Application of dendrobium polysaccharide in preparation of medicine for preventing and / or treating neuroinflammation caused by alcohol and dendrobium polysaccharide liqueur

Abstract

The invention provides application of dendrobium polysaccharide in preparation of a medicine for preventing and / or treating neuroinflammation caused by alcohol and dendrobium polysaccharide liqueur, and belongs to the field of biological medicine. According to the invention, the dendrobium polysaccharide is prepared into the liqueur, and the liqueur is applied to a mouse model exposed to alcohol for a long time, so that the dendrobium polysaccharide liqueur is found to be capable of improving neurological dysfunction such as cognitive dysfunction, motor coordination ability decline and anxiety behavior caused by long-term drinking; the neuroinflammation of long-term drinking mice can be intervened. It is indicated that the dendrobium polysaccharide has a remarkable intervention effect on the neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease and the like, caused by excessive drinking of alcohol. A new way is provided for the development and utilization of dendrobium resources, and the modernization of traditional Chinese medicines and the development of the large health industry are promoted.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular relates to the use of dendrobium polysaccharide in preparing a medicine for preventing and / or treating alcohol-induced neuroinflammation and a dendrobium polysaccharide liquor. Background Art

[0002] Chronic alcohol abuse is a global health problem. Alcohol has multiple damaging effects on the nervous system, with neuroinflammation being a key pathological mechanism. Alcohol-induced neuroinflammation can lead to a range of neurological disorders, including neuronal damage, neurotransmitter imbalance, and cognitive impairment, such as alcoholic encephalopathy and neurodegenerative diseases.

[0003] Lujiu (Lujiu) refers to a beverage made with fermented wine, distilled wine, or edible alcohol as its base, blended with edible or medicinal ingredients. It possesses a unique flavor and certain health benefits. Dendrobium officinale is a traditional and valuable Chinese medicinal herb rich in various bioactive components, among which polysaccharides exhibit antioxidant, anti-tumor, and immunomodulatory activities. However, there are no reports on the interventional effects of dendrobium polysaccharide Lujiu on neuroinflammation induced by long-term alcohol exposure.

[0004] Currently, clinical treatments for alcoholic neuroinflammation are limited and have significant side effects. Therefore, developing safe and effective interventions is of great significance. Summary of the Invention

[0005] In light of this, the present invention aims to provide the use of dendrobium polysaccharides in the preparation of a medicament for preventing and / or treating alcohol-induced neuroinflammation, as well as a dendrobium polysaccharide wine. This invention prepares dendrobium polysaccharides into a wine and applies it to a mouse model exposed to long-term alcohol exposure. The wine demonstrates a significant interventional effect on neuroinflammation. Furthermore, the wine offers advantages such as ease of administration, pleasant taste, and high safety, making it readily accepted by consumers and promising broad market prospects.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides the use of dendrobium polysaccharide in preparing a medicament for preventing and / or treating alcohol-induced neuroinflammation, wherein the alcohol-induced neuroinflammation is an alcoholic neurological disease caused by excessive drinking.

[0008] Preferably, the alcoholic neurological diseases caused by excessive drinking include alcoholic encephalopathy and neurodegenerative diseases caused by excessive drinking.

[0009] Preferably, the medicine is in the form of tablets, powders, granules, capsules, pills, aqueous solutions, alcoholic solutions or syrups.

[0010] The present invention provides a dendrobium polysaccharide wine for preventing and / or treating alcohol-induced neuritis, comprising a base wine with an alcohol concentration of 30-50% and dendrobium polysaccharide; the mass volume ratio of the dendrobium polysaccharide to the base wine is 1-10g:1L.

[0011] Preferably, 1L of base wine also includes 50-200g of flavoring agent.

[0012] Preferably, the flavoring agent is honey.

[0013] The present invention also provides a method for preparing the dendrobium polysaccharide wine, which comprises dissolving the dendrobium polysaccharide in base wine, homogenizing and aging the wine to obtain the dendrobium polysaccharide wine.

[0014] Preferably, the homogenization pressure is 20-30 MPa, the homogenization times are 2-3 times, and the homogenization time is 3-5 min / time.

[0015] Preferably, the aging temperature is 15-25° C., the aging relative humidity is 60%-75%, and the aging time is 0.5-1.5 months.

[0016] Preferably, the step of adding flavoring agent is included before homogenization, and the step of filtering is included after aging.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] (1) The present invention discovered for the first time that dendrobium polysaccharides and dendrobium polysaccharide wine have a significant intervention effect on neuroinflammation caused by long-term exposure to alcohol. Specifically, dendrobium polysaccharide wine can improve neurological dysfunction such as cognitive dysfunction, decreased motor coordination ability, and anxiety-like behavior caused by long-term drinking; it can intervene in neuroinflammation in long-term drinking mice by changing their neurobehavioral, inflammatory factors, neurotransmitters, hippocampal structure, and oxidative stress and other physiological and biochemical indicators. This shows that dendrobium polysaccharides have a significant intervention effect on neurodegenerative diseases caused by excessive alcohol consumption, including Alzheimer's disease and Parkinson's disease. Compared with traditional drug treatments, dendrobium polysaccharide wine has the advantages of being easy to take, having a good taste, and being highly safe. It is easily accepted by consumers and has broad market application prospects.

[0019] (2) The present invention provides a new approach for the development and utilization of Dendrobium resources, which will help promote the modernization of traditional Chinese medicine and the development of the health industry. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1The results of the behavioral effects of DOP on long-term alcohol-drinking mice are shown in Figure 2. Among them, a is the result of the Y maze test; b is the result of the mining test; c is the result of the fatigue rotarod test; the BLK group is the blank control group; the E_ALC group is the model group; the LU group is the positive drug group; and DOP is the Dendrobium polysaccharide wine group.

[0021] Figure 2 The results of DOP's effects on inflammatory factors in the brain tissue of mice exposed to long-term alcohol consumption are shown in Figure 2. a) shows the effect on IL-6 content; b) shows the effect on IL-1β content; and c) shows the effect on TNF-α content.

[0022] Figure 3 The figure shows the effect of DOP on neurotransmitters in the hippocampus of long-term alcohol-drinking mice; a shows the effect on 5-hydroxytryptamine (5-HT) content; b shows the effect on dopamine (DA) content; c shows the effect on γ-aminobutyric acid (GABA) content; and d shows the effect on glutamate (Glu) content.

[0023] Figure 4 The results of the effects of DOP on the CA3 structure in the hippocampus of mice with long-term alcohol consumption are shown in Figure 2. a is the BLK group; b is the E_ALC group; c is the LU group; d is the DOP group;

[0024] Figure 5 The figure shows the effect of DOP on oxidative stress indicators in the brain tissue of mice with long-term alcohol consumption; a is the effect on the content of reduced glutathione (GSH); b is the effect on the content of malondialdehyde (MDA); and c is the effect on the content of superoxide dismutase (T-SOD). DETAILED DESCRIPTION

[0025] The present invention provides the use of dendrobium polysaccharide in preparing a medicament for preventing and / or treating alcohol-induced neuroinflammation, wherein the alcohol-induced neuroinflammation is an alcoholic neurological disease caused by excessive drinking.

[0026] In the present invention, the alcoholic neurological diseases caused by excessive drinking include alcoholic encephalopathy and neurodegenerative diseases caused by excessive drinking, and the neurodegenerative diseases caused by excessive drinking include Alzheimer's disease and Parkinson's disease.

[0027] In the present invention, the medicine is in the form of tablets, powders, granules, capsules, pills, aqueous solutions, alcoholic solutions or syrups.

[0028] The present invention provides a dendrobium polysaccharide wine for preventing and / or treating alcohol-induced neuritis, comprising a base wine with an alcohol concentration of 30-50% and dendrobium polysaccharide; the mass volume ratio of the dendrobium polysaccharide to the base wine is 1-10g:1L.

[0029] In the present invention, 1L of base wine further includes 50-200g of flavoring agent; the base wine can be conventional commercially available liquor, and the flavoring agent is honey.

[0030] The present invention also provides a method for preparing the dendrobium polysaccharide wine, which comprises dissolving the dendrobium polysaccharide in base wine, homogenizing and aging the wine to obtain the dendrobium polysaccharide wine.

[0031] In the present invention, dendrobium polysaccharide is dissolved in base wine, and after the dendrobium polysaccharide is completely dissolved, flavoring is added and stirred until completely dissolved to obtain a solution, which is homogenized 2 to 3 times by a homogenizer. The homogenized solution is placed in a sealed container and aged in a cool and dry place for 0.5 to 1.5 months. After the aging is completed, it is filtered to obtain dendrobium polysaccharide liquor. The homogenization pressure is 20 to 30 MPa, the homogenization time is 3 to 5 min / time; the aging temperature is 15 to 25 ° C, the aging relative humidity is 60% to 75%, the aging time is preferably 1 month, and the filtration method is double filtration using gauze and filter paper, the gauze is 4 layers of gauze, the filter paper is preferably coffee filter paper, and the pore size of the filter paper is 20 to 30 μm.

[0032] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0033] Example Preparation of Dendrobium Polysaccharide Wine (DOP)

[0034] 1. Raw materials Dendrobium polysaccharide (BR, 60%, Shanghai Yuanye Biotechnology Co., Ltd., product number: S27808), liquor (52%vol Fupingchun liquor, Henan Fupingchun Liquor Co., Ltd.), honey.

[0035] 2. Preparation process Purified water is used to adjust the alcohol content of liquor to 38° (i.e., the alcohol concentration is 38% vol) as the base liquor; 1 gram of dendrobium polysaccharide powder and 50 grams of honey are added to 1 liter of the base liquor, and the dendrobium polysaccharide powder is first slowly added to the base liquor while stirring until it is fully dissolved, and then honey is added as a flavoring to improve the flavor and taste of the liquor, and stirred until it is completely dissolved to obtain a solution; a homogenizer is used to homogenize the solution at a pressure of 20 MPa for 5 minutes and 3 times to make the solution more delicate and uniform; the homogenized solution is placed in a sealed container and aged in a cool place (temperature: 15-25°C, relative humidity: 60%-75%) for 1 month to fully integrate the ingredients and make the taste more mellow. After aging, double filter with 4 layers of gauze + coffee filter paper (purchased from pakchoice flagship store, PAKCHOICE hand-brew coffee filter paper, 100 pieces of fan-shaped V-shaped filter paper, 100 pieces of conical 02 filter paper for coffee machine) to remove possible impurities and obtain the finished product of Dendrobium polysaccharide wine.

[0036] Experimental example

[0037] 1. Grouping and liquor induction model construction Six-week-old C57BL / 6J mice were purchased from Beijing Huafukang Biotechnology Co., Ltd. (Beijing, China). After one week of acclimation in the animal room, the mice were randomly divided according to body weight into a blank control group (BLK), a model group (E_ALC), a positive control group (LU; China Jinjiu, purchased from Hubei Jinpai Health Wine Co., Ltd.), and a Dendrobium Polysaccharide Liquor group (DOP), with eight mice in each group. Except for the blank control group, mice in all other groups were given ethanol by gavage (3.6 g ethanol / kg / day) for four weeks to establish a long-term alcohol exposure model (baijiu-induced model). Subsequently, mice in each group received the following intervention treatments for four weeks: The Chinese Dietary Guidelines state that male adults can consume 25g of ethanol per day, with an alcohol content of 38% alcohol. Based on the relationship between the dosage multiples for experimental animals and humans (the mouse dosage is typically 8.65 times the human dosage), the gavage dose for each mouse group was calculated: blank control group (3.6g saline / kg / day), model group (3.6g ethanol / kg / day), positive control group (3.6g ethanol / kg / day), and Dendrobium polysaccharide liquor group (3.6g Dendrobium polysaccharide liquor obtained in Example 1 / kg / day). The baijiu and Jinjiu liquor used in the above experiments were reduced to 38% alcohol with purified water and then administered daily based on the weight of each mouse group and the gavage dose standard. Neurobehavioral testing was performed after the four-week intervention, and brain tissue, hippocampus, and other experimental samples were subsequently collected from the mice for subsequent testing of neuroinflammatory markers.

[0038] 2. Neuroethology

[0039] 2.1 Y-maze Test The Y-maze test is a behavioral test method commonly used to evaluate the spatial memory and cognitive function of animals. This experiment uses frosted non-transparent organic plastic, made in the shape of a "Y", divided into three arms: A, B, and C (35 cm long, 10 cm wide, and 20 cm high), with the angles between the three arms being 120°. At the beginning of the experiment, the mouse is placed at the junction of the three arms in the same direction and allowed to shuttle through the device. The order in which the mouse enters each arm is recorded within 5 minutes. A mouse that enters two-thirds or more of each arm is counted as a successful entry into the arm, otherwise it is not counted as an entry. In order to eliminate the odor between animals and ensure the environmental hygiene of the experimental equipment, 75% alcohol is used to wipe it before and after the experiment. The next round of experiments will be carried out after the alcohol evaporates. The smell and residual alcohol will affect the behavioral results of the experimental animals. After the experiment, the order of entry into each arm is analyzed, and the correct transition is recorded in the form of "ABC", "ACB", "BCA", "BAC", "CAB", and "CBA". The spontaneous alternation rate index in the Y-maze experiment can intuitively reflect the extent of the impact of neuroinflammation on the cognitive function of mice, especially spatial memory. The results are shown in Figure 2. Figure 1 .

[0040] Depend on Figure 1 As shown in a, the spontaneous alternation rate of mice in the E_ALC group in the Y maze was significantly lower than that in the BLK group and the DOP group ( P <0.05), and there was no significant difference between the DOP group and the BLK group, indicating that DOP intervention can improve the symptoms of spatial memory decline in mice with long-term alcohol consumption.

[0041] 2.2 Open field test (OPT) The open field experiment can be used to evaluate the changes in cognitive function and anxiety in mice after long-term exposure to alcohol. The mice were placed individually in a 50cm*50cm*50cm opaque square medical ABS material experimental box, and tracking software was used to record and analyze the spontaneous activities of the mice, such as movement trajectory, movement distance, movement time, stagnation time, etc. The activities of the animals in the box were automatically recorded. The experimental time was usually 5 minutes. In order to avoid interference from previous experiments, the experimental device was cleaned with 75% ethanol after each experiment. The central area residence time index of the open field test reflects the changes in the anxiety-like behavior and exploratory behavior of mice. The results are shown in Figure 1 .

[0042] Depend on Figure 1 As shown in b, the time that mice in the E_ALC group stayed in the central area was significantly lower than that in the BLK group ( P <0.05), after DOP intervention, the time mice spent in the central area increased significantly ( P <0.05) and returned to normal levels, indicating that DOP can improve the anxiety and exploratory behavior of mice treated with liquor.

[0043] 2.3 Rotarod test The fatigue rotarod test can be used to assess the motor coordination and balance abilities of mice chronically exposed to alcohol. In the experiment, a group of mice is placed on a channel (if it is difficult to handle the mice, leave an empty channel between two mice and only measure 3 mice per run). Try to get the mice on the rod to walk forward to maintain balance. The rod initially rotates at a constant speed of 4 rpm to allow all mice to be positioned in their respective channels. Once all the mice are "ready" (i.e., check whether they can walk forward at 4 rpm for a few seconds), press the start button and the rod will accelerate from 4 rpm to 40 rpm within 300 seconds. During the entire experiment, the rotarod instrument automatically records the latency, speed and movement distance of each mouse falling from the rod, and records the reason for the end of the test (falling, jumping, passive rotation). After a group of experiments, the equipment is cleaned with water and then wiped dry with 75% ethanol, and then the above steps are repeated to test the remaining mice. Neuroinflammatory response interferes with the transmission of nerve signals, affects the contraction and relaxation function of muscles, and thus leads to a decrease in the animal's exercise endurance, which is manifested as a shortened movement distance in the fatigue rotarod test. The results are shown in the figure. Figure 1 .

[0044] like Figure 1 As shown in c, the movement distance of mice in the E_ALC group was significantly lower than that of the BLK group ( P <0.05), after DOP intervention, the movement distance of mice increased significantly ( P <0.05) and returned to normal levels, indicating that DOP can restore the exercise capacity of mice with long-term alcohol consumption.

[0045] In summary, Dendrobium polysaccharide wine can improve neurological dysfunctions such as cognitive dysfunction, decreased motor coordination ability, and anxiety-like behavior in animals caused by long-term drinking, suggesting that it has a comprehensive protective effect against alcoholic nerve damage.

[0046] 3. Physiological and biochemical index detection

[0047] 3.1 Determination of inflammatory factors in brain tissue Inflammatory factors can affect the blood-brain barrier and neurodegeneration. In this study, ELISA kits (Merck Biotech Co., Ltd.) were used to detect the levels of neuroinflammatory factors such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in mouse brain tissue. The experimental procedures were strictly carried out according to the instructions provided by the ELISA kit manufacturer. The results are shown in Figure 2. Figure 2 .

[0048] like Figure 2As shown in the results, the levels of inflammatory factors such as IL-6, IL-1β and TNF-α in the brain of mice in the E_ALC group exposed to white wine for a long time were significantly higher than those in the BLK group ( P <0.05), after DOP intervention, the levels of IL-1β and TNF-α in the mouse brain decreased significantly ( P <0.05), and returned to the level of the BLK group, indicating that DOP inhibited the expression of inflammatory factors in the brains of long-term alcohol-drinking mice.

[0049] 3.2. Determination of neurotransmitters in the hippocampus The present invention uses ultra-performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UPLC-Q-TOF-MS) to determine the content of neurotransmitters such as 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), glutamate (Glu), and dopamine (DA) in the hippocampus. Figure 3 .

[0050] Mouse hippocampus sample preparation method: Mice were decapitated and the brains were immediately removed on ice. Rinse in ice-cold saline, drain the saline with filter paper, and the brains were placed on ice to quickly dissect the hippocampus and weigh it. Ice-cold 0.1% formic acid was added to the hippocampus and homogenized at a ratio of 1:10 (mg / mL). The mixture was centrifuged at 12,000 rpm for 20 minutes at 4°C. The supernatant was collected and filtered through a 0.22 μm microporous filter membrane for analysis.

[0051] Preparation of standard solutions: Weigh approximately 5.0 mg of 5-HT, GABA, Glu, and DA standards, dissolve each in solvent (0.2% formic acid-methanol, 8:2, V / V), and dilute to 5 mL to prepare standard stock solutions. Mixed standards are then prepared.

[0052] Liquid chromatography-mass spectrometry conditions: ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 μm); Mobile phase A: 0.1% formic acid in water; Mobile phase B: acetonitrile; Gradient elution program: 0-2.5 min, 5% B, 2.5-4 min, 5%-20% B; 4-7 min, 20%-60% B; 7-7.5 min, 60%-5% B; 7.5-10 min, 5% B. Flow rate: 0.2 mL / min. Column temperature: 30°C. Injection volume: 3 μL.

[0053] MS conditions: Mass spectrometry conditions electrospray ionization source (ESI): positive ion mode; multiple reaction monitoring mode (MRM) curtain gas: 40.0 psi; collision gas: N2; spray voltage: 5500 V; ion source temperature: 500°C; GS1: 50 psi, GS2: 50 psi; gas flow rate: 12 L / min.

[0054] Depend on Figure 3 It can be seen that compared with the blank control group, long-term exposure to alcohol treatment significantly reduced ( P <0.05) The levels of 5-HT, DA, and GABA in the hippocampus were significantly increased ( P <0.05) in the hippocampus. After DOP intervention, the alcohol-induced decrease in DA content in the hippocampus was significantly reversed ( P <0.05) and the increase of Glu content ( P <0.05), although the contents of 5-HT and GABA did not decrease significantly, there was no statistical difference compared with the BLK group. The results showed that DOP can regulate the concentrations of neurotransmitters such as 5-HT, DA, GABA and Glu in the hippocampus of long-term alcohol-drinking mice.

[0055] 3.3. HE staining of mouse hippocampal tissue The brain tissue was removed along the coronal plane to preserve the intact hippocampal tissue, fixed with 10% formaldehyde solution, embedded, dehydrated, transparentized with xylene, stained with HE, and sealed with neutral resin by an automatic sealing machine. The slides were observed under an optical microscope. The results are shown in the figure. Figure 4 .

[0056] HE staining was used to examine the effect of DOP on the structure of the CA3 region of the hippocampus in alcohol-exposed mice. Figure 4 As shown, the number of neurons in the CA3 region of the hippocampus of mice in the BLK group consisted of 4-5 layers of cells. Cell morphology was normal, with intact cell bodies, prominent nucleoli, and a tightly packed, neat arrangement. The number of pyramidal cells in the CA3 region of mice in the E_ALC group was significantly reduced, and these cells exhibited various abnormalities, including swollen cell bodies, cytoplasmic vacuoles, absent or marginalized Nissl bodies, and pyknotic nuclei. After DOP treatment, the number of neurons in the CA3 region increased, and the morphology of most pyramidal cells returned to normal, with narrowed intercellular spaces and a return to a tightly packed, neat arrangement. This suggests that DOP can alleviate the inflammatory damage to neurons caused by long-term alcohol consumption and restore the damaged hippocampal structure.

[0057] 3.4 Determination of oxidative markers in the brain Brain samples were removed from a -80°C freezer and homogenized with physiological saline to a 10% mass fraction. Liver and brain supernatants were then centrifuged at 4°C and 3500 rpm for 10 min. The AST and ALT levels in the liver, as well as the reduced glutathione (GSH) content (to assess the extent of neuronal damage), malondialdehyde (MDA) content (to reflect the extent of lipid peroxidation), and superoxide dismutase (T-SOD) activity (to measure antioxidant capacity) in the liver and brain tissue homogenates were determined according to the kit instructions. The results are shown in Figure 5 .

[0058] DOP can alleviate the imbalance of the oxidative system in the brain of mice that drink alcohol for a long time. Figure 5 As shown in Figure 2, compared with the BLK group, the levels of oxidative stress indicators such as GSH and T-SOD in the brain of mice in the E_ALC group were significantly reduced ( P <0.05), MDA content increased significantly ( P <0.05), after DOP intervention, the levels of GSH and T-SOD in the mouse brain increased significantly ( P <0.05), MDA content was significantly decreased ( P <0.05) were not significantly different from those in the BLK group.

[0059] In summary, DOP can intervene in neuroinflammation in long-term alcohol-drinking mice by changing their neurobehavior, inflammatory factors, neurotransmitters, hippocampal structure, oxidative stress and other physiological and biochemical indicators.

[0060] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. Use of dendrobium polysaccharide in the preparation of a medicament for preventing and / or treating alcohol-induced neuroinflammation, characterized in that: The alcohol-induced neuroinflammation is an alcoholic neurological disease caused by excessive drinking.

2. The use according to claim 1, characterized in that The alcoholic neurological diseases caused by excessive drinking include alcoholic encephalopathy and neurodegenerative diseases caused by excessive drinking.

3. The use according to claim 1 or 2, characterized in that The medicine is in the form of tablets, powders, granules, capsules, pills, liquids, wines or syrups.

4. A dendrobium polysaccharide wine for preventing and / or treating alcohol-induced neuritis, characterized in that: The invention comprises base wine with an alcohol concentration of 30-50% and dendrobium polysaccharide; the mass volume ratio of the dendrobium polysaccharide to the base wine is 1-10g:1L.

5. The dendrobium polysaccharide wine according to claim 4, characterized in that 1L of base wine also includes 50~200g of flavoring.

6. The dendrobium polysaccharide wine according to claim 5, characterized in that The flavoring agent is honey.

7. The method for preparing the dendrobium polysaccharide wine according to any one of claims 4 to 6, characterized in that: The dendrobium polysaccharide is dissolved in the base wine, homogenized and aged to obtain the dendrobium polysaccharide wine.

8. The preparation method according to claim 7, characterized in that The homogenization pressure is 20-30 MPa, the number of homogenizations is 2-3 times, and the homogenization time is 3-5 min / time.

9. The preparation method according to claim 7, characterized in that The aging temperature is 15-25° C., the relative humidity is 60%-75%, and the aging time is 0.5-1.5 months.

10. The preparation method according to claim 7, characterized in that Before most homogenization, a flavoring agent step is also included, and after the aging is completed, a filtering operation is also included.

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