detection showed that Achyranthes bidentata polypeptide combined with human umbilical cord stem cell mesenchymal exosomes reduced the expression of Iba-1 in the substantia nigra of PD model mice; scale bar: 50 mm. (B) Quantitative analysis of Iba-1 fluorescence intensity in the substantia nigra, ** p<0.01, compared with the control group; # p<0.05, compared with the MPTP group; & p<0.05, compared with the MPTP+Achyranthes bidentata polypeptide group; $ p<0.05, compared with the MPTP+exosome group.

[0026] Figure 6 (A) Immunofluorescence detection showed that Achyranthes bidentata polypeptide combined with human umbilical cord stem cell mesenchymal exosomes reduced the expression of GFAP in the substantia nigra of PD model mice; scale bar: 50 mm. (B) Quantitative analysis of GFAP fluorescence intensity in the substantia nigra, *** p<0.001, compared with the control group; # p<0.05, ## p <0.01, compared with the MPTP group; &p<0.05, compared with the MPTP+Achyranthes bidentata polypeptide group; $ p<0.05, compared with the MPTP+exosome group.

[0027] Figure 7Immunofluorescence detection showed that Achyranthes bidentata polypeptide combined with human umbilical cord stem cell mesenchymal exosomes increased the expression of Phospho-S6 Ribosomal protein in the lateral and central regions of the olfactory bulb of PD model mice; red is Phospho-S6 Ribosomal protein; DAPI (blue) shows the cell nucleus. Scale bar: 25 mm. *** p<0.001, compared with the control group; #p<0.05, # p<0.01, ### p<0.001, compared with the MPTP group; &p<0.05, compared with the MPTP+Achyranthes bidentata polypeptide group; $ p<0.05, compared with the MPTP+exosome group.

[0028] Figure 8 Immunofluorescence detection showed that Achyranthes bidentata polypeptide combined with human umbilical cord stem cell mesenchymal exosomes reduced the expression of Iba-1 (red) in the lateral and central regions of the olfactory bulb of PD model mice; DAPI (blue) indicated the cell nucleus. Scale bar: 25 mm. ***p<0.001, compared with the control group; # p<0.05, # p<0.01, ### p<0.001, compared with the MPTP group; &p<0.05, compared with the MPTP+Achyranthes bidentata polypeptide group; $ p<0.05, compared with the MPTP+exosome group.

[0029] Fig. 9 Immunofluorescence detection showed that Achyranthes bidentata polypeptide combined with human umbilical cord stem cell mesenchymal exosomes reduced the expression of GFAP (red) in the lateral region of the olfactory bulb of PD model mice; DAPI (blue) indicated the cell nucleus. Scale bar: 25 mm. ** p<0.01, compared with the control group; # p<0.05, ## p<0.01, compared with the MPTP group; &p<0.05, compared with the MPTP+Achyranthes bidentata polypeptide group; $ p<0.05, compared with the MPTP+exosome group. DETAILED DESCRIPTION

[0030] The present invention is further described in detail below in conjunction with the accompanying drawings:

[0031] The experimental methods used in the embodiments of the present invention are conventional methods unless otherwise specified.

[0032] The reagents and materials used in this example can all be purchased conventionally. The quantitative experiments involved in the examples were repeated at least three times, and the results were averaged.

[0033] The dried roots of Achyranthes bidentata were purchased from Yongjitang Pharmacy.

[0034] The primary human umbilical cord mesenchymal stem cells used were purchased from Nanjing Taisheng Biotechnology Co., Ltd.

[0035] The C57BL / 6J mice used were purchased from the Experimental Animal Center of Nantong University.

[0036] The 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) used was purchased from Sigma-Aldrich Company with the product number M0896.

[0037] Example 1

[0038] 1. Preparation of Achyranthes bidentata polypeptide ( Figure 1 )

[0039] The dried roots of Achyranthes bidentata were cut into pieces and decocted in a water bath (temperature 80-85°C, time 40-60 min). The aqueous extract was precipitated with ammonium sulfate twice and centrifuged at high speed. The precipitate was freeze-dried to obtain the crude extract of Achyranthes bidentata polypeptide. The obtained crude extract of Achyranthes bidentata polypeptide was dissolved in water and further purified by high performance liquid chromatography (HPLC). The chromatographic conditions were as follows: C18 reverse phase column, mobile phase was water / acetonitrile (4:1, volume ratio) containing 0.1% trifluoroacetic acid, flow rate was 1.0 mL / min, and UV detector (220 nm).

[0040] 2. Preparation of exosomes from human umbilical cord mesenchymal stem cells ( Figure 2 )

[0041] The cells were cultured in DMEM / F12 (Corning) containing 10% fetal bovine serum and placed in a cell culture incubator at 37°C and 5% carbon dioxide saturated humidity. When the cell density reached 80%, they were digested with 0.25% trypsin (Gibco) containing EDTA and subcultured. When the density of human umbilical cord mesenchymal stem cells at passage 3 reached 80%, the culture medium was changed to serum-free medium and cultured for 48 h. The supernatant was collected and continuously ultracentrifuged at 400 g for 10 min to remove dead cells and cell debris, and then filtered through a 0.22 μm filter (Millipore). Exosomes were isolated from 15 ml of cell culture supernatant using the exoEasy Kit (QIAGEN). The filtered supernatant was transferred to a new tube, and an equal volume of buffer XBP was added to the supernatant. Then, a total of 30 ml of the mixture was added to the exoEasy spin column and centrifuged at 500 g for 1 min. After discarding the liquid, the spin column was returned to the same collection tube. Repeat the above steps until no supernatant remains. Then, add 10 ml of Buffer XWP to the spin column and centrifuge at 5000 g for 5 min to remove the remaining buffer. Transfer the spin column to a new collection tube. Add 400 μl of Buffer XE to the membrane and incubate for 1 min. Collect the eluate by centrifugation at 500 g for 5 min. Add the eluate to the spin column and incubate again for 1 min. Finally, collect the eluate after centrifugation at 5000 g for 5 min and store at -80°C until use.

[0042] Example 2

[0043] 1. Preparation of Parkinson's disease animal model and drug administration

[0044] 8-week-old C57BL / 6J male mice were injected intraperitoneally with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (N-methyl-4-phenyl-l,2,3,6-tetrahydropyridine, MPTP) to induce PD model mice. Before treatment, the mice were trained in related behavioral training such as climbing poles for 3 days. To avoid the influence of individual differences, mice with approximately the same level of motor ability were selected and divided into a control group and a model group. Then, the model group mice were injected intraperitoneally with MPTP (25 mg / kg / day) for one week, and then behavioral evaluation was performed ( Figure 3 A). Compared with the control group mice (injected with the same volume of saline), mice with significantly decreased behavior were considered to be successfully induced PD model mice and used for subsequent experiments.

[0045] The successful PD model mice were divided into four groups, namely, MPTP group, MPTP+Achyranthes bidentata polypeptide group, MPTP+exosome group, and MPTP+Achyranthes bidentata polypeptide / exosome group. The mice were treated by nasal administration. Achyranthes bidentata polypeptide and exosomes were dissolved in PBS solution, and the exosome dose was 2×10 8 The dosage of Achyranthes bidentata polypeptide was 100 μg / kg, and the drug was given once every 3 days, for a total of 4 times. The mice in the MPTP group were treated in the same way and given the same volume of PBS solution.

[0046] 2. Pole climbing experiment

[0047] Make a rod with a diameter of 1 cm and a length of 50 cm, with a cork ball with a diameter of 1.5 cm fixed on the top, and wrap it with gauze to increase friction. Place the rod vertically, put the mouse on the ball at the top of the homemade climbing rod, and then record the time from the start of the movement to the complete head down until the animal returns to the bottom of the rod. Each test is 5 minutes apart, and the average value is taken for a total of 3 tests. Three days before the tail vein injection, the mice were trained to climb the rod at the same time every day, and the mice with large differences were eliminated to reduce experimental errors.

[0048] 3. Olfactory experiment

[0049] The mice were fasted for 24 hours in advance. A small piece of cheese was placed during the fasting period for the mice to get familiar with the smell and taste so that they could find it in the experiment. A clean cage (42 cm long, 24 cm wide, 15 cm high) was prepared and cheese was buried in the middle, upper left, lower right, upper right, and lower left of the clean bedding. The cheese should be buried 1 cm below the bedding. The animal was gently placed in the center of the experimental cage, and the time it took for the mouse to find the cheese was recorded. If it was not found within 300 s, it was recorded as 300 s. Statistical analysis was performed after removing the minimum and maximum values.

[0050] 4. Immunofluorescence analysis of substantia nigra and olfactory bulb

[0051] The experimental steps are as follows: (1) The mouse brain fixed with 4% paraformaldehyde overnight was dehydrated with 20% and 30% sucrose at 4°C, and then removed after being completely dehydrated and sinking to the bottom; (2) After the brain block was embedded in OCT, continuous coronal sections of the substantia nigra of the mouse midbrain were prepared on a freezing microtome with a thickness of 12 μm. The sections were dried at 37°C for later use; (3) The slides were placed in 0.01M PBS and washed three times at room temperature for 10 min each time; (4) A blocking solution containing 5% BSA and 0.5% Triton X-100 was added and blocked for 1 h at room temperature; (5) The blocking solution was discarded and the primary antibody diluted in proportion was added dropwise for incubation at 4°C overnight. Commonly used primary antibody ratios: Chicken anti-TH (1:500, ab76442, Abcam), Rabbit anti-Iba-1 (1:500, ab7260, Abcam), Mouse anti-GFAP (1:500, MAB360, Millipore), Rabbit anti-pS6 Ribosomal Protein (1:50, 4858S, Cell Signaling Technology); (6) Wash three times with 0.01M PBS at room temperature, 10 min each time; (7) Add secondary antibodies diluted in 0.01M PBS according to the ratio and incubate at room temperature, protected from light, for 90 min. Commonly used secondary antibody ratios: goat anti-rabbit IgG H&L (Alexa Fluor®647) (1:500, A-31573, Invitrogen), goat anti-chicken IgY H&L (Alexa Fluor® 555) (1:500, A-21437, Invitrogen), goat anti-mouse IgG H&L (Alexa Fluor® 488) (1:500, A-11001, Invitrogen); (8) Wash three times with 0.01M PBS at room temperature, away from light, for 10 min each time; (9) Add anti-fluorescence quenching mounting solution containing DAPI to mount the slides; (10) Observe and take pictures under a fluorescence microscope; (11) Use ImageJ software to measure the fluorescence intensity of the images.

[0052] 5. Experimental results

[0053] The results showed that Achyranthes bidentata peptide combined with human umbilical cord mesenchymal stem cell exosomes can significantly alleviate the motor dysfunction of Parkinson's model mice. For example, in the pole climbing experiment, the time taken by treated mice to complete the pole climbing was significantly lower than that of the MPTP-treated group ( Figure 3 B). In addition, this treatment can significantly improve the olfactory function of Parkinson's model mice, as shown by the significantly shorter time it takes for the treated group mice to find hidden food ( Figure 3C). The above behavioral experimental results showed that the combined use of drugs had a more prominent effect compared with the treatment with Achyranthes bidentata peptide or exosomes alone. The results of tissue morphological analysis showed that the expression of tyrosine hydroxylase (TH) in the substantia nigra of PD model mice was significantly reduced, while the use of Achyranthes bidentata peptide combined with human umbilical cord mesenchymal stem cell exosomes could significantly increase the expression of TH, indicating that the number of dopaminergic neurons was increased ( Figure 4 A, B). In addition, this treatment can also inhibit the activation of microglia and astrocytes in the midbrain, as shown by a decrease in the number of Iba-1 and GFAP-positive cells ( Figure 5 A, B; Figure 6 A, B). Histomorphological analysis of the olfactory bulb revealed that the combination of Achyranthes bidentata peptide and human umbilical cord mesenchymal stem cell exosomes significantly enhanced the expression of phosphorylated S6 ribosomal protein (Phospho-S6Ribosomal Protein), indicating that the activity of olfactory bulb neurons was improved ( Figure 7 At the same time, the activation of microglia in the lateral and central regions of the olfactory bulb was inhibited, as evidenced by the decrease in Iba-1 positive cells after treatment ( Figure 8 In addition, the activation of astrocytes in the lateral region of the olfactory bulb was also inhibited, as evidenced by the decrease in GFAP-positive cells after treatment ( Fig. 9 ). The above morphological experimental results show that compared with the treatment of Achyranthes bidentata peptide or exosomes alone, the combined treatment has a more prominent effect.

[0054] Although the above embodiments have described the present invention in detail, it is not limited to the above implementation modes. Practitioners in this field may make other modifications and improvements based on the inspiration of the present invention, which all fall within the protection scope of the present invention.

Claims

1. Application of Achyranthes bidentata polypeptide combined with human umbilical cord mesenchymal stem cell exosomes in the preparation of drugs for preventing and / or treating Parkinson's disease.

2. The use according to claim 1, characterized in that: The drug includes 1~2×10 8 / kg human umbilical cord mesenchymal stem cell exosomes and 50~100 μg / kg Achyranthes bidentata peptides.

3. The use according to claim 1 or 2, characterized in that: The Achyranthes bidentata polypeptide is prepared by the following method: Cutting the dried root of Achyranthes bidentata into pieces, mixing the cut dried root of Achyranthes bidentata with water and then steaming to obtain a mixed liquid; Adding ammonium sulfate to the mixed solution for precipitation, centrifuging, and freeze-drying to obtain a crude extract of Achyranthes bidentata polypeptide; The crude extract of Achyranthes bidentata polypeptide is dissolved in water to prepare a solution, and then the solution is purified by high performance liquid chromatography to obtain the Achyranthes bidentata polypeptide.

4. The use according to claim 3, characterized in that: The cooking temperature is 80-85°C and the cooking time is 40-60min.

5. The use according to claim 3, characterized in that: The conditions of the HPLC are as follows: C18 reverse phase column, mobile phase is water / acetonitrile containing 0.1% trifluoroacetic acid, the volume ratio of water to acetonitrile is 4:1, flow rate is 1.0 mL / min, and UV detector is at a wavelength of 220 nm.

6. The use according to claim 1 or 2, characterized in that: The human umbilical cord mesenchymal stem cell exosomes are prepared by the following method: The primary human umbilical cord mesenchymal stem cells were subcultured in a serum-containing medium. When the density of the third generation human umbilical cord mesenchymal stem cells reached 80%, the medium was changed to a serum-free medium and cultured for 48 hours, and the supernatant was collected. The supernatant was then centrifuged at 400-500 g to remove cells and cell debris, and then filtered through a 0.22 μm filter; The filtered supernatant is treated with an exosome separation kit, and the umbilical cord mesenchymal stem cell exosomes are obtained by separation and purification through a specific buffer and centrifugation step.

7. The use according to claim 1, characterized in that: The application includes any of the following: Improve motor dysfunction and olfactory perception; Increase the number of dopaminergic neurons in the brain; Inhibit the abnormal activation of microglia and astrocytes in the brain; Enhances the activity of olfactory bulb neurons.

8. The use according to claim 1, characterized in that: The drug is administered through the nasal cavity.

9. The use according to claim 1, characterized in that: The medicine is a solution.

10. The use according to claim 1, characterized in that: The drug also includes pharmaceutically acceptable excipients.

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