Polygonatum sibiricum polysaccharide with effects of dispelling effects of alcohol and protecting liver as well as preparation method and application of polygonatum sibiricum polysaccharide

Abstract

The invention belongs to the technical field of biological pharmacy, and particularly relates to rhizoma polygonati polysaccharide with the effects of preventing acute alcoholism, dispelling the effects of alcohol and protecting the liver as well as a preparation method and application of the rhizoma polygonati polysaccharide. The method comprises the following steps: crushing dried rhizoma polygonati in a crusher, taking a certain amount of rhizoma polygonati powder, adding distilled water according to a set material-liquid ratio, adjusting the pH value to 5.0, adding 1% of compound enzyme, putting into a water bath kettle, carrying out ultrasonic-assisted extraction at 65 DEG C for 55 minutes, carrying out enzyme deactivation at 95 DEG C for 10 minutes, filtering while hot, putting filtrate into a rotary evaporator, carrying out evaporation concentration, adding a Sevage reagent with the volume 4 times that of the filtrate, shaking for 20 minutes, centrifuging to remove protein, and dialyzing with deionized water for 24 hours, and adding 95% ethanol, standing overnight, centrifuging for 15 minutes at the speed of 3500 r / min, and freeze-drying the precipitate to obtain the polygonatum polysaccharide. Polygonatum sibiricum polysaccharide can effectively enhance the activity of ethanol dehydrogenase and acetaldehyde dehydrogenase and promote catabolism of alcohol.

Description

Technical Field

[0001] The present invention belongs to the technical field of biopharmaceuticals, and particularly relates to a polygonatum polysaccharide having the functions of preventing acute alcoholism and protecting the liver from alcohol damage, and a preparation method and application thereof. Background Art

[0002] Alcoholic liver disease (ALD) is a disease caused by liver damage due to excessive alcohol consumption over a long period. Excessive alcohol consumption can cause acute alcohol poisoning, which can cause irreversible harm to organs such as the liver, damage the body's functions, or change the central nervous system from an excited state to an inhibitory state, thereby triggering a series of diseases. The situation of adults suffering from alcoholic liver injury in China is relatively common, and the patient proportion is between 4.3% and 6.5%. When alcohol enters the human body, it will stimulate liver cells to undergo oxidative stress reactions and the accumulation of reactive oxygen species, leading to lipid peroxidation and further damaging organs. After alcohol enters the human body through the mouth, it first enters the stomach via the esophagus, then reaches the liver through the blood, is absorbed and decomposed by the liver, and finally is excreted out of the body through urine, sweat, and respiration.

[0003] The liver is the main organ for ethanol metabolism. The decomposition process of ethanol can only proceed under the catalytic action of alcohol dehydrogenase (ADH), and is first converted into acetaldehyde, and then catalyzed by aldehyde dehydrogenase (ALDH) to acetic acid. Acetic acid enters the tricarboxylic acid cycle under the action of acetyl coenzyme A and finally becomes CO2 and H2O and is excreted out of the body. Therefore, ADH and ALDH play very important roles in the decomposition of alcohol.

[0004] It is generally believed that the pathogenesis of ALD is oxidative stress. While alcohol metabolism increases the burden on the liver, a large number of free radicals are generated, leading to oxidative damage to the liver. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX) provide antioxidant protection. During the process of oxidative stress, SOD can scavenge superoxide anion free radicals harmful to the body; CAT catalyzes the decomposition of hydrogen peroxide into oxygen molecules and water, thereby providing antioxidant protection in the body; GSH-PX is also an important peroxide-decomposing enzyme, catalyzing GSH to become GSSG, reducing toxic peroxides to non-toxic hydroxy compounds, and at the same time promoting the decomposition of H2O2, thereby protecting the structure and function of cell membranes from the interference and damage of peroxides. The content of MDA can reflect the degree of lipid peroxidation damage.

[0005] The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (AKP) are important indicators reflecting the liver function status. ALT and AST respectively catalyze the transamination between glutamate and pyruvate, and between glutamate and oxaloacetate. AKP participates in the phosphorylation and dephosphorylation reactions of substrates and plays an important regulatory role in cell metabolism and signal transduction. When liver tissue undergoes lesions such as hepatitis, liver cancer, cirrhosis, etc., the activities of these enzymes will increase abnormally.

[0006] At present, most liver-protecting drugs are chemically synthesized compounds, which have certain toxic and side effects. Excessive consumption or improper use will increase the burden and damage to the liver. Therefore, it is of great value and significance to find natural and non-toxic drugs for relieving alcohol intoxication and protecting the liver.

[0007] Polygonatum sibiricum is a perennial herb of the genus Polygonatum in the family Liliaceae. Its dried rhizome is used as medicine, and it has the effects of invigorating qi and nourishing yin, strengthening the spleen, moistening the lungs, and tonifying the kidneys. Polysaccharide is the main active ingredient with a relatively high content in Polygonatum sibiricum, which has the effects of reducing blood lipid, reducing blood sugar, antioxidation and anti-fatigue, etc. However, there is no research and report on the effect of Polygonatum sibiricum polysaccharide in relieving alcohol intoxication and protecting the liver. Summary of the Invention

[0008] To solve the problems existing in the prior art, the present invention provides a Polygonatum sibiricum polysaccharide with the functions of treating acute alcohol intoxication and relieving alcohol intoxication and protecting the liver, its preparation method and application.

[0009] The technical solution adopted by the present invention to solve the technical problems is as follows:

[0010] The present invention provides a preparation method of a Polygonatum sibiricum polysaccharide with the functions of treating acute alcohol intoxication and relieving alcohol intoxication and protecting the liver, which mainly includes the following steps:

[0011] Take the dried Polygonatum sibiricum and place it in a pulverizer for pulverization. Take a certain amount of Polygonatum sibiricum powder, add distilled water according to the set solid-liquid ratio, adjust the pH to 5.0, add 1% compound enzyme (papain + cellulase), place it in a water bath, extract with ultrasonic wave at 65°C for 55 min, then inactivate the enzyme at 95°C for 10 min, filter while it is hot, place the filtrate in a rotary evaporator for evaporation and concentration, add 4 times the volume of Sevage reagent, shake for 20 min, centrifuge to remove proteins, dialyze with deionized water for 24 h, add 95% ethanol, let it stand overnight, centrifuge at 3500 r / min for 15 min, and freeze-dry the precipitate to obtain Polygonatum sibiricum polysaccharide.

[0012] Preferably, take Polygonatum sibiricum and place it in a pulverizer for pulverization and pass through a 55-65 mesh sieve.

[0013] Preferably, the solid-liquid ratio is 1:(30-35) g / mL.

[0014] Preferably, the pH of the extract is adjusted to 4.5-5.5.

[0015] Preferably, the mass ratio of papain to cellulase in the mixture is 6:4.

[0016] Preferably, the filtrate is placed in a rotary evaporator and evaporated and concentrated at 160 - 170 rpm and 45 - 55 °C.

[0017] Preferably, after removing the protein, the centrifuged liquid is placed in a dialysis bag with a molecular weight cut-off of 0.45 - 0.55 KDa for 20 - 30 h to remove salts.

[0018] Preferably, 3.5 - 4.5 times the volume of 95% ethanol is added and then left to stand overnight at 4 °C.

[0019] A polygonatum polysaccharide with the functions of treating acute alcoholism and protecting the liver from alcohol damage, prepared by the preparation method of a polygonatum polysaccharide with the functions of treating acute alcoholism and protecting the liver from alcohol damage provided by the present invention.

[0020] A polygonatum polysaccharide with the functions of treating acute alcoholism and protecting the liver from alcohol damage, prepared by the preparation method of a polygonatum polysaccharide with the functions of treating acute alcoholism and protecting the liver from alcohol damage provided by the present invention, is applied to the preparation of drugs for treating acute alcoholism and drugs for protecting the liver from alcohol damage.

[0021] The beneficial effects of the present invention are as follows:

[0022] The polygonatum polysaccharide prepared by the present invention exerts its functions and effects in the following aspects:

[0023] 1. The polygonatum polysaccharide can effectively enhance the activities of ADH and ALDH, promote the catabolism of alcohol, and thus reduce the alcohol content in the liver and blood of rats.

[0024] 2. The polygonatum polysaccharide has strong antioxidant activity, improves the activities of SOD, CAT and GSH-PX in the liver and blood of rats, enhances the antioxidant and free radical scavenging abilities of the rat liver, and reduces the content of lipid peroxidation product MDA in the rat liver.

[0025] 3. The polygonatum polysaccharide can maintain the normal activities of AST, ALT and AKP in the liver of rats, and effectively prevent the liver biochemical function damage caused by acute alcoholism.

[0026] 4. The polygonatum polysaccharide can effectively maintain the liver tissue structure of rats and effectively prevent the liver tissue structure lesions caused by acute alcoholism. Description of the Drawings

[0027] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the accompanying drawings required in the embodiments. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other accompanying drawings can be obtained based on these drawings.

[0028] Figure 1 Determination of the relative molecular mass of polygonatum polysaccharide by gel permeation high performance liquid chromatography;

[0029] Figure 2 Chromatograms of the monosaccharide standard mixture solution (a) and polygonatum polysaccharide (b);

[0030] Figure 3 Infrared spectrum of polygonatum kingianum polysaccharide;

[0031] Figure 4 Staining of rat liver sections (200); among them, (A) blank group, (B) model group, (C) bifendate group, (D) high-dose polygonatum polysaccharide group, (E) low-dose polygonatum polysaccharide group;

[0032] Figure 5 Effect of polygonatum polysaccharide on the activities of AST, ALT, and AKP in rat liver;

[0033] Figure 6 Activities of ADH and ALDH in rat liver and serum ethanol concentration;

[0034] Figure 7 Activities of SOD, CAT, and GSH-PX and MDA content in rat liver;

[0035] Figure 8 Effect of polygonatum polysaccharide on the activities of SOD, CAT, and GSH-PX and MDA content in rat serum. Detailed implementation manners

[0036] Now, various exemplary implementation manners of the present invention will be described in detail. Unless otherwise specified, the methods in the embodiments are all conventional methods, and the reagents used are all conventional commercially available reagents or reagents prepared by conventional methods. This detailed description should not be regarded as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics, and implementation schemes of the present invention.

[0037] It should be understood that the terms described in the present invention are only for describing specific embodiments and are not intended to limit the present invention. Additionally, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0038] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. Although the present invention only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention.

[0039] Without departing from the scope or spirit of the present invention, various modifications and variations can be made to the specific embodiments of the present invention specification, which will be obvious to those skilled in the art. Other embodiments obtained from the present invention specification will be obvious to those skilled in the art. The present invention specification and examples are merely exemplary.

[0040] Regarding the use of "comprising", "including", "having", "containing", etc. herein, they are all open-ended terms, meaning including but not limited to.

[0041] 1. Extraction, Preparation and Characterization Analysis of Polygonatum sibiricum Polysaccharide

[0042] 1.1. Extraction and Preparation of Polygonatum sibiricum Polysaccharide

[0043] The Polygonatum sibiricum was from the farmers' market in Pu'er City, Yunnan Province. It was the dried rhizome of Polygonatum kingianum Coll. et Hemsl., sliced, naturally dried in the sun, pulverized and passed through a 60-mesh sieve, then sealed and stored for later use. Take the Polygonatum sibiricum powder, add distilled water according to the solid-liquid ratio of 1:30 (g / mL), adjust the pH to 5.0, add 1% compound enzyme (papain: cellulase = 6:4), place it in a water bath, extract at 65 °C for 55 min, heat at 95 °C for 10 min to inactivate the enzyme, filter while it is hot, place the filtrate in a rotary evaporator, concentrate at 170 rpm and 50 °C, add 4 times the volume of Sevage reagent, shake for 20 min, centrifuge to remove proteins, repeat 8 times, dialyze with deionized water for 24 h, add 4 times the volume of ethanol, let it stand overnight at 4 °C, centrifuge at 3500 r / min for 15 min, and freeze-dry the precipitate to obtain the Polygonatum sibiricum polysaccharide extract.

[0044] 1.2. Determination of the Content of Polygonatum sibiricum Polysaccharide by Phenol-Sulfate Method

[0045] Accurately measure 0.00 mL, 0.20 mL, 0.40 mL, 0.60 mL, 0.80 mL, and 1.00 mL of 1.00 mg / mL glucose standard solution, add distilled water to make up to 3.00 mL, add 2.00 mL of 5% phenol and 5.00 mL of concentrated sulfuric acid, shake well, heat in a boiling water bath for 15 min, take out, cool to room temperature with tap water, measure the absorbance of the glucose standard solution with a microplate reader at a wavelength of 490 nm, make a standard curve with the absorbance value as the ordinate and the glucose concentration as the abscissa, and calculate the regression equation and the correlation coefficient (R 2 ).

[0046] Dissolve the polygonatum polysaccharide powder in water in a test tube, measure the absorbance of the sample with a microplate reader at a wavelength of 490 nm, substitute the data of three parallel samples into the linear regression equation obtained above, and calculate the content of polygonatum polysaccharide.

[0047] 1.3. Molecular weight of polygonatum polysaccharide

[0048] Use a Shimadzu LC-20AT high-performance gel permeation chromatograph and a TSKgel GMPWXL aqueous gel chromatographic column. The 5 mg / mL polygonatum polysaccharide solution is centrifuged at 12000 r / min for 10 min, filtered through a 0.22 μm microporous filter membrane, and eluted at a flow rate of 0.6 mL / min at 35 °C. Using pullulan polysaccharides with different relative molecular weights (Mw 642000, 334000, 49400, 22000, 6300) as standard samples, plot a standard curve with lg Mw against the retention time t R and calculate the molecular weight of the sample.

[0049] Plotting the lg Mw of pullulan sugar against the retention time t R gives the calibration curve equation: Y = -0.71172X + 15.08081, R 2 = 0.99299. The polysaccharide after impurity removal and purification shows a main absorption peak at 16.016 min by high performance liquid permeation chromatography. According to the retention time, the molecular weight of polygonatum polysaccharide is calculated to be 10.684 kDa( Figure 1 ).

[0050] 1.4. Monosaccharide composition of polygonatum polysaccharide

[0051] Weigh an appropriate amount of polysaccharide sample, add 1 mL of 2 mol / L trifluoroacetic acid solution, heat at 121 °C for 2 h, then dry with nitrogen, wash with methanol 2 - 3 times, dry again, and add sterile water until dissolved for measurement.

[0052] Using glucose (Glc), fucose (Fuc), rhamnose (Rha), arabinose (Ara), galactose (Gal), xylose (Xyl), mannose (Man), fructose (Fru), ribose (Rib), galacturonic acid (Gal-UA), glucuronic acid (Glc-UA), mannuronic acid (Man-UA), and guluronic acid (Gul-UA) as standard monosaccharide samples, a Thermo ICS5000 ion chromatograph and Dionex TM CarboPac TM PA20 (150*3.0mm, 10μm) liquid chromatography column. The injection volume was 5 μL, the mobile phase A was H2O, the mobile phase B was 0.1M NaOH, the mobile phase C was 0.1M NaOH and 0.2M NaAc, and the flow rate was 0.5 mL / min. The column temperature was 30 °C, and the elution gradient was as follows: at 0 min, the ratio of mobile phase A / mobile phase B / mobile phase C was (95:5:0, V / V); at 26 min, it was (85:5:10, V / V); at 42 min, it was (85:5:10, V / V); at 42.1 min, it was (60:0:40, V / V); at 52 min, it was (60:40:0, V / V); at 52.1 min, it was (95:5:0, V / V); at 60 min, it was (95:5:0, V / V).

[0053] Through chromatographic analysis, the ion chromatogram of the mixed standard sample of monosaccharides ( Figure 2 a) and the ion chromatogram of polygonatum polysaccharide ( Figure 2 b) were obtained. The results showed that polygonatum polysaccharide was composed of monosaccharides such as Glc (63.36%), Gal (16.90%), Gal-UA (11.52%), Ara (3.53%), Rib (2.14%), Man (1.35%), and Rha (1.20%), etc., with Glc, Gal, and Gal-UA being the main components.

[0054] 1.5. Chemical bond structure of polygonatum polysaccharide

[0055] Weighed 200 mg of potassium bromide dried at 105 °C to constant weight, added 2 mg of polygonatum polysaccharide, ground it evenly under infrared light in an agate mortar, and pressed it into a thin slice with a tablet press. Infrared spectrum scanning was carried out in the range of 4000 - 500 cm -1 to verify the chemical structure of the polysaccharide according to each absorption peak.

[0056] Figure 3 It can be seen that 3406 cm –1 is the absorption peak of O-H stretching vibration. 2937 cm -1 and 1336 cm -1Two obvious characteristic peaks appear here, representing the stretching vibration and deformation vibration of C-H in carbohydrates, 1622 cm –1 is the absorption peak of C=O stretching vibration. 1134 cm –1 and the absorption peaks at 1028 cm –1 are caused by the stretching vibration of the C-O-C glycosidic bond, indicating the presence of furanoid glycosidic bonds in polygonatum polysaccharide. The absorption peak at 933 cm –1 indicates the presence of pyranoid glycosidic bonds in polygonatum polysaccharide. Therefore, both furanoid and pyranoid glycosidic bonds exist simultaneously in the polygonatum polysaccharide molecule.

[0057] 2. Protective effect of polygonatum polysaccharide on acute alcoholic liver injury in rats

[0058] 2.1. Rat test model

[0059] SPF-grade rats, male, weighing 200±20 g, feeding conditions: temperature 20-25°C, humidity 45% - 55%, 12 h of light and 12 h of darkness each. Animal experiment license number: SCXK(Dian)K2020-0004. 40 male SD rats were adaptively fed for 14 d and randomly divided into 5 groups, namely the blank group, the model group, the bifendate group, the high-dose polygonatum polysaccharide group (HPP), and the low-dose polygonatum polysaccharide group (LPP), with 8 rats in each group. The blank group and the model group were intragastrically administered normal saline, and the bifendate group (4 mg / kg body weight), the low-dose group (245 mg / kg body weight), and the high-dose group (490 mg / kg body weight) were intragastrically administered the corresponding drugs or polygonatum polysaccharide; 1 h after administration, except for the blank group, all were intragastrically administered 50% ethanol at 1 mL / 100 g (body weight), and the rest were normally fed and given water. Intragastric administration of the drug was continued for 14 d.

[0060] 2.2. Prevention of acute alcoholic liver lesions in rats by polygonatum polysaccharide

[0061] After anesthetizing and sacrificing the rats, their livers were taken, fixed with 10% formaldehyde, dehydrated with gradient ethanol, cleared with xylene for 30 min, embedded in paraffin for sectioning, and baked at 60°C. The sections were passed through the xylene solution 3 times, 10 min each time, dewaxed with gradient ethanol for 5 min, washed with distilled water, stained with hematoxylin, washed with water, differentiated with 1% hydrochloric acid solution for several seconds, blued with 0.6% ammonia water after washing, immersed in 0.5% eosin staining solution for 1-3 min, dehydrated and cleared with xylene, and sealed with neutral gum and then examined under a microscope.

[0062] Figure 4It can be seen that the cytoplasm of liver tissue cells in the blank group of rats was rich, the hepatic sinusoids were not dilated, and no obvious inflammatory cells were found in the liver parenchyma (A). In the liver tissue of the model group of rats, a small amount of fibrous tissue hyperplasia, mild edema, dilation of hepatic sinusoids were visible, and inflammatory cells appeared, showing obvious characteristics of alcoholism (B). In the bifendate group of rats, some hepatocytes showed mild edema, but no obvious inflammatory cells were found (C). In the high-dose polygonatum polysaccharide group of rats, some hepatocytes showed mild edema, and the hepatic sinusoids were not dilated (D). In the low-dose polygonatum polysaccharide group of rats, some hepatocytes showed mild edema, and a small amount of inflammatory cells were visible (E). The liver tissue structure of rats showed that bifendate had a significant effect on the prevention and treatment of acute alcoholism, and polygonatum polysaccharide also showed good effects.

[0063] 2.3. Protective effect of polygonatum polysaccharide on the biochemical function of rat liver

[0064] After the rats were anesthetized and sacrificed, liver tissues were taken and homogenized for sampling. According to the determination method described in the kit instructions, the activities of AST, ALT and AKP in the rat liver were detected.

[0065] Figure 5 It can be seen that the activities of AST, ALT and AKP in the liver of the model group of rats with acute alcoholism were abnormally increased, significantly higher than those of the blank group. Compared with the rats in the model group, the activities of ALT, AST and AKP in the livers of the rats administered bifendate and polygonatum polysaccharide decreased significantly and basically maintained at the level of the rats in the blank group. The test results showed that the liver function of the rats with acute alcoholism was severely damaged, and pretreatment with polygonatum polysaccharide could effectively prevent this damage. The test results of the biochemical indexes of liver function also verified the microscopic observation results of liver tissue sections again, and also showed that the organic lesions of the liver with acute alcoholism were basically consistent with the abnormal increase in the activities of AST, ALT and AKP in the liver.

[0066] 2.4. Polygonatum polysaccharide promotes ethanol metabolism

[0067] After the rats were anesthetized and sacrificed, blood was collected from the abdominal aorta, centrifuged at 3000 r / min to obtain serum, and liver tissue was homogenized for sampling. According to the determination method described in the kit instructions, the activities of ADH and ALDH in the rat liver were detected, and the serum ethanol concentration was measured.

[0068] Figure 6 It can be seen that compared with the blank group, the activities of ADH and ALDH in the livers of the rats in the model group were significantly increased, indicating that alcohol can activate ADH and ALDH. However, polygonatum polysaccharide can enhance the activities of ADH and ALDH, accelerate the decomposition of ethanol in the rats, and thus significantly reduce the serum ethanol concentration.

[0069] 2.5. Antioxidant effect of polygonatum polysaccharide

[0070] 2.5.1. Effects of Polygonatum sibiricum Polysaccharide on SOD, CAT, and GSH-PX in Rat Liver

[0071] After the rats were anesthetized and sacrificed, liver tissues were removed and homogenized for sampling. According to the determination method described in the kit instructions, the activities of antioxidant enzymes such as SOD, CAT, and GSH-PX in the rat liver were detected.

[0072] The effects of Polygonatum sibiricum polysaccharide on SOD, CAT, and GSH-PX in rat liver are as Figure 7 shown. Compared with the blank group, the activity levels of SOD, CAT, and GSH-PX in the liver of rats in the model group were significantly decreased; compared with the model group, administration of bifendate and high-dose Polygonatum sibiricum polysaccharide not only significantly increased the activities of SOD, CAT, and GSH-PX, but also significantly decreased the MDA content. These results indicate that similar to the effect of bifendate, Polygonatum sibiricum polysaccharide can significantly increase the activities of antioxidant enzymes such as SOD, CAT, and GSH-PX, resist oxidative stress reactions, and thus effectively prevent lipid peroxidation caused by acute alcohol intoxication.

[0073] 2.5.2. Effects of Polygonatum sibiricum Polysaccharide on SOD, CAT, and GSH-PX in Rat Serum

[0074] After the rats were anesthetized and sacrificed, blood was collected from the abdominal aorta, and serum was obtained by centrifugation at 3000 r / min. According to the determination method described in the kit instructions, the activities of antioxidant enzymes such as SOD, CAT, and GSH-PX in the rat serum were detected.

[0075] The effects of Polygonatum sibiricum polysaccharide on SOD, CAT, GSH-PX, and MDA in rat serum are as Figure 8 shown. The activities of SOD, CAT, and GSH-PX in the serum of rats in the model group were significantly lower than those in the blank group. Compared with the model group, administration of bifendate and Polygonatum sibiricum polysaccharide both significantly increased the activity levels of serum SOD, CAT, and GSH-PX, and also significantly decreased the serum MDA content. The detection results of serum antioxidant enzymes once again indicate that Polygonatum sibiricum polysaccharide can significantly increase the activity of the serum antioxidant enzyme system and effectively control the lipid peroxidation stress injury caused by acute alcohol intoxication.

[0076] The embodiments described above are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the design spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A preparation method of polygonatum polysaccharide, characterized in that, Take the dried polygonatum sibiricum and place it in a pulverizer for pulverization to obtain polygonatum sibiricum powder; take the polygonatum sibiricum powder, add distilled water according to the set solid-liquid ratio, adjust the pH to 5.0, add a composite enzyme, place it in a water bath, extract with ultrasonic assistance at 65 °C for 55 min, then inactivate the enzyme at 95 °C for 10 min, filter while it is hot, place the filtrate in a rotary evaporator for evaporation and concentration, add 4 times the volume of Sevage reagent, shake for 20 min, centrifuge to remove proteins, dialyze with deionized water for 24 h, add 95% ethanol, let stand overnight, centrifuge at 3500 r / min for 15 min, and freeze-dry the precipitate to obtain polygonatum sibiricum polysaccharide.

2. The preparation method according to claim 1, wherein The solid-liquid ratio is 1:(30 - 35) g / mL.

3. The preparation method according to claim 1, characterized in that, The composite enzyme includes papain and cellulase; the mixing ratio of papain and cellulase is 6:4 by mass ratio.

4. A polygonatum polysaccharide, characterized in that, Prepared by the preparation method according to any one of claims 1 - 3.

5. Use of the polygonatum sibiricum polysaccharide according to claim 4 in the preparation of an anti-hangover and liver-protecting drug.

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