Application of ganoderma lucidum extract in aspect of improving influence of intestinal flora on neurodegenerative diseases

Abstract

The invention discloses application of a ganoderma lucidum extract in the aspect of regulating intestinal flora and improving neurodegenerative diseases, and the ganoderma lucidum extract composed of glucose has influence on the richness and uniformity of the intestinal flora of the neurodegenerative disease Parkinson's disease. The dominant bacteria of the intestinal flora after the administration of the lucid ganoderma extract belong to the genus Lactobacillus, the genus Enterobacter and the anti-inflammatory related Muribacaceae, and the dominant bacteria of the intestinal flora after the administration of the lucid ganoderma extract belong to the genus Lactobacillus, the genus Enterobacter and the anti-inflammatory related Muribacaceae. The weight of the mouse is increased, the behavioral ability is improved, and the lucid ganoderma extract has influence on the composition diversity and community difference of intestinal flora of the neurodegenerative disease Parkinson's disease and the abundance of dominant bacteria.

Description

Technical Field

[0001] The invention relates to the preparation of ganoderma lucidum extract and its application in regulating intestinal flora to improve neurodegenerative diseases. Background Art

[0002] Neurodegenerative diseases are characterized by varying degrees of cognitive impairment and movement disorders as their main clinical manifestations, and their prevalence has gradually increased in recent years. Currently, the treatment of neurodegenerative diseases is mainly based on drugs, and there is no treatment that can reverse the outcome of the disease. Parkinson's disease is a degenerative disease of the central nervous system that occurs frequently in middle-aged and elderly people, and is mainly manifested by motor dysfunction and non-motor symptoms. In recent years, there have been reports that Parkinson's disease can be affected by improving intestinal flora.

[0003] Traditional Chinese medicine has been widely and deeply studied in the treatment of neurological diseases. Ganoderma lucidum belongs to the genus Ganoderma of the family Ganodermaceae. It is a polyporous fungus that is widely distributed, mainly in tropical and subtropical regions. The main application part of Ganoderma lucidum listed in the Pharmacopoeia of the People's Republic of China is the fruiting body, and one of the main active and nutritious components in the fruiting body of Ganoderma lucidum is polysaccharides. Natural polysaccharides can be extracted from animals, plants and traditional Chinese medicine. They are an important source of energy for the intestinal microbiome, promote the proliferation of probiotics and regulate the balance of intestinal microecology. Previous studies have shown that Ganoderma lucidum polysaccharides can regulate intestinal flora disorders to achieve anti-inflammatory effects. The present invention aims to study whether Ganoderma lucidum polysaccharides can delay neurodegenerative diseases by maintaining intestinal balance. Summary of the invention

[0004] The purpose of the present invention is to provide an application of Ganoderma lucidum extract in improving neurodegenerative diseases in terms of intestinal flora. Specifically, the present invention analyzes the intestinal flora of mice with the neurodegenerative disease Parkinson's disease (PD) after being given Ganoderma lucidum extract, and explores the improvement effect of Ganoderma lucidum extract on neurodegenerative diseases based on the changes in intestinal flora.

[0005] The ganoderma lucidum extract of the present invention is prepared according to a method comprising the following steps:

[0006] Grinding and sieving process: After washing the Ganoderma lucidum fruiting bodies, dry them at 55-65℃ until constant weight, grind them in a grinder, and sieve them to obtain Ganoderma lucidum powder.

[0007] Degreasing process: add 95% ethanol, soak and let stand overnight, filter and place in oven.

[0008] Water extraction and alcohol precipitation process: weigh a certain amount of powder, add ultrapure water to soak overnight, heat reflux extraction twice for 2-4 hours each time, collect the precipitate by centrifugation, combine the extracts twice and concentrate under reduced pressure, add 95% ethanol to make the ethanol concentration reach 60%, let it stand overnight, filter, add ethanol to make its concentration reach 80% and let it stand overnight, and freeze-dry the precipitate after centrifugation.

[0009] Separation and purification process: After the above powder is dissolved in phosphate buffered saline, it is purified by Q-FF agarose gel, eluted with 0.2NaCl, the eluate is collected, dialyzed and freeze-dried to obtain a crude polysaccharide powder of Ganoderma lucidum. The crude polysaccharide powder of Ganoderma lucidum is further purified by dextran G-100, eluted with water, the eluate is collected, and freeze-dried to obtain a Ganoderma lucidum extract.

[0010] Preferably, in step (1), the oven drying temperature of the washed Ganoderma lucidum fruiting bodies is 55-66°C, more preferably 65°C.

[0011] Preferably, the heating extraction temperature in step (3) is 60-100°C, more preferably 80-90°C.

[0012] Preferably, the heating reflux extraction time in step (3) is 2-4 hours, preferably 3 hours at a time.

[0013] Preferably, the ratio of ultrapure water added in step (3) is 1:10-30 times, and more preferably 1:20 times

[0014] Preferably, in step (2), the volume ratio of adding 95% ethanol to liquid is 1:4.

[0015] The preferred extraction process comprises the following specific steps:

[0016] Grinding and sieving process: After washing the Ganoderma lucidum fruiting bodies, place them in a 65℃ constant temperature drying oven until constant weight is reached, grind them with a grinder, and pass through an 80-mesh sieve to obtain Ganoderma lucidum powder.

[0017] Add 95% ethanol at a material-liquid ratio of 1:4 (g:ml), soak overnight to remove lipid substances, filter and place the precipitate in a constant temperature drying oven to obtain Ganoderma lucidum powder

[0018] Weigh a certain amount of powder, add ultrapure water, heat and reflux extraction twice for 3 hours each time, the heating temperature is 80 degrees Celsius, filter and collect the precipitate, combine the two extracts and concentrate under reduced pressure to 50 ml, add 95% ethanol to make the ethanol concentration reach 60%, let it stand overnight, filter, add ethanol to make its concentration reach 80% and let it stand overnight, and freeze-dry the precipitate after centrifugation to obtain Ganoderma lucidum powder.

[0019] Separation and purification process: After the above powder is dissolved in phosphate buffered saline, it is purified by Q-FF agarose gel, eluted with 0.2NaCl, the eluate is collected, dialyzed and freeze-dried to obtain a crude polysaccharide powder of Ganoderma lucidum. The crude polysaccharide powder of Ganoderma lucidum is further purified by dextran G-100, eluted with water, the eluate is collected, and freeze-dried to obtain a Ganoderma lucidum extract.

[0020] The specific steps of preparing the ganoderma lucidum extract of the present invention are as follows:

[0021] Take a clean chromatographic bottle, weigh an appropriate amount of sample, add 1ml of 2M trifluoroacetic acid solution, and heat at 121℃ for 2 hours. Pass nitrogen and blow dry. Add 99.99% methanol to wash, blow dry again, and repeat the methanol washing 2-3 times. Add sterile water to dissolve and transfer to the chromatographic bottle for testing. The chromatographic system used is the Thermo ICS 5000+ ion chromatography system (ICS 5000+, ThermoFisher Scientific, USA), and the monosaccharide components are analyzed and detected using an electrochemical detector. A DionexTMCarboPacTMPA20 (150*3.0mm, 10μm) liquid chromatography column is used; the injection volume is 5μl. Mobile phase A (H2O), mobile phase B (0.1 M NaOH), mobile phase C (0.1 M NaOH, 0.2 M NaAc), flow rate 0.5 ml / min; column temperature was 30°C; elution gradient: 0 min A phase / B phase / C phase (95:5:0, V / V), 26 min A phase / B phase / C phase (85:5:10, V / V), 42 min A phase / B phase / C phase (85:5:10, V / V), 42.1 min A phase / B phase / C phase (60:0:40, V / V), 52 min A phase / B phase / C phase (60:40:0, V / V), 52.1 min A phase / B phase / C phase (95:5:0, V / V), 60 min A phase / B phase / C phase (95:5:0, V / V).

[0022] First, the present invention provides the use of a Ganoderma lucidum extract in the preparation of a drug for regulating intestinal flora. Preferably, the drug is used to regulate intestinal flora imbalance caused by neurodegenerative diseases.

[0023] Furthermore, the neurodegenerative disease is Parkinson's disease.

[0024] Preferably, the application is any one of the following 1)-4):

[0025] 1) Beta diversity analysis of different communities of intestinal flora

[0026] 2) Regulate the Alpha Diversity of Intestinal Microbiota

[0027] The intestinal flora diversity is mainly analyzed by Chao1, Shannon, and Simpson

[0028] 3) Changes in the abundance of key dominant bacteria that regulate intestinal flora

[0029] The key dominant bacteria of the intestinal flora are Lactobacillus, Enterobacter

[0030] (Enterorhabdus), and Muribaculaceae, which is associated with anti-inflammatory

[0031] 4) Regulate intestinal flora function

[0032] The present invention also provides the use of the ganoderma lucidum extract in preparing a product having any one of the following functions 1)-4):

[0033] 1) Beta diversity analysis of different communities of intestinal flora

[0034] 2) Regulate the Alpha Diversity of Intestinal Microbiota

[0035] The intestinal flora diversity is mainly analyzed by Chao1, Shannon, and Simpson

[0036] 3) Changes in the abundance of key dominant bacteria that regulate intestinal flora

[0037] The key dominant bacteria of the intestinal flora are Lactobacillus, Enterobacter

[0038] (Enterorhabdus), and Muribaculaceae, which is associated with anti-inflammatory

[0039] 4) Regulate intestinal flora function

[0040] Preferably, the present invention also provides the use of Ganoderma lucidum extract in the preparation of a drug for improving Parkinson's disease.

[0041] Preferably, the application is any one of the following 1)-2):

[0042] 1) Gaining weight

[0043] 2) Improve behavioral abilities

[0044] The fatigue rotarod test for improving behavioral ability is any one of the following a)-b):

[0045] a) Reduce the number of drops

[0046] b) Increase the maximum stay time

[0047] The present invention also provides the use of the ganoderma lucidum extract in preparing a product having any of the following functions 1)-2) 1) increasing body weight

[0048] 2) Improve behavioral abilities

[0049] The fatigue rotarod test for improving behavioral ability is any one of the following a)-b):

[0050] a) Reduce the number of drops

[0051] b) Increase the maximum stay time

[0052] The preliminary analysis of the intestinal flora sequencing results of the present invention shows that the administration of Ganoderma lucidum extract has an effect on the intestinal flora structure of mice with neurodegenerative diseases, and there are also differences between the groups, indicating that the changes in the intestinal flora between the neurodegenerative disease group and the Ganoderma lucidum extract administration group are also different. Therefore, it is speculated that the protective effect of Ganoderma lucidum extract on neurodegenerative diseases may be related to its influence on the structure of the intestinal flora community.

[0053] In the study of the neuroprotective effect of Ganoderma lucidum extract on neurodegenerative diseases, the administration of Ganoderma lucidum extract increased the weight of mice, reduced the number of falls on the fatigue rod, and increased the maximum stay time of mice on the axis. Ganoderma lucidum extract administration has an effect on the species composition, diversity of flora composition, abundance of dominant bacteria and functionality of the intestinal flora of mice with neurodegenerative diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 This is a composition diagram of Ganoderma lucidum extract.

[0055] Figure 2 Beta diversity analysis diagram of different communities of intestinal flora regulating the intestine.

[0056] Figure 3 Analyze Chao1 graph for Alpha.

[0057] Figure 4 Analyze the Shannon plot for Alpha.

[0058] Figure 5 Analyze the Simpson plot for Alpha.

[0059] Figure 6 This is a door level analysis diagram.

[0060] Figure 7 This is an analysis chart of key dominant bacteria at the genus level.

[0061] Figure 8 Heat map of community composition at the genus level combined with cluster analysis. DETAILED DESCRIPTION

[0062] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0063] All materials and reagents in the following examples were obtained from commercial sources.

[0064] Example 1: Regulatory effect of Ganoderma lucidum extract on intestinal flora in neurodegenerative diseases

[0065] 1. Preparation of Ganoderma lucidum extract:

[0066] 1. After washing the fruiting bodies of Ganoderma lucidum (Ganoderma lucidum with flat cap from Changbai Mountain, Jilin Province), put them in a 65℃ constant temperature drying oven until constant weight, crush them with a pulverizer, and pass through an 80-mesh sieve to obtain Ganoderma lucidum powder. Add anhydrous ethanol at a solid-liquid ratio of 1:4 (g:ml), soak overnight, remove lipid substances, filter and place the precipitate in a constant temperature drying oven to obtain Ganoderma lucidum powder, weigh a certain amount of powder, add ultrapure water, heat and reflux extraction 2 times for 3 hours each time, the heating temperature is 80 degrees Celsius, filter and collect the precipitate, combine the extracts twice and concentrate under reduced pressure to 50ml, add 95% ethanol at a ratio of 1:4 (ml:ml) to an ethanol concentration of 60%, let stand overnight, filter, add ethanol to a concentration of 80% and let stand overnight, freeze-dry the precipitate after centrifugation to obtain Ganoderma lucidum powder. The above powder was dissolved in phosphate buffer, purified by Q-FF agarose gel, eluted with 0.2NaCl, the eluate was collected, dialyzed and freeze-dried to obtain a crude polysaccharide powder of Ganoderma lucidum. The crude polysaccharide powder of Ganoderma lucidum was further purified by dextran G-100, eluted with water, the eluate was collected, and freeze-dried to obtain a Ganoderma lucidum extract.

[0067] 2. Composition of Ganoderma Lucidum Extract:

[0068] Take a clean chromatographic bottle, weigh an appropriate amount of sample, add 1 ml of 2M trifluoroacetic acid solution, and heat at 121°C for 2 hours. Pass nitrogen and blow dry. Add 99.99% methanol to wash, blow dry again, and repeat the methanol washing 2-3 times. Add sterile water to dissolve and transfer to the chromatographic bottle for testing. The chromatographic system used is the Thermo ICS 5000+ ion chromatography system (ICS 5000+, ThermoFisher Scientific, USA), and the monosaccharide components are analyzed and detected using an electrochemical detector.

[0069] A Dionex™ CarboPac™ PA20 (150*3.0 mm, 10 μm) liquid chromatography column was used; the injection volume was 5 μl. Mobile phase A (H2O), mobile phase B (0.1M NaOH), mobile phase C (0.1M NaOH, 0.2M NaAc), flow rate 0.5ml / min; column temperature 30℃; elution gradient: 0min A phase / B phase / C phase (95:5:0, V / V), 26min A phase / B phase / C phase (85:5:10, V / V), 42min A phase / B phase / C phase (85:5:10, V / V), 42.1min A phase / B phase / C phase (60:0:40, V / V), 52min A phase / B phase / C phase (60:40:0, V / V), 52.1min A phase / B phase / C phase (95:5:0, V / V), 60min A phase / B phase / C phase (95:5:0, V / V). Chromatographic data were processed using Chromeleon software. The figure below shows the standard sample ion chromatogram and the sample ion chromatogram. The horizontal axis is the retention time of the detection (Time, min), and the vertical axis is the response value of the ion detection (Response, nC).

[0070] 3. Detection of intestinal flora in mice with neurodegenerative diseases

[0071] 1. Grouping of experimental animals and establishment of neurodegenerative disease models

[0072] After one week of adaptive feeding, 30 C57BL / 6 mice were divided into three groups. The mice in the Ganoderma lucidum extract administration group were gavaged with a dose of 60 mg / kg, and the mice in the neurodegenerative disease group were given MPTP to establish a Parkinson's injury model. On the eighth day, the drug administration group and the model group were injected with MPTP at a dose of 30 mg / kg, and the control group was given the same dose of normal saline, and the weight of the mice was recorded.

[0073] 2. Collection of Mouse Samples

[0074] On the 15th day after administration, fresh mouse feces (wet weight 1 g) were collected and stored at -80°C for testing.

[0075] 3. Intestinal flora detection

[0076] (1) DNA extraction from stool samples

[0077] The DNA in mouse fecal samples was quantified by Nanodrop, and the quality of DNA extraction was checked by 1.2% agarose gel electrophoresis.

[0078] (2) PCR amplification of target fragment

[0079] PCR amplification uses Pfu high-fidelity DNA polymerase from Quanshijin Company, and strictly controls the number of amplification cycles to keep the number of cycles as low as possible while ensuring that the amplification conditions of the same batch of samples are consistent. At the same time, a negative control is set up to detect microbial contamination such as the environment and reagents. Any sample group with bands in the negative control amplification cannot be used for subsequent experiments.

[0080] (3) Purification and recovery of amplified products by magnetic beads

[0081] Add 0.8 times the volume of magnetic beads (Vazyme VAHTSTM DNA Clean Beads) to 25μl PCR product, shake to fully suspend, adsorb on a magnetic rack for 5 minutes, and carefully aspirate the supernatant with a pipette; add 20μl 0.8 times magnetic bead washing solution, shake to fully suspend, place on a magnetic rack for adsorption for 5 minutes, and carefully aspirate the supernatant; add 200μl 80% ethanol, place on the magnetic rack in reverse to adsorb the magnetic beads to the other side of the PCR tube, and aspirate the supernatant after sufficient adsorption; let it stand at room temperature for 5 minutes until the alcohol evaporates completely and cracks appear on the magnetic beads; add 25μl Elution Buffer for elution; place the PCR tube on the adsorption rack for 5 minutes, fully adsorb, and remove the supernatant to a clean 1.5ml centrifuge tube for storage.

[0082] (4) Fluorescence quantification of amplified products

[0083] The PCR amplification products were subjected to fluorescence quantification, the fluorescence reagent was Quant-iT PicoGreen dsDNAAssay Kit, and the quantitative instrument was Microplate reader (BioTek, FLx800). According to the fluorescence quantification results and the sequencing amount requirements of each sample, the samples were mixed in corresponding proportions.

[0084] (5) Sequencing library preparation

[0085] The sequencing library was prepared using Illumina's TruSeq Nano DNA LT Library Prep Kit.

[0086] (6) Perform high-throughput sequencing on the machine

[0087] Before sequencing, the library needs to be quality checked on the Agilent Bioanalyzer using the Agilent High Sensitivity DNA Kit. A qualified library has only a single peak and no connectors. Afterwards, the library is quantified on the Promega QuantiFluor fluorescence quantitative system using the Quant-iT PicoGreen dsDNA Assay Kit, and the concentration of a qualified library should be above 2 nM. After gradient dilution of each qualified sequencing library (Index sequence cannot be repeated), mix them in the corresponding proportion according to the required sequencing amount, and denature them into single strands with NaOH for sequencing; use the MiSeq sequencer for double-end sequencing, and the corresponding reagent is the MiSeq Reagent Kit V3 (600 cycles). The optimal sequencing length of the target fragment is 200 to 450 bp.

[0088] (7) Microbial diversity analysis, principal component analysis and sample cluster analysis

[0089] Sequence denoising or OTU clustering was performed according to the QIIME2 dada2 analysis process or the Vsearch software analysis process. The specific composition of each sample (group) at different species taxonomic levels was displayed to understand the overall profile. According to the distribution of ASV / OTU in different samples, the Alpha diversity level of each sample was evaluated, the distance matrix of each sample was calculated, and the beta diversity differences and significance of differences between different samples, as well as the differences in species abundance composition between different samples were measured through a variety of unsupervised sorting and clustering methods combined with corresponding statistical test methods.

[0090] 4. Fatigue Rotarod Test

[0091] The mice were placed on the exercise rod at the same time, and a three-minute adaptation experiment was first conducted to adapt the mice to the roller, with a rotation speed of 20r / min. After the adaptation, the mice were placed on the exercise rod again, with 5 minutes as a group, and each mouse was tested 5 times to take the average value to eliminate the existence of contingency. After each test, a 1-minute rest was performed. The number of times each mouse fell and the maximum stay time on the roller were carefully recorded, and finally sorted by grouping.

[0092] 5. Composition of Ganoderma Lucidum Extract

[0093] The present invention analyzes the Ganoderma lucidum extract through an electrochemical detector and finds that the Ganoderma lucidum extract is mainly composed of polysaccharides ( Figure 1 ).

[0094] VI. Results of intestinal flora detection in mice with neurodegenerative diseases

[0095] The preliminary analysis of the intestinal flora sequencing results of the present invention shows that the administration of Ganoderma lucidum extract has an effect on the intestinal flora structure of mice with neurodegenerative diseases, and there are also differences between the groups, indicating that the changes in the intestinal flora between the neurodegenerative disease group and the Ganoderma lucidum extract administration group are also different. Therefore, it is speculated that the protective effect of Ganoderma lucidum extract on neurodegenerative diseases may be related to its influence on the structure of the intestinal flora community.

[0096] 1. Beta diversity analysis of different communities of intestinal flora

[0097] PCA analysis revealed that there were significant differences in the distribution of sample communities in each group. The distance between the control group and the model group mouse samples was relatively large, and there was a significant difference in the composition of the microbial community. After the administration of Ganoderma lucidum extract, the sample community tended to be normal, indicating that Ganoderma lucidum extract can improve the intestinal community. However, there was also a significant difference in the composition of the microbiota between the control group and the administration group ( Figure 2 ).

[0098] 2. Analysis of Alpha Diversity of Intestinal Flora

[0099] Chao1, Shannon and Simpson were used to analyze the species diversity of the microbiota in the three groups of samples. It was found that the species diversity in the samples after drug administration was greater than that in the model group, and the microbiota diversity in the model group was lower than that in the control group. This result suggests that compared with the model group, the intestinal microbiota structure of Parkinson's disease mice after administration of Ganoderma lucidum extract tends to be healthier ( Figure 3-5 ).

[0100] 3. Changes in abundance of key dominant bacterial communities

[0101] In the analysis of different groups in the structure of the mouse intestinal microbial flora, based on the phylum level analysis, the largest proportion of flora are all concentrated in the Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria ( Figure 6 At the genus level, the top 15 bacterial genera with the most significant differences among the groups were screened ( Figure 7 ). By comparing the proportion of bacterial flora in the model group and the drug-treated group, the most significant changes in abundance were in Lactobacillus, Enterobacter (Enterorhabdus), and Muribaculaceae, which is related to anti-inflammatory. Literature reports that changes in the abundance of these genera are correlated with Parkinson's disease.

[0102] 4. Changes in bacterial flora functions

[0103] Through heat map and abundance calculation, the results showed that the intestinal flora functions were different before and after the administration of Ganoderma lucidum extract ( Figure 8 ).

[0104] 7. Research results on the effect of Ganoderma lucidum extract on improving Parkinson's neurodegenerative disease

[0105] 1. Weight changes

[0106] As shown in Table 1, the weight changes of mice in the blank group showed a normal growth level, while the weight of mice in the model group showed a slow growth trend after intraperitoneal injection of MPTP, and the weight growth of mice in the Ganoderma lucidum extract-administered group gradually tended to that of normal mice.

[0107] Table 1 Body weight changes

[0108]

[0109] Note: Compared with the blank group *P<0.05, **P<0.01; compared with the model group #P<0.05, ##P<0.01, ###P<0.001

[0110] Behavioral influence

[0111] 2. Fatigue Rotarod Test

[0112] 2.1 Drop times

[0113] The results showed that the number of falls in the blank group was the least, the Ganoderma lucidum extract group was the second, and the model group had the most falls. In other words, compared with the blank group, the number of falls in the model group increased significantly (P<0.05), and the number of falls in the Ganoderma lucidum extract group was significantly reduced and approached the normal level. Therefore, the administration of Ganoderma lucidum extract can effectively prevent and treat movement disorders, as shown in Table 2.

[0114] Table 2 Number of drops

[0115]

[0116] Note: *P<0.05, **P<0.01

[0117] 2.2 Maximum residence time

[0118] The results showed that the blank group had the longest residence time, followed by the drug-treated group, and the model group had the shortest residence time, indicating that the residence time of the model group was significantly decreased compared with the blank group (P<0.05). Ganoderma lucidum extract can effectively prevent and treat balance disorders, as shown in Table 3.

[0119] Table 3 Maximum residence time

[0120]

[0121] Note: *P<0.05, **P<0.01.

Claims

1. Application of Ganoderma lucidum extract in the preparation of drugs for regulating intestinal flora The ganoderma lucidum extract is prepared according to a method comprising the following steps: 1) Grinding and sieving process: After washing the Ganoderma lucidum fruiting bodies, dry them at 55-65° C. to constant weight, grind them in a grinder, and sieve them to obtain Ganoderma lucidum powder; 2) Degreasing process: add 95% ethanol, soak and let stand overnight, filter and place in oven; 3) Water extraction and alcohol precipitation process: weigh a certain amount of powder, add ultrapure water to soak overnight, heat reflux extraction twice for 2-4 hours each time, collect the precipitate by centrifugation, combine the extracts twice and concentrate under reduced pressure, add 95% ethanol to an ethanol concentration of 60%, let stand overnight, filter, add ethanol to a concentration of 80% and let stand overnight, and freeze-dry the precipitate after centrifugation; 4) Separation and purification step: dissolving the above powder with phosphate buffered saline solution, purifying with Q-FF agarose gel, eluting with 0.2 NaCl, collecting the eluate, dialyzing and freeze-drying to obtain Ganoderma lucidum polysaccharide powder, further purifying the Ganoderma lucidum crude polysaccharide powder with dextran G-100, eluting with water, collecting the eluate, and freeze-drying to obtain Ganoderma lucidum extract; 5) The ganoderma lucidum extract is a polysaccharide mainly composed of glucose.

2. The use according to claim 1, characterized in that: The drug is used to regulate the imbalance of intestinal flora caused by the neurodegenerative disease Parkinson's disease.

3. The use according to claim 1 or 2, characterized in that: The application is any one of the following 1)-4): 1) Beta diversity analysis of different communities regulating intestinal flora; 2) Regulate the Alpha diversity of intestinal flora; The intestinal flora diversity is mainly analyzed by Chao1, Shannon, and Simpson 3) Changes in the abundance of key dominant bacteria that regulate intestinal flora; 4) Regulate the function of intestinal flora.

4. The use according to claim 3, characterized in that: The dominant bacteria are Lactobacillus, Enterobacter (Enterorhabdus), and Muribaculaceae associated with anti-inflammation.

5. Application of Ganoderma lucidum extract in the preparation of products having any of the following functions 1)-4): 1) Beta diversity analysis of different communities of intestinal flora; 2) Regulate the Alpha diversity of intestinal flora; The intestinal flora diversity is mainly analyzed by Chao1, Shannon, and Simpson 3) Changes in the abundance of key dominant bacteria that regulate intestinal flora; 4) Regulate the function of intestinal flora.

6. The use according to claim 5, characterized in that: The dominant bacteria are Lactobacillus, Enterobacter (Enterorhabdus), and Muribaculaceae associated with anti-inflammation.

7. Application of Ganoderma lucidum extract in the preparation of medicines for improving the neurodegenerative disease Parkinson's disease.

8. The use according to claim 7, characterized in that: The application is any one of the following 1)-2) 1) Increase weight; 2) Improve behavioral abilities.

9. The use according to claim 8, characterized in that: The fatigue rotarod test for improving behavioral ability is any one of the following ab: a) Reduce the number of drops; b) Increase the maximum dwell time.

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