Application of plant polysaccharide in preparation of product for dispelling effects of alcohol and protecting liver

Abstract

The invention discloses an application of plant polysaccharide in preparation of a product for dispelling effects of alcohol and protecting liver. The jasminum grandiflorum polysaccharide can be effectively used for dispelling the effects of alcohol and protecting the liver for the first time, the activity of key enzymes ADH and ALDH in an ethanol metabolism path is enhanced, the in-vivo ethanol metabolism speed is increased, liver damage caused by drunkenness is relieved, the jasminum grandiflorum polysaccharide has the body protection effect, meanwhile, the drunkenness rate can be reduced, the drunkenness time can be shortened, the content of ALT, AST, TC, TG and LDL-C in the body can be remarkably reduced, and the effect of preventing and treating the drunkenness is achieved. According to the invention, the content of HDL-C is increased, the activity of CAT, SOD and GSH in the liver of the organism is remarkably improved, the MDA level of the liver is reduced, the fat accumulation condition of the liver of the organism is remarkably improved, and a new direction is provided for preparation of products for dispelling the effects of alcohol and protecting the liver.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and particularly relates to application of frangipani polysaccharide in preparing products for sobering up and protecting the liver. Background Art

[0002] Alcoholic liver disease (ALD), caused by long-term alcohol poisoning or acute alcohol consumption, has become one of the most common liver diseases in the world. Acute alcoholic liver injury (AALI) is an early response to ALD, which usually refers to damage to the liver caused by excessive drinking in a short period of time. After alcohol enters the human body, 90% is metabolized by the liver, mainly metabolized into carbon dioxide and water by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) for excretion. However, if a large amount of alcohol is consumed in a short period of time, various disorders will occur in the body, including hepatocyte apoptosis, gastric mucosal lesions, immune cell damage, enhanced oxidative stress and lipid peroxidation.

[0003] Modern treatments for alcohol-induced acute liver injury mainly involve interventions such as corticosteroids, S-adenosylmethionine, metadoxine, silymarin, antioxidants, and restoration of intestinal function. However, silymarin, as a commonly used clinical drug for the treatment of liver injury, has significant side effects, such as nausea, vomiting, abdominal pain, loss of appetite, and diarrhea. Therefore, it is of great significance to explore bioactive substances with liver-protective effects in natural resources to reduce liver injury caused by alcoholic drinking. Summary of the invention

[0004] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention proposes an application of frangipani polysaccharide in the preparation of a product for sobering up.

[0005] The present invention also proposes the use of frangipani polysaccharide in preparing a product for preventing and / or treating liver damage caused by drinking.

[0006] The invention also provides a product.

[0007] According to a first aspect of the present invention, a use of frangipani polysaccharide in preparing a hangover sobering product is proposed.

[0008] In some embodiments of the present invention, the method for preparing the frangipani polysaccharide comprises the following steps:

[0009] The Jasmine flower sample was subjected to defatting, water extraction, alcohol precipitation, protein removal and dialysis in sequence to obtain Jasmine flower polysaccharide.

[0010] In some embodiments of the present invention, the defatting is performed using anhydrous ethanol.

[0011] In some embodiments of the present invention, in the defatting step, the solid-to-liquid ratio of the jasmine flower sample to anhydrous ethanol is 1 g: 8-12 mL.

[0012] In some embodiments of the present invention, the solid-liquid ratio is 1 g:10 mL.

[0013] In some embodiments of the present invention, degreasing is performed by heating under reflux.

[0014] In some embodiments of the present invention, the heating reflux is carried out at a temperature of 70 to 90° C. for a time of 1 to 2 hours, and the heating reflux step is repeated 2 to 3 times.

[0015] In some embodiments of the present invention, in the water extraction step, the solid-liquid ratio of the defatted jasmine flower sample to water is 1 g: 10-20 mL.

[0016] In some embodiments of the present invention, the solid-liquid ratio is 1 g:15 mL.

[0017] In some embodiments of the present invention, water extraction is performed by combining ultrasonic extraction and water bath extraction.

[0018] In some embodiments of the present invention, the ultrasonic extraction is performed under the condition of ultrasonic power of 90-110w for 15-25min.

[0019] In some embodiments of the present invention, the water bath extraction is performed at 95-105° C. for 2-3 h.

[0020] In some embodiments of the present invention, the water extraction further includes a step of solid-liquid separation and collecting the separated liquid phase.

[0021] In some embodiments of the present invention, the alcohol precipitation uses ethanol with a mass fraction of 95% or more.

[0022] In some embodiments of the present invention, in the alcohol precipitation step, the volume ratio of the water extract to ethanol is 1:(3-5).

[0023] In some embodiments of the present invention, the alcohol precipitation temperature is 0-4°C, and the alcohol precipitation time is 8-12 hours.

[0024] In some embodiments of the present invention, the step of washing the precipitate obtained by alcohol precipitation with anhydrous ethanol, ether and acetone in sequence is also included.

[0025] In some embodiments of the present invention, the protein removal is performed using Sevag reagent.

[0026] In some embodiments of the present invention, the Sevag reagent is a mixed solution of chloroform and n-butanol in a volume ratio of 4:1.

[0027] In some embodiments of the present invention, the volume ratio of the Sevag reagent to the polysaccharide solution obtained by dissolving the precipitate obtained by washing with anhydrous ethanol, ether and acetone in water is 1:4-6.

[0028] In some embodiments of the present invention, the dialysis is performed using a dialysis bag with a molecular weight cut-off of >8000Da.

[0029] In some embodiments of the present invention, the dialysis time is 1 to 3 days.

[0030] In some embodiments of the present invention, the monosaccharides in the frangipani polysaccharide consist of rhamnose, arabinose, galactose, glucose, xylose, mannose and galacturonic acid.

[0031] In some embodiments of the present invention, the molar ratio of rhamnose, arabinose, galactose, glucose, xylose, mannose and galacturonic acid is (14-22): (100-120): (160-185): (200-240): (16-25): (18-27): (400-450).

[0032] In some embodiments of the present invention, the molar ratio of rhamnose, arabinose, galactose, glucose, xylose, mannose and galacturonic acid is (17-21): (105-115): (170-180): (210-230): (18-23): (19-25): (430-440).

[0033] In some embodiments of the present invention, the molar ratio of rhamnose, arabinose, galactose, glucose, xylose, mannose and galacturonic acid is 19:109:174:220:21:22:434.

[0034] In some embodiments of the present invention, the molecular weight of the frangipani polysaccharide is 69445 Da.

[0035] According to a second aspect of the present invention, a use of Jasmine flower polysaccharide in preparing a product for preventing and / or treating liver damage caused by drinking is proposed.

[0036] In some embodiments of the present invention, the alcohol-induced liver damage includes alcohol-induced acute liver damage.

[0037] In some embodiments of the present invention, the product has at least one of the following functions:

[0038] (1) Accelerate the metabolism of ethanol and / or acetaldehyde;

[0039] (2) shorten the duration of intoxication and / or sobriety;

[0040] (3) Reduce the rate of drunkenness;

[0041] (4) Reduce liver index;

[0042] (5) Increase the activity of alcohol dehydrogenase (ADH) and / or acetaldehyde dehydrogenase (ALDH) in the body;

[0043] (6) Reduction of serum alanine aminotransferase (ALT) and / or aspartate aminotransferase (AST) levels;

[0044] (7) Increase the level of high-density lipoprotein cholesterol (HDL-C) in serum;

[0045] (8) increasing the activity of catalase (CAT), superoxide dismutase (SOD) and / or glutathione peroxidase (GSH);

[0046] (9) Reduce malondialdehyde (MDA) content;

[0047] (10) Improve ethanol-induced abnormal lipid accumulation;

[0048] (11) Prevent alcohol metabolism disorders caused by excessive alcohol intake.

[0049] In some embodiments of the present invention, increasing the ADH and ALDH enzyme activities in vivo includes increasing the ADH and ALDH enzyme activities in the liver.

[0050] In some embodiments of the present invention, the improvement of ethanol-induced abnormal lipid accumulation includes reducing the levels of triglycerides (TG), cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) in serum.

[0051] According to a third aspect of the present invention, a product for sobering up or preventing liver damage caused by drinking is provided, wherein the product comprises the above-mentioned frangipani polysaccharide.

[0052] In some embodiments of the present invention, the product is at least one of a medicine, a health product and a functional food.

[0053] In some embodiments of the present invention, the product is a medicine, and the medicine further includes a pharmaceutically acceptable excipient.

[0054] In some embodiments of the present invention, the pharmaceutically acceptable excipients include at least one of a diluent, an excipient, a filler, a binder, a disintegrant, an absorption enhancer, a surfactant, an adsorption carrier, a lubricant, a sweetener and a flavoring agent.

[0055] In some embodiments of the invention, the excipient comprises water.

[0056] In some embodiments of the present invention, the filler comprises at least one of starch and sucrose.

[0057] In some embodiments of the present invention, the binder includes at least one of a cellulose derivative, alginate, gelatin and polyvinyl pyrrolidone.

[0058] In some embodiments of the invention, the humectant comprises glycerin.

[0059] In some embodiments of the present invention, the disintegrant comprises at least one of agar, calcium carbonate and sodium bicarbonate.

[0060] In some embodiments of the invention, the absorption enhancer comprises a quaternary ammonium compound.

[0061] In some embodiments of the invention, the surfactant comprises cetyl alcohol.

[0062] In some embodiments of the present invention, the adsorption carrier includes at least one of kaolin and bentonite.

[0063] In some embodiments of the present invention, the lubricant includes at least one of talc, calcium stearate, magnesium stearate and polyethylene glycol.

[0064] In some embodiments of the present invention, the dosage form of the drug is in the form of solid, semi-solid or liquid, and can be an aqueous solution, non-aqueous solution or suspension.

[0065] In some embodiments of the present invention, the dosage form of the drug is tablets, capsules, soft capsules, granules, pills, oral liquids, dry suspensions, pellets, dry extracts, injections or infusions, transdermal agents, and transdermal microneedles.

[0066] In some embodiments of the present invention, the drug may be administered by any conventional method in the art, including but not limited to injection or oral administration.

[0067] In some embodiments of the present invention, the injection administration can be intravenous injection, intramuscular injection, intraperitoneal injection, intradermal injection or subcutaneous injection.

[0068] In some embodiments of the present invention, the mass fraction of the frangipani polysaccharide in the product is 0.01% to 100%.

[0069] According to some preferred embodiments of the present invention, the mass fraction of the frangipani polysaccharide in the product is 0.05% to 95%.

[0070] According to some preferred embodiments of the present invention, the mass fraction of the frangipani polysaccharide in the product is 0.05% to 50%.

[0071] According to some embodiments of the present invention, at least the following beneficial effects are achieved: the present invention proposes for the first time that jasmine flower polysaccharides can be effectively used for hangover and liver protection, including enhancing the activity of key enzymes ADH and ALDH in the ethanol metabolic pathway, accelerating the rate of ethanol metabolism in the body, and reducing liver damage caused by drunkenness, thereby protecting the body. At the same time, it can reduce the drunkenness rate, shorten the drunkenness time, significantly reduce the body's ALT, AST, TC, TG and LDL-C levels, increase the HDL-C content, significantly increase the body's liver CAT, SOD, GSH activities, reduce the liver's MDA content, and significantly improve the body's liver fat accumulation, providing a new direction for the preparation of hangover and liver protection products. BRIEF DESCRIPTION OF THE DRAWINGS

[0072] The present invention will be further described below with reference to the accompanying drawings and embodiments, wherein:

[0073] Figure 1 The glucose standard curve diagram in Example 1 of the present invention;

[0074] Figure 2 It is the ultraviolet spectrum analysis diagram in Example 1 of the present invention;

[0075] Figure 3 This is an infrared spectrum analysis diagram in Example 1 of the present invention;

[0076] Figure 4 This is a high performance gel permeation chromatography analysis chart in Example 1 of the present invention;

[0077] Figure 5 This is a graph showing the monosaccharide composition of the frangipani polysaccharide in Example 1 of the present invention;

[0078] Figure 6 The figure is a graph showing the test results of the weight and drunkenness rate of mice in Example 2 of the present invention, wherein A is a graph showing the test results of the weight of mice, and B is a graph showing the test results of the drunkenness rate, ###p<0.001 compared with the normal group; *p<0.05, **p<0.01 compared with the model group, (n=10);

[0079] Figure 7 The liver phenotype and liver index test result diagram in Example 2 of the present invention, wherein A is a liver morphology diagram, B is a liver index test result diagram, ####p<0.0001 compared with the normal group; *p<0.05 compared with the model group, (n=10);

[0080] Figure 8The results of alcohol detoxification-related enzyme detection in Example 2 of the present invention are shown in Figure 2, wherein A is the results of ADH enzyme detection, and B is the results of ALDH enzyme detection, ##p<0.01, ###p<0.001 compared with the normal group; *p<0.05, **p<0.01 compared with the model group, (n=10);

[0081] Fig. 9 : It is a test result diagram of mouse serum biochemical indicators in Example 2 of the present invention, wherein A is a test result diagram of AST, B is a test result diagram of ALT, C is a test result diagram of TC, D is a test result diagram of TG, and E is a test result diagram of LDL-C; F is a test result diagram of HDL-C, ##p<0.01, ###p<0.001, ####p<0.0001 compared with the normal group; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 compared with the model group, (n=10);

[0082] Fig.10 It is the test result diagram of the biochemical indicators in Example 2 of the present invention, wherein A is the CAT test result diagram, B is the GSH test result diagram, C is the MDA test result diagram, and D is the SOD test result diagram, ##p<0.01, ####p<0.0001 compared with the normal group; *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 compared with the model group, (n=10). DETAILED DESCRIPTION

[0083] The following will be combined with the embodiments to clearly and completely describe the concept of the present invention and the technical effects produced, so as to fully understand the purpose, characteristics and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative work are all within the scope of protection of the present invention.

[0084] Example 1 Preparation of Jasmine Flower Polysaccharide

[0085] This example prepares a frangipani polysaccharide, and the specific preparation and verification methods are as follows:

[0086] 1. Extraction of Jasmine Flower Polysaccharide

[0087] (1) crushing: crushing the fully dried jasmine flowers, sieving, and drying to obtain jasmine flower pollen;

[0088] (2) Degreasing: Jasmine pollen is degreased with 10 times the amount of anhydrous ethanol (i.e., the ratio of Jasmine pollen to anhydrous ethanol is 1 g: 10 mL) at 85°C and refluxed for 90 min. The reflux is repeated once, filtered, and the anhydrous ethanol is evaporated to obtain the residue, which is then dried.

[0089] (3) Ultrasonic-assisted extraction: The defatted dry powder was mixed with distilled water at a solid-liquid ratio of 1 g:15 mL, with an ultrasonic power of 100 W for 20 min, and then placed in a 100 °C water bath for extraction for 2.5 h. After centrifugation, the aqueous extract was filtered;

[0090] (4) Alcohol precipitation: The filtrate is concentrated to 20% of the original volume using a rotary evaporator, and then 4 times the amount of anhydrous ethanol is added to the concentrate, which is placed in a refrigerator overnight, and the precipitate is collected by filtration. The ethanol on the surface is evaporated in a fume hood to obtain a precipitate;

[0091] (5) Washing: The precipitate is washed twice with anhydrous ethanol, ether, and acetone in sequence, and dried to obtain a crude polysaccharide extract I;

[0092] (6) Protein removal: Take the above-mentioned crude polysaccharide extract I and dissolve it in an appropriate amount of water, add Sevag reagent according to the ratio of crude polysaccharide to Sevag reagent 5:1, shake thoroughly for 30 minutes, and then centrifuge at 6000r / min for 15 minutes; carefully aspirate the upper polysaccharide solution, remove the organic phase and precipitate, and then add Sevag solvent equivalent to 1 / 5 of its volume, repeat several times until no obvious precipitation is produced, and store at 4°C to prevent deterioration for subsequent impurity removal;

[0093] (7) Impurity removal: Use a ready-to-use dialysis bag of >8000D to remove impurities. First, cut the dialysis bag into a suitable length, soak the preservation solution with deionized water, place the deproteinized frangipani polysaccharide solution in the bag, seal it with a dialysis bag clip, and place a weight at the bottom to make it vertical to the bottom; then immerse the dialysis bag in a beaker filled with deionized water and a magnet, and then place the entire beaker on a magnetic stirrer to speed up the dialysis rate, thereby assembling a complete dialysis device. Dialysis is performed for 48 hours, and the liquid in the dialysis bag is taken out and centrifuged to obtain a precipitate. After freeze-drying the precipitate, a crude frangipani polysaccharide extract II (i.e., purified frangipani polysaccharide) is obtained.

[0094] 2. Determination of Jasmine Flower Polysaccharide Content

[0095] (1) Glucose standard curve drawing and polysaccharide determination method

[0096] Standard curve drawing: The purified frangipani polysaccharide was determined by the phenol-sulfuric acid method. Accurately weigh 100 mg of glucose standard into a 100 mL volumetric flask, dilute to 100 mL with distilled water, and prepare a 0.1 mg / mL glucose standard solution; pipette the prepared glucose standard solution and dilute to 5 mL with distilled water, and prepare 0.02 mg / mL, 0.04 mg / mL, 0.06 mg / mL, 0.08 mg / mL, 0.10 mg / mL, 0.12 mg / mL, and 0.14 mg / mL respectively. Take 200 μL of each dilution and place them in 2 mL test tubes. Add 100 μL of 6% phenol and 500 μL of concentrated sulfuric acid to each test tube, let it stand for 10 min, shake it well, and leave it at room temperature for 20 min. Punch 200 μL into each well of a 96-well plate, and punch 3 wells into each test tube. Use an enzyme reader to measure the absorbance at a wavelength of 490 nm, and use distilled water as a blank control to obtain a standard curve (such as Figure 1 shown).

[0097] Determination of polysaccharide content: Take the total polysaccharide of Jasmine flower and prepare it into a 1mg / mL solution with distilled water, dilute it 10 times with distilled water, take 200μL with a pipette, put it into a 96-well plate, make 3 holes, add phenol and sulfuric acid in proportion, let it stand for 10 minutes, shake it well, put it at room temperature for 20 minutes, and measure the absorbance at a wavelength of 490nm with an enzyme marker. Calculate according to the following formula: Polysaccharide content (%) = (C×V2×D) / (W×V1×10)×100% (Formula 2); where C—sugar content calculated by the standard curve (mg); V2—total volume of the extract (mL); V1—volume taken for determination (mL); D—dilution factor; W—sample weight (g).

[0098] The standard curve of glucose measured by phenol-sulfuric acid method is as follows: Figure 1 As shown, the regression equation is y = 6.8588x-0.0177; the fit of the equation is R 2 =0.999. In the range of 0-0.14 mg / mL, there is a good linear relationship between glucose concentration and absorbance value.

[0099] The absorbance of total polysaccharide was measured, substituted into the regression equation to obtain the concentration of polysaccharide, and then substituted into (Formula 2) to obtain the total polysaccharide content of 66.03%. There are many factors that affect the concentration and purity of polysaccharides, and the main factors include the following: higher extraction temperature will cause polysaccharide decomposition, and more times of deproteinization by Sevage method will also cause polysaccharide loss.

[0100] 3. Ultraviolet spectrum analysis of frangipani polysaccharide

[0101] The purified Jasmine flower polysaccharide was dissolved in ultrapure water to prepare a polysaccharide solution with a concentration of 1 mg / mL, and the ultraviolet absorption spectrum of the polysaccharide solution was scanned (200-600 nm) using an ultraviolet visible spectrophotometer.

[0102] The results are as follows Figure 2 As shown in the figure, it can be seen that aromatic amino acids in proteins (such as tryptophan, tyrosine, and phenylalanine) have characteristic absorption peaks near 280nm, and frangipani polysaccharide has a small absorption peak near 280nm, indicating that it contains a small amount of protein; chlorophyll, carotenoids, etc. usually have characteristic absorption peaks near 380nm, and frangipani polysaccharide has no absorption peak at 380nm, indicating that it does not contain these pigments or the pigment content is extremely low.

[0103] 4. Infrared spectroscopy analysis of frangipani polysaccharides

[0104] Take a certain amount of the purified dried frangipani polysaccharide sample, mix it with dry KBr powder and press it into tablets. -1 Infrared spectrum scanning is carried out within the range and infrared spectrum graph is recorded.

[0105] The results are as follows Figure 3 As shown in the figure, it can be seen that the frangipani polysaccharide is at 3414.351cm -1 It is caused by the stretching vibration of OH, a strong absorption peak, which is a characteristic peak of sugars; it proves that the substance is sugar. -1 The absorption peak at wave number 1748.156 cm is the characteristic peak of CH stretching vibration; -1 It is the characteristic absorption peak of carboxyl group, indicating that frangipani polysaccharide contains carboxyl group. -1 The observed absorption peaks are related to the stretching of COC and COH, indicating the possible presence of a pyranose ring.

[0106] 5. Determination of relative molecular weight of frangipani polysaccharide

[0107] The molecular weight of the purified frangipani polysaccharide was determined by high performance gel permeation chromatography (HPGPC). The chromatographic conditions were as follows: mobile phase: 0.05 M NaCl solution; chromatographic column: BRT105-103-101 series gel column (8×300 mm); flow rate: 0.7 mL / min; column temperature: 40°C; injection volume: 25 μL; detector: differential detector RID-20A; analysis time: 70 min.

[0108] The molecular weight spectrum of Jasmine polysaccharide is as follows Figure 4As shown in the figure, it can be seen that the frangipani polysaccharide peaks at 34.879 min, its relative peak area is 100%, and its molecular weight is 69445 Da.

[0109] 6. Determination of monosaccharide composition of Jasmine polysaccharide

[0110] Use ion chromatography to determine the monosaccharide composition. Accurately weigh 5 mg of the purified frangipani polysaccharide sample and place it in an ampoule, add 2 mL of 3MTFA, and hydrolyze at 120°C for 3 h. Accurately transfer the acid hydrolysis solution to a tube and blow it dry with nitrogen, add 5 mL of water and vortex to mix. Pipette 100 μL and add 900 μL of deionized water, centrifuge at 12000 rpm for 5 min. Take the supernatant for IC analysis.

[0111] The results are shown in Table 1 and Figure 5 As shown, it can be seen that the main monosaccharide composition and molar percentage of Jasmine polysaccharide are rhamnose (Rha): arabinose (Ara): galactose (Gal): glucose (Glc): xylose (Xyl): mannose (Man): galacturonic acid (GalA) = 0.019: 0.109: 0.174: 0.220: 0.021: 0.022: 0.434; among which the monosaccharides with the highest content are galacturonic acid (GalA), glucose (Glc), galactose (Gal) and arabinose (Ara).

[0112] Table 1

[0113]

[0114] Example 2 Application of Jasmine Flower Polysaccharide in the Preparation of Products for Sobering Up and Protecting the Liver

[0115] In this example, an ethanol-induced acute alcoholic liver injury model in mice was constructed to verify whether Jasmine polysaccharide has the effect of sobering up and protecting the liver. The specific verification method is as follows:

[0116] 1. Animals and groups

[0117] Animal model: 70 commercially available SPF grade C57BL / 6 male mice, aged 6-8 weeks, weighing 20-24g; after one week of adaptive feeding, the mice were randomly divided into 7 groups, 10 mice in each group, and the average weight of each group was about 23g. The specific groups were: (1) normal group (NC); (2) model group (Alcohol); (3) silymarin group (SM, 50mg / kg·BW); (4) King Drink group (King Drink, 150mg / kg·BW); (5) low-dose jasmine polysaccharide group (SP-L, 50mg / kg·BW); (6) medium-dose jasmine polysaccharide group (SP-M, 100mg / kg·BW); (7) high-dose jasmine polysaccharide group (SP-H, 300mg / kg BW).

[0118] 2. Experimental methods

[0119] (1) Construction of a 50% ethanol-induced acute alcoholic liver injury model in C57BL / 6 mice

[0120] The mice were given oral administration at 0.1 ml / 10 g for 14 days, and the body weight was recorded every two days. The normal group and the model group were given oral administration of pure water, and the other groups were given oral administration according to the above dosage. 2 hours after the last administration, except for the normal group who were given oral administration of pure water, the other mice were given oral administration of 50% ethanol at 0.14 ml / 10 g to create an acute alcoholic liver injury model.

[0121] After 30 minutes, the drunkenness time and soberness time of the mice were recorded, and the drunkenness rate of the mice after modeling was recorded (drunkenness rate = number of drunk mice / total number of mice × 100%). The drunkenness of the mice was determined by the disappearance of the righting reflex. After gavage, the mice were placed with their backs down and gently placed in a cage. If the mice maintained the back-down posture for more than 30 seconds, it was considered that the righting reflex disappeared, indicating that they were drunk. If the righting reflex of the drunken mice recovered and they could move freely, they were sober.

[0122] After excluding mice that died immediately after oral gavage and mice that did not show drunkenness for more than 3 hours, the number of drunk mice, drunkenness time and soberness time of each group of mice were observed and recorded.

[0123] After gavage with 50% ethanol, the mice were weighed after fasting for 16 hours but not water. Then the eyeballs were removed and blood was collected and placed in a 1.5 ml ordinary centrifuge tube at room temperature for more than 2 hours, centrifuged at 3000 rpm and 4°C for 10 minutes, and the supernatant was placed in another new centrifuge tube as serum, which was stored at -80°C for later use. After blood collection, the mice were killed, the liver and spleen were removed, washed in physiological saline and the excess blood was blotted out, the liver morphology was photographed, and the liver and spleen were weighed for the calculation of the organ coefficient: a part of the liver tissue was fixed in a 4% paraformaldehyde solution, and the rest was placed in a cryovial and quickly frozen with liquid nitrogen and stored at -80°C. It is used for the calculation of the liver index (liver index = liver (mg) / mouse body weight (mg) × 100%).

[0124] (2) Test results

[0125] 1) Results of mouse weight and intoxication rate test after modeling

[0126] The results are as follows Figure 6 As shown, from Figure 6 As can be seen from the weight graph A in the figure, the weight of mice in each group gradually increased during the 14 days of drug administration before modeling, indicating that Jasmine polysaccharide has no obvious toxicity to mice. After gavage with 50% ethanol for modeling, all groups of mice were fasted for 16 hours without water restriction. Compared with the NC group, the weight of mice in each drug administration group decreased to varying degrees, among which the weight loss of mice in the Alcohol group was the largest.

[0127] from Figure 6 As shown in the drunkenness rate test result chart B, compared with the normal group, the drunkenness rate of the model group increased significantly, indicating that the model was successfully established. The drunkenness rates of mice after intervention with Jasmine flower polysaccharide were reduced to varying degrees, among which the number of mice with drunkenness in the high-dose Jasmine flower polysaccharide group was the least, indicating that supplementing with Jasmine flower polysaccharide can reduce the drunkenness of mice.

[0128] 2) Liver phenotype and liver index

[0129] The results are as follows Figure 7 As shown, from Figure 7 As can be seen in Figure A, in the liver morphology images, the livers of mice in the normal group were ruddy and shiny, while the livers of mice in the alcohol group were enlarged, white, and had obvious granularity on the surface. The whitening of the liver surface of mice pretreated with frangipani polysaccharide was alleviated, and the granularity of the liver was weakened.

[0130] from Figure 7 As can be seen from Figure B, compared with the normal group, the liver index of the model group was significantly increased (p<0.0001), while the liver index of the Jasmine flower polysaccharide-treated group decreased in a dose-dependent manner.

[0131] 3) Detection of alcohol detoxification-related enzymes.

[0132] Experimental method: Cut 20-50 mg of liver tissue and place it in a 1.5 ml centrifuge tube. Add the homogenization medium extract at a ratio of 1:9 between tissue and normal saline. Use a manual tissue homogenizer to prepare 10% liver homogenate. Measure the activity of ADH and ALDH enzymes in turn according to the operating instructions of the kit.

[0133] The results are as follows Figure 8 As shown in the figure, it can be seen that compared with the normal group, the activities of ADH and ALDH enzymes in the model group were significantly reduced (p<0.01, p<0.0001). After treatment with jasmine flower polysaccharide, the activities of ADH and ALDH enzymes were significantly restored, and the effect was dose-dependent, indicating that jasmine flower polysaccharide can increase the activities of ADH and ALDH enzymes and accelerate the metabolism of ethanol in mice.

[0134] 4) Detection of mouse serum biochemical indicators

[0135] Experimental method: Mouse serum was obtained and diluted with normal saline for a certain multiple before direct testing; AST, ALT, TC, TG, LDL-C and HDL-C were determined according to the operating instructions of the Nanjing Jiancheng test kit.

[0136] The results are as follows Fig. 9 As shown in the figure, compared with the normal group, the model group mice significantly increased the content of AST, ALT, TC, TG and LDL-C in serum and reduced the content of HDL-C; while the ALT, AST, TC, TG and LDL-C content in the mice in the frangipani polysaccharide supplementation group were significantly reduced; the HDL-C content increased. This shows that frangipani polysaccharide can improve ethanol-induced acute alcoholic liver injury and abnormal lipid accumulation in mice.

[0137] 5) Detection of biochemical indices in mouse liver

[0138] Experimental method: Cut 20-50 mg of liver tissue and place it in a 1.5 ml centrifuge tube. Add normal saline as a homogenization medium at a ratio of 1:9 between tissue and normal saline. Use a manual tissue homogenizer to prepare 10% liver homogenate. Dilute it a certain number of times with normal saline to obtain the optimal detection concentration, and then use it for the detection of biochemical indicators. According to the operating instructions of the kit, the enzyme activity levels of CAT, SOD, GSH and the content of MDA are measured in sequence.

[0139] The results are as follows Fig.10As shown in the figure, compared with the normal group, the levels of CAT, SOD, and GSH in the liver of the model group were significantly reduced (p<0.01), and the level of MDA was significantly increased (p<0.0001). However, treatment with frangipani polysaccharide can significantly increase the levels of CAT, SOD, and GSH enzyme activities and reduce the content of MDA. These results show that frangipani polysaccharide can protect the liver from damage by enhancing antioxidant activity.

[0140] Jasmine is a winding vine-like evergreen ornamental shrub of the genus Jasmine (Oleaceae). Jasmine is bitter, mild and non-toxic, and belongs to the liver meridian; it has the traditional efficacy of "soothing the liver and relieving depression, promoting qi and relieving pain". Modern medicinal chemistry research has found that Jasmine contains iridoids, triterpenes, flavonoids, lignans and other ingredients. Among them, there are monomer compounds such as oleuropein, hydroxytyrosol, hydroquinone, kaempferol, ethyl gallate, salidroside and quercetin. It is a dual-purpose medicinal and edible resource rich in active ingredients. However, Jasmine polysaccharides and related activities have not been reported so far. The present invention is the first to find that Jasmine polysaccharides have the effect of sobering up, and can also improve ethanol-induced acute alcoholic liver injury.

[0141] The embodiments of the present invention are described in detail above in conjunction with the accompanying drawings, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. Application of Jasmine flower polysaccharide in the preparation of hangover relieving products.

2. Application of Jasmine flower polysaccharide in the preparation of products for preventing and / or treating liver damage caused by drinking.

3. The use according to claim 1 or 2, characterized in that: The product has at least one of the following functions: (1) Accelerate the metabolism of ethanol and / or acetaldehyde; (2) shorten the duration of intoxication and / or sobriety; (3) Reduce the rate of drunkenness; (4) Reduce liver index; (5) Increase the activity of ADH and / or ALDH enzymes in the body; (6) reduce serum ALT and / or AST levels; (7) Increase serum HDL-C levels; (8) Increase CAT, SOD and / or GSH activity; (9) Reduce MDA content; (10) Improve ethanol-induced abnormal lipid accumulation; (11) Prevent alcohol metabolism disorders caused by excessive alcohol intake; Preferably, the improvement of ethanol-induced abnormal lipid accumulation includes reducing the levels of triglycerides, cholesterol, and low-density lipoprotein cholesterol in serum.

4. The use according to any one of claims 1 to 3, characterized in that: The preparation method of the frangipani polysaccharide comprises the following steps: The frangipani flower sample is subjected to defatting, water extraction, alcohol precipitation, protein removal and dialysis in sequence to obtain frangipani flower polysaccharide; Preferably, the defatting is carried out using anhydrous ethanol; More preferably, in the defatting step, the solid-liquid ratio of the frangipani sample to anhydrous ethanol is 1 g: 8-12 mL; Preferably, the dialysis is performed using a dialysis bag with a molecular weight cut-off of >8000Da; Preferably, the dialysis time is 1-3 days.

5. The use according to any one of claims 1 to 3, characterized in that: The monosaccharides in the frangipani polysaccharide are composed of rhamnose, arabinose, galactose, glucose, xylose, mannose and galacturonic acid.

6. The use according to claim 5, characterized in that: The molar ratio of rhamnose, arabinose, galactose, glucose, xylose, mannose and galacturonic acid is (14-22): (100-120): (160-185): (200-240): (16-25): (18-27): (400-450).

7. The use according to any one of claims 1 to 3, characterized in that: The molecular weight of the frangipani polysaccharide is 69445 Da.

8. A product for sobering up or preventing liver damage caused by drinking, characterized in that: The product includes frangipani polysaccharide.

9. The product according to claim 8, characterized in that The product is at least one of a medicine, a health product and a functional food.

10. The product according to claim 8, characterized in that The product is a medicine, and the medicine also includes pharmaceutically acceptable excipients; Preferably, the dosage form of the drug is tablets, capsules, soft capsules, granules, pills, oral liquids, dry suspensions, pellets, dry extracts, injections or infusions, transdermal agents, and transdermal microneedles.

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