Composition with uric acid reducing effect and preparation method thereof

Abstract

The invention relates to the technical field of health care products, in particular to a composition with a uric acid reducing effect and a preparation method thereof. The composition with the uric acid reducing effect is prepared from the following components in parts by mass: 5 to 10 parts of N-acetylglucosamine, 2 to 5 parts of honeysuckle flower polysaccharide and 1 to 3 parts of radix pseudostellariae oligosaccharide. The composition provided by the invention has the effects of inhibiting xanthine oxidase, resisting inflammation, regulating intestinal flora, improving immunity, inhibiting creatinine level rise, relieving oxidative damage, reducing lipid level and the like, so that generation of uric acid is inhibited and reduced, and chronic damage to kidneys, cardiovascular systems and the like is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of health products, and particularly relates to a composition with the effect of reducing uric acid and a preparation method thereof. Background Art

[0002] Hyperuricemia is a common metabolic disease, a metabolic disease caused by purine metabolism disorder or reduced uric acid excretion. In recent years, with the improvement of people's living standards and the change of diet structure, the incidence rate has shown a significant upward trend. Long-term hyperuricemia will not only cause acute symptoms such as gouty arthritis and gouty tophus deposition, but also cause chronic damage to the kidneys, cardiovascular system, etc., increasing the risk of diseases such as renal failure, hypertension, and coronary heart disease, greatly increasing the health risks of patients and the social medical burden.

[0003] Currently, the clinical treatment of hyperuricemia mainly relies on drugs, but drug treatment has many limitations and may produce side effects when taken for a long time. For example, drugs that inhibit uric acid production (such as allopurinol and febuxostat) and drugs that promote uric acid excretion (such as benzbromarone). These drugs have side effects to varying degrees. Allopurinol may cause severe hypersensitivity reactions, febuxostat has cardiovascular risks, and benzbromarone may cause liver damage, etc. In contrast, health foods have the advantages of safety, convenience, and can be taken for a long time, and can meet the daily health care needs of the majority of hyperuricemia patients. Developing a safe and effective treatment composition for hyperuricemia is of great significance for improving the health status of hyperuricemia patients and improving the quality of life.

[0004] The pathogenesis of hyperuricemia mainly involves two aspects: excessive uric acid production and reduced excretion:

[0005] 1. Excessive uric acid production. Such as abnormal purine metabolism: Purine metabolism disorder will lead to increased uric acid production. Common causes include: Diet: Intake of too much high-purine food (such as red meat, seafood). Enzyme deficiency: Such as the lack of hypoxanthine-guanine phosphoribosyltransferase (HGPRT), resulting in abnormal purine metabolism. Accelerated cell decomposition: Such as diseases such as tumors and hemolytic anemia leading to massive cell decomposition and release of purines.

[0006] 2. Reduced uric acid excretion. Such as abnormal kidney function: Uric acid is mainly excreted through the kidneys, and decreased kidney function will reduce uric acid excretion. Common causes are: Reduced glomerular filtration rate: Such as chronic kidney disease. Increased tubular reabsorption: Such as certain drugs (diuretics, aspirin) affecting tubular function. Genetic factors: Such as gene mutations of transporters such as URAT1, affecting uric acid excretion.

[0007] 3. Other factors, such as metabolic syndrome: Metabolic abnormalities such as obesity, hypertension, and diabetes are often accompanied by hyperuricemia. Drugs: Certain drugs (such as diuretics and cyclosporine) may interfere with uric acid metabolism. Genetic factors: Some people are more susceptible to the disease due to genetic susceptibility.

[0008] 4. Pathophysiological mechanisms, such as uric acid crystal deposition: When the blood uric acid level is too high, uric acid crystals may deposit in joints, kidneys and other parts, causing complications such as gout and kidney stones. Inflammatory response: Uric acid crystals can activate inflammatory pathways, leading to local inflammation and tissue damage.

[0009] Therefore, the pathogenesis of hyperuricemia is complex, involving the combined effects of excessive uric acid production, reduced excretion and many other factors. The treatment targets of hyperuricemia mainly focus on metabolic links such as uric acid production, excretion and reabsorption, as well as pathological processes such as inflammation and oxidative stress related to hyperuricemia, but the treatment effects are not satisfactory.

[0010] N-acetylglucosamine (NAG) is an amino monosaccharide that plays a key role in living organisms. As the basic constituent unit of chitin, the second largest natural polysaccharide in nature, it is widely present in microorganisms such as fungi and bacteria, as well as in animal and plant cells, especially in human connective tissues, articular cartilage, skin and intestinal mucosa. In terms of human physiological functions, NAG plays a crucial role. It is not only an important prerequisite for the synthesis of bifidogenic factors, but also a donor of N-acetylglucosamine residues for glycosaminoglycans, proteoglycans, lipopolysaccharides, etc., which is essential for maintaining the normal physiological functions of organisms. At present, NAG has been widely used in the fields of bone and joint health and skin. NAG has significant anti-inflammatory, antioxidant and metabolic regulatory effects. Based on this, how to develop more functions of NAG and apply it in the treatment of hyperuricemia is an urgent problem for those skilled in the art. Summary of the Invention

[0011] The object of the present invention is to provide a composition with uric acid-lowering efficacy and a preparation method thereof.

[0012] In order to achieve the above-mentioned invention object, the present invention provides the following technical solutions:

[0013] The present invention provides the use of N-acetylglucosamine in the preparation of a health product with uric acid-lowering efficacy.

[0014] The present invention also provides a composition with uric acid-lowering efficacy, comprising the following components in parts by mass: 5-10 parts of N-acetylglucosamine, 2-5 parts of honeysuckle polysaccharide, and 1-3 parts of heterophylly falsestarwort oligosaccharide.

[0015] Preferably, the preparation method of the honeysuckle polysaccharide is as follows: reflux extract honeysuckle with ethanol to obtain an extract; concentrate the extract and then perform alcohol precipitation to obtain a precipitate, which is the crude honeysuckle polysaccharide; wash the crude honeysuckle polysaccharide with Sevag reagent and pass it through a cellulose chromatography column to obtain the honeysuckle polysaccharide.

[0016] Preferably, the method of reflux extraction with ethanol is as follows: reflux extract according to honeysuckle: 95% ethanol = 1 g: 5 - 20 ml for 1 - 2 times, 1 - 2 h each time. After filtration, reflux extract the residue: 50% ethanol = 1 g: 5 - 20 ml for 2 - 3 times, 1 - 2 h each time.

[0017] Preferably, the alcohol precipitation is as follows: add 4 - 5 volumes of absolute ethanol to the concentrated extract and let it stand at 4 - 6 °C for 12 - 16 h; the number of times of washing is 8 - 12 times.

[0018] Preferably, the preparation method of the pseudostellaria heterophylla oligosaccharide is as follows: reflux extract pseudostellaria heterophylla with ethanol to obtain an extract; concentrate the extract, then perform extraction and alcohol precipitation, and take the supernatant; concentrate the supernatant and pass it through a PA20 gel column for enrichment. According to the HPLC detection results, collect the part with a degree of polymerization of 2 - 10 to obtain the pseudostellaria heterophylla oligosaccharide.

[0019] Preferably, the method of reflux extraction with ethanol is as follows: reflux extract according to honeysuckle: 50% ethanol = 1 g: 5 - 20 ml for 2 - 3 times, 1 - 2 h each time.

[0020] Preferably, the extractant used for extraction is ethyl acetate; the volume ratio of the concentrated extract to ethyl acetate during extraction is 1:5 - 20; the number of times of extraction is 3 - 4 times; the alcohol precipitation is as follows: add 4 - 5 volumes of absolute ethanol to the extract after extraction and let it stand at 4 - 6 °C for 12 - 16 h to obtain the supernatant.

[0021] The present invention also provides a preparation method of the composition, which is to mix N - acetylglucosamine, honeysuckle polysaccharide, pseudostellaria heterophylla oligosaccharide and water at 40 - 60 °C, and dissolve them by ultrasonic wave to prepare a drug solution with a concentration of 40 - 60 mg / mL.

[0022] The present invention also provides the application of the composition or the composition prepared by the preparation method in the preparation of health products with the efficacy of reducing uric acid.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] Through the compound synergism of N-acetylglucosamine, honeysuckle polysaccharide and heterophylly falsestarwort root oligosaccharide, the composition has the effects of inhibiting xanthine oxidase, anti-inflammation, regulating intestinal flora, enhancing immunity, inhibiting the increase of creatinine level, reducing oxidative damage, and lowering lipid level, etc., thereby inhibiting and reducing the generation of uric acid and reducing the chronic damage to the kidneys, cardiovascular system, etc., and having the technical effect of safely and effectively reducing uric acid. Brief Description of the Drawings

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained according to the provided drawings without creative efforts.

[0026] Figure 1 Changes in serum uric acid levels of mice in different groups (* indicates significant difference compared with the blank group, *p < 0.05, ***p < 0.001, ****p < 0.0001; # indicates significant difference compared with the model group, #p < 0.05, ##p < 0.01, p < 0.001, #p < 0.0001; the same below).

[0027] Figure 2 Results of the determination of liver XOD activity of mice in different groups.

[0028] Figure 3 Results of the determination of creatinine levels of mice in different groups.

[0029] Figure 4 Results of the determination of inflammatory factor IL-6 in the serum of mice in different groups.

[0030] Figure 5 Results of the determination of inflammatory factor IL-1β in the serum of mice in different groups.

[0031] Figure 6 Results of the determination of inflammatory factor TNF-α in the serum of mice in different groups.

[0032] Figure 7 Results of the determination of MDA levels of mice in different groups.

[0033] Figure 8 Results of the determination of total cholesterol (TCHO) levels of mice in different groups. Detailed Embodiments

[0034] The present invention provides the application of N-acetylglucosamine in the preparation of health products with the efficacy of reducing uric acid.

[0035] The present invention also provides a composition with the efficacy of reducing uric acid, comprising the following components in parts by mass: 5-10 parts of N-acetylglucosamine, 2-5 parts of honeysuckle polysaccharide, and 1-3 parts of pseudostellaria heterophylla oligosaccharide; preferably 6-9 parts of N-acetylglucosamine, 3-4 parts of honeysuckle polysaccharide, and 2 parts of pseudostellaria heterophylla oligosaccharide; more preferably 7-8 parts of N-acetylglucosamine, 4 parts of honeysuckle polysaccharide, and 2 parts of pseudostellaria heterophylla oligosaccharide; even more preferably 8 parts of N-acetylglucosamine, 4 parts of honeysuckle polysaccharide, and 2 parts of pseudostellaria heterophylla oligosaccharide.

[0036] In the present invention, the preparation method of the honeysuckle polysaccharide is as follows: Honeysuckle is refluxed and extracted with ethanol to obtain an extract; the extract is concentrated and then precipitated with alcohol to obtain a precipitate, which is the crude honeysuckle polysaccharide; the crude honeysuckle polysaccharide is washed with Sevag reagent and passed through a cellulose chromatography column to obtain the honeysuckle polysaccharide.

[0037] In the present invention, the method of ethanol reflux extraction is as follows: Honeysuckle: 95% ethanol = 1 g: 5-20 ml is refluxed and extracted 1-2 times, each time for 1-2 h. After filtration, the residue: 50% ethanol = 1 g: 5-20 ml is refluxed and extracted 2-3 times, each time for 1-2 h;

[0038] Preferably, honeysuckle: 95% ethanol = 1 g: 10-15 ml is refluxed and extracted 2 times, each time for 2 h. After filtration, the residue: 50% ethanol = 1 g: 10-15 ml is refluxed and extracted 3 times, each time for 2 h; more preferably, honeysuckle: 95% ethanol = 1 g: 13 ml is refluxed and extracted 2 times, each time for 2 h. After filtration, the residue: 50% ethanol = 1 g: 13 ml is refluxed and extracted 3 times, each time for 2 h.

[0039] In the present invention, the alcohol precipitation is as follows: The concentrated extract is added with 4-5 volumes of absolute ethanol and left to stand at 4-6 °C for 12-16 h; preferably, the concentrated extract is added with 5 volumes of absolute ethanol and left to stand at 4 °C for 13-15 h; more preferably, the concentrated extract is added with 5 volumes of absolute ethanol and left to stand at 4 °C for 14 h.

[0040] In the present invention, the number of times of washing is 8-12 times; preferably 9-11 times; more preferably 10 times.

[0041] In the present invention, the preparation method of the pseudostellaria heterophylla oligosaccharide is as follows: Pseudostellaria heterophylla is refluxed and extracted with ethanol to obtain an extract; the extract is concentrated and then extracted, precipitated with alcohol, and the supernatant is taken; the supernatant is concentrated and passed through a PA20 gel column for enrichment. According to the detection results of HPLC, the fraction with a degree of polymerization of 2-10 is collected to obtain the pseudostellaria heterophylla oligosaccharide.

[0042] In the present invention, the method of ethanol reflux extraction is as follows: Flos Lonicerae: 50% ethanol = 1 g: 5 - 20 ml, reflux extraction is carried out 2 - 3 times, 1 - 2 h each time; preferably, Flos Lonicerae: 50% ethanol = 1 g: 10 - 15 ml, reflux extraction is carried out 3 times, 2 h each time; more preferably, 50% ethanol = 1 g: 13 ml, reflux extraction is carried out 3 times, 2 h each time.

[0043] In the present invention, the extractant used for extraction is ethyl acetate; when extracting, the volume ratio of the concentrated extract to ethyl acetate is 1: 5 - 20; preferably 1: 10 - 15; more preferably 1: 13.

[0044] In the present invention, the number of extraction times is 3 - 4 times; preferably 3 times.

[0045] In the present invention, alcohol precipitation is as follows: Add 4 - 5 times the volume of absolute ethanol to the extract after extraction, stand at 4 - 6 °C for 12 - 16 h to obtain the supernatant; preferably, add 5 times the volume of absolute ethanol to the extract after extraction, stand at 4 °C for 13 - 15 h to obtain the supernatant; more preferably, add 5 times the volume of absolute ethanol to the extract after extraction, stand at 4 °C for 14 h to obtain the supernatant.

[0046] The present invention also provides a preparation method of the composition. Mix N - acetylglucosamine, Flos Lonicerae polysaccharide, Radix Pseudostellariae oligosaccharide and water at 40 - 60 °C, dissolve by ultrasonic wave, and prepare a drug solution with a concentration of 40 - 60 mg / mL; preferably 45 - 55 mg / mL; more preferably 50 mg / mL.

[0047] The present invention also provides the application of the composition or the composition prepared by the preparation method in the preparation of a health product with the efficacy of reducing uric acid.

[0048] The following is a detailed description of the technical solutions provided by the present invention in conjunction with the examples, but they cannot be understood as limiting the protection scope of the present invention.

[0049] Example 1

[0050] A preparation method of a composition with the efficacy of reducing uric acid is as follows:

[0051] (1) Preparation of Flos Lonicerae polysaccharide:

[0052] 1kg of honeysuckle medicinal material was extracted twice with 15 times 95% ethanol by heating and refluxing, each time for 2 hours. The filtered residue was extracted twice with 15 times 50% ethanol by heating and refluxing, each time for 2 hours. The extracts were mixed and concentrated under reduced pressure to obtain 500mL of extract, and 5 times the volume of anhydrous ethanol was added. The extract was left to stand at 4°C overnight to precipitate and obtain crude honeysuckle polysaccharide. The crude polysaccharide was dissolved in water, Sevag reagent was added, and the extract was washed repeatedly for 10 times to remove the protein to obtain crude honeysuckle polysaccharide. The crude honeysuckle polysaccharide was further purified by cellulose chromatography column, and eluted with ultrapure water, 0.1, 0.3, and 0.5 mol / L NaCl aqueous solution at a flow rate of 1 mL / min, and a fraction was collected every 5 mL. The polysaccharide content was determined by phenol-sulfuric acid method, and the fractions containing honeysuckle polysaccharide were combined and loaded into dialysis bags, dialyzed at 4°C for 48 hours to remove NaCl (the same below), and honeysuckle polysaccharide was enriched.

[0053] (2) Preparation of Pseudostellariae oligosaccharide:

[0054] 1 kg of Radix Pseudostellariae Radix was extracted twice with 15 times 50% ethanol by heating and refluxing, each time for 2 hours. The extracts were mixed and concentrated to 300 mL under reduced pressure, extracted three times with an equal volume of ethyl acetate to remove fat-soluble components, 5 times the volume of anhydrous ethanol was added to the aqueous phase, polysaccharides and proteins were precipitated and removed, and the supernatant was concentrated and enriched by a PA20 gel column, ultrapure water was used as the eluent, and a fraction was collected every 5 mL. The collected fractions were analyzed by HPLC using an amino column, an evaporative light scattering detector, and the HPLC conditions were: 0-30 min, 75-55% acetonitrile, 30-32 min, 55-10% acetonitrile, 32-35 min, 10% acetonitrile (the same below). According to the HPLC test results, the part with a degree of polymerization of 2 to 10 was collected to obtain Radix Pseudostellariae Radix oligosaccharides.

[0055] (3) The raw material composition of composition 1 is: 10 parts of N-acetylglucosamine, 5 parts of honeysuckle polysaccharide, and 2 parts of Pseudostellariae oligosaccharide, and the parts are all by mass.

[0056] (4) The preparation method is as follows: N-acetylglucosamine, honeysuckle polysaccharide, and Pseudostellaria oligosaccharide are mixed in a mass ratio of 10:5:2, added with water at a temperature of 40°C, dissolved by ultrasonic, and prepared into a 50 mg / mL solution of composition 1, which is stored at 4°C for later use.

[0057] Example 2

[0058] A method for preparing a composition having the effect of lowering uric acid, comprising the following steps:

[0059] (1) Preparation of honeysuckle polysaccharide:

[0060] 1 kg of Lonicera japonica Thunb. medicinal materials were extracted by heating under reflux with 15 times the volume of 95% ethanol twice, each time for 2 h. The residue after filtration was extracted by heating under reflux with 15 times the volume of 50% ethanol twice, each time for 2 h. The extraction solutions were mixed and concentrated under reduced pressure to obtain 500 mL of an extraction solution. Five times the volume of absolute ethanol was added, and the mixture was allowed to stand overnight at 4°C to precipitate crude Lonicera japonica Thunb. polysaccharide. The crude polysaccharide was dissolved in water, Sevag reagent was added, and it was washed repeatedly 10 times to remove proteins, obtaining a Lonicera japonica Thunb. polysaccharide solution. Through a cellulose chromatography column, the Lonicera japonica Thunb. polysaccharide was further enriched.

[0061] (2) Preparation of Pseudostellaria heterophylla oligosaccharide:

[0062] 1 kg of Pseudostellaria heterophylla medicinal materials were extracted by heating under reflux with 15 times the volume of 50% ethanol twice, each time for 2 h. The extraction solutions were mixed and concentrated under reduced pressure to 300 mL, and extracted three times with an equal volume of ethyl acetate to remove liposoluble components. Five times the volume of absolute ethanol was added to the aqueous phase to precipitate and remove polysaccharides and proteins. The supernatant was concentrated and enriched through a PA20 gel column. According to the HPLC detection results, the fraction with a degree of polymerization of 2 - 10 was collected to obtain Pseudostellaria heterophylla oligosaccharide.

[0063] (3) The raw material composition of Composition 2 is: 8 parts of N - acetylglucosamine, 2 parts of Lonicera japonica Thunb. polysaccharide, and 3 parts of Pseudostellaria heterophylla oligosaccharide, and the parts are all by mass.

[0064] (4) Preparation method: N - acetylglucosamine, Lonicera japonica Thunb. polysaccharide, and Pseudostellaria heterophylla oligosaccharide were mixed according to a mass ratio of 8:2:3, added to water at a temperature of 60°C, and dissolved by ultrasonic treatment to prepare a 50 mg / mL Composition 2 solution, which was stored for standby at 4°C.

[0065] Example 3

[0066] A preparation method of a composition with uric acid - lowering efficacy is as follows:

[0067] (1) Preparation of Lonicera japonica Thunb. polysaccharide:

[0068] 1 kg of Lonicera japonica Thunb. medicinal materials were extracted by heating under reflux with 15 times the volume of 95% ethanol twice, each time for 2 h. The residue after filtration was extracted by heating under reflux with 15 times the volume of 50% ethanol twice, each time for 2 h. The extraction solutions were mixed and concentrated under reduced pressure to obtain 500 mL of an extraction solution. Five times the volume of absolute ethanol was added, and the mixture was allowed to stand overnight at 4°C to precipitate crude Lonicera japonica Thunb. polysaccharide. The crude polysaccharide was dissolved in water, Sevag reagent was added, and it was washed repeatedly 10 times to remove proteins, obtaining a Lonicera japonica Thunb. polysaccharide solution. Through a cellulose chromatography column, the Lonicera japonica Thunb. polysaccharide was further enriched.

[0069] (2) Preparation of Pseudostellaria heterophylla oligosaccharide:

[0070] 1 kg of Pseudostellariae Radix was extracted twice by heating under reflux with 15 times its volume of 50% ethanol for 2 hours each time. The extracts were combined and concentrated under reduced pressure to 300 mL, then extracted 3 times with an equal volume of ethyl acetate to remove lipophilic components. Five times the volume of absolute ethanol was added to the aqueous phase to precipitate and remove polysaccharides and proteins. The supernatant was concentrated and enriched through a PA20 gel column. According to the HPLC test results, the fraction with a degree of polymerization of 2 - 10 was collected to obtain Pseudostellariae Radix oligosaccharides.

[0071] (3) The raw material composition of Composition 3 is: 5 parts of N - acetylglucosamine, 5 parts of Lonicera japonica polysaccharide, and 1 part of Pseudostellariae Radix oligosaccharides, and the parts are all in mass parts.

[0072] (4) The preparation method is: Mix N - acetylglucosamine, Lonicera japonica polysaccharide, and Pseudostellariae Radix oligosaccharides in a mass ratio of 5:5:1, add water at a temperature of 45°C, and dissolve by ultrasonic treatment to prepare a 50 mg / mL Composition 3 solution, which is stored for later use at 4°C.

[0073] Comparative Example 1

[0074] The technical solution of the uric acid - lowering composition is similar to that of Example 2, except that in the composition of Comparative Example 1, N - acetylglucosamine is not added, and glucose is added instead.

[0075] The preparation method is: Mix glucose, Lonicera japonica polysaccharide, and Pseudostellariae Radix oligosaccharides in a mass ratio of 8:2:3, add water at a temperature of 45°C, and dissolve by ultrasonic treatment to prepare a 50 mg / mL Composition 4 solution, which is stored for later use at 4°C.

[0076] Comparative Example 2

[0077] The technical solution of the uric acid - lowering composition is similar to that of Example 2, except that in the composition of Comparative Example 2, Lonicera japonica polysaccharide and Pseudostellariae Radix oligosaccharides are not added, and Forsythia suspensa and Codonopsis pilosula are added instead.

[0078] The preparation method is: Mix N - acetylglucosamine, Forsythia suspensa extract, and Codonopsis pilosula extract in a mass ratio of 5:3:2, add water at a temperature of 45°C, and dissolve by ultrasonic treatment to prepare a 50 mg / mL Composition 5 solution, which is stored for later use at 4°C.

[0079] Comparative Example 3

[0080] The technical solution of the uric acid - lowering composition is similar to that of Example 2, except that in the composition of Comparative Example 3, N - acetylglucosamine and Lonicera japonica polysaccharide are not added, and maltose and Platycodon grandiflorum are added instead.

[0081] The preparation method is as follows: maltose, platycodon grandiflorum extract and pseudostellaria heterophylla oligosaccharide are mixed according to a mass ratio of 6:3:5, water at 45 °C is added, and ultrasonic dissolution is carried out to prepare a composition 6 solution with a concentration of 50 mg / mL, which is stored for standby at 4 °C.

[0082] Experimental Example 1

[0083] 1 Animal grouping

[0084] 72 SPF-grade Kunming (KM) mice with a body weight of 23 - 25 g are randomly divided into 9 groups: blank group, model group, positive control group, Example 1 group, Example 2 group, Example 3 group, Comparative Example 1 group, Comparative Example 2 group, and Comparative Example 3 group (corresponding to blank, model, control, Example 1, Example 2, Example 3, Comparative 1, Comparative 2, and Comparative 3 groups respectively).

[0085] 2 Modeling method

[0086] An HUA mouse model is induced by intraperitoneal injection of PO solution (300 mg / kg, i.p. potassium oxonate) and intragastric administration of HX solution (300 mg / kg, i.g.) combined with feeding a 20% high-yeast diet. The mice are adaptively fed for one week, and then the induction and administration of the HUA mouse model are carried out. The mice in the blank group are given normal diet, while the mice in the other groups are all fed with a 20% high-yeast diet. Every morning at 8 o'clock, each mouse's body weight is weighed and recorded using a small weighing scale, and all modeling solutions, administration solutions, etc. are heated to 37 °C in a water bath for warming and standby.

[0087] The mice in the model group, positive drug group, example groups, and comparative example groups are all modeled by intraperitoneal injection of PO (300 mg / kg, i.p.) solution and intragastric administration of HX (hypoxanthine) solution (300 mg / kg, i.g.), while the mice in the blank group are given an equal amount of 0.5% CMC solution (i.p., i.g.).

[0088] 1.5 hours after modeling, the mice in the blank control group and the model group are intragastrically administered 0.9% normal saline daily, the positive drug group is intragastrically administered AP (allopurinol) solution (5 mg / kg, i.g.), and the administration groups are intragastrically administered solutions of compositions 1 - 6 respectively. The volumes of all modeling solutions and administration solutions are calculated according to 0.1 mL / 10 g and administered accordingly. After continuous administration for 14 days, samples are taken for detection.

[0089] 3 Collection of serum and tissue samples

[0090] On the 14th day after the last modeling and 1 hour after drug administration, mice were anesthetized by intraperitoneal injection of sodium pentobarbital solution, and then blood was collected from the eyeballs. The blood was placed in a 1.5 mL DNase / RNase-Free centrifuge tube and left to stand at room temperature for 1 hour. After 1 hour, the blood was centrifuged at 3500 rpm for 10 minutes, and then the supernatant was aliquoted into 1.5 mL DNase / RNase-Free centrifuge tubes and stored at -80 °C until further detection and analysis.

[0091] After blood collection from the eyeballs, the mice were quickly dissected, and the kidneys, livers, and small intestines were removed respectively. After gently washing the tissues in physiological saline, the water on the surface of the tissues was gently blotted with absorbent paper, and the weights of the kidneys and livers were weighed and recorded. The left kidneys and livers were placed in DNase / RNase-Free centrifuge tubes respectively, the centrifuge tube caps were covered and quickly placed in liquid nitrogen. The right kidneys and small intestines were placed in DNase / RNase-Free centrifuge tubes containing 4% paraformaldehyde for tissue fixation, which were used for later tissue H&E staining and IHC detection and analysis.

[0092] 4 Sample Detection

[0093] The experimental operation steps of all biochemical indexes were strictly carried out according to the kit instructions, and the levels of uric acid (UA), xanthine oxidase (XOD) activity, inflammatory factors TNF-α, IL-1β, IL-6 in mouse serum, and indexes related to kidney injury creatinine (CRE), malondialdehyde (MDA) content as an oxidative stress index, and total cholesterol (TCHO) level in small intestine were measured using an enzyme-linked immunosorbent assay (ELISA) reader.

[0094] Results:

[0095] 1 Uric Acid (UA) Level

[0096] Figure 1 The changes in serum uric acid levels of mice in different groups are shown. The serum uric acid level of mice in the model group was significantly higher than that in the blank control group, which verified the effectiveness of the model. The compositions in Examples 1 to 3 could all significantly reduce the uric acid level of mice, and the effect was comparable to that of the traditional uric acid-lowering drug allopurinol. In contrast, although Comparative Examples 1 to 3 could also reduce the serum uric acid level, the effect was not as good as that of Examples 1 to 3. The experimental results further showed that the absence of any component in the composition would weaken its effect of reducing uric acid.

[0097] 2 Xanthine Oxidase (XOD) Activity

[0098] Xanthine oxidase plays a crucial role in the production of uric acid. It catalyzes the conversion of hypoxanthine to xanthine and further converts xanthine to uric acid. Therefore, xanthine oxidase has a decisive impact on the production of uric acid. When the activity of xanthine oxidase increases, the production rate of uric acid will accelerate, which may lead to the occurrence or exacerbation of hyperuricemia. As Figure 2 shown, the hepatic XOD activity of the mice in the model group was significantly higher than that in the blank control group, indicating that the increased XOD activity in the state of hyperuricemia was successfully simulated by the modeling method; Implementations 1 to 3 could all significantly reduce the XOD activity of the mice, and the effect was similar to that of the positive drug allopurinol group, further confirming the potential of this composition in reducing XOD activity and thus reducing the blood uric acid level; while Comparisons 1 to 2 had less impact on XOD activity, and the impact of Comparison 3 on XOD activity had no significant difference compared with the model, indicating that the lack of any component would weaken the inhibitory effect of the composition on XOD activity, thereby reducing its effect on reducing blood uric acid.

[0099] 3 Creatinine (CRE) activity

[0100] As a biomarker of renal function, the level of creatinine will increase when renal function is impaired. The decline of renal function will lead to the reduction of uric acid excretion, which may trigger or exacerbate hyperuricemia. In clinical practice, doctors usually detect the levels of creatinine and uric acid simultaneously to evaluate the status of renal function and uric acid metabolism. As Figure 3 shown, compared with the control group, the creatinine level of the hyperuricemic mice was significantly increased, indicating that the kidneys of the mice in the model group were damaged to a certain extent. The positive control AP group failed to reduce the increased serum creatinine level caused by hyperuricemia, but instead increased. However, the administration groups of Implementations 1 to 3 significantly reduced the serum creatinine level compared with the model group. In contrast, the administration groups of Comparisons 1 to 3 did not show obvious differences compared with the model group. This indicates that the lack of any component will weaken the ability of the composition to inhibit the increase of serum creatinine level, thereby weakening its protective effect on renal function.

[0101] 4 Inflammatory factors TNF-α, IL-1β, IL-6

[0102] In hyperuricemia, the blood uric acid level increases, and urate crystals can be recognized by immune cells (such as macrophages, neutrophils, etc.). By activating inflammatory signaling pathways such as the NLRP3 inflammasome, it promotes the synthesis and release of inflammatory factors such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). These inflammatory factors will further amplify the inflammatory response, leading to local and systemic inflammatory states. Figures 4 - 6The measurement results of inflammatory factors IL-6, IL-1β and TNF-α in serum are shown. The results show that compared with the blank group, the contents of TNF-α, IL-1β and IL-6 in the serum of mice in the model group are significantly increased; compared with the model group, the levels of IL-1β and TNF-α in the positive drug AP group are significantly decreased, and the level of IL-6 also decreases. Compared with the model group, the administration groups of Examples 1 to 3 and Comparative Examples 1 to 3 can effectively reduce the contents of TNF-α, IL-1β and IL-6. However, compared with Comparative Examples 1 to 3, the differences between Examples 1 to 3 and the model group are more significant. This shows that if any component is missing in the composition, its inhibitory effect on the increase of inflammatory factors TNF-α, IL-1β and IL-6 will be weakened, and further the improvement effect on hyperuricemia and related complications will be weakened.

[0103] 5 MDA in liver tissue

[0104] MDA (malondialdehyde) is a key indicator to measure the degree of lipid peroxidation and oxidative stress status in the body. Hyperuricemia promotes the production of reactive oxygen species (ROS) through mechanisms such as activating NADPH oxidase, thus triggering lipid peroxidation and initiating an oxidative stress response. There is a positive correlation between the level of MDA and the concentration of uric acid. By monitoring the change of MDA level, the effect of intervention measures such as antioxidant therapy in the treatment of hyperuricemia can be evaluated. As Figure 7 shown, compared with the control group, the content of MDA in the serum of hyperuricemia (HUA) mice is significantly increased, which indicates that HUA mice have experienced lipid peroxidation reaction. Compared with the mice in the HUA group, the serum MDA content in the allopurinol group decreases, and Examples 1 to 3 can also reduce the serum MDA content, and its effect is equivalent to that of the positive control AP group. However, compared with the model group, there is no significant difference in Comparative Examples 1 to 3. This shows that the composition can improve the lipid peroxidation level of HUA mice and reduce oxidative damage. However, if any component is missing, the expected protective effect cannot be achieved.

[0105] 6 Small intestine - TCHO

[0106] HUA is a condition related to metabolic diseases, and its occurrence and development are closely related to diseases such as hyperlipidemia, hypertension and atherosclerosis. Long-term high uric acid levels may cause abnormal blood lipid levels. The total cholesterol (TCHO) level in serum was measured in the present invention, and the results are as Figure 8As shown. The results showed that compared with the blank group, the TCHO level in the serum of mice in the model group was significantly increased, indicating that dyslipidemia occurred in HUA mice; compared with the model group, allopurinol treatment could significantly reduce the TCHO level in HUA mice, indicating that allopurinol has the effect of improving the lipid level in HUA mice; after 14 days of treatment in Groups 1 to 3 of the implementation, the lipid level in HUA mice could also be significantly improved, making the blood lipid level tend to be normal, and the effect was better than that of the AP group. The same in Groups 1 to 3 of the comparison could also improve the lipid level in HUA mice, but the effect was slightly worse than that of the AP group and Groups 1 to 3 of the implementation. This indicates that the lack of any component will weaken the effect of reducing the lipid level in mice, thereby weakening its protective effect against hyperuricemia injury.

[0107] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art in this technical field, without departing from the principle of the present invention, several improvements and retouches can also be made, and these improvements and retouches should also be regarded as the protection scope of the present invention.

Claims

1. Application of N-acetylglucosamine in the preparation of health products with uric acid lowering effect.

2. A composition having the effect of lowering uric acid, characterized in that: The invention comprises the following components in parts by weight: 5 to 10 parts of N-acetylglucosamine, 2 to 5 parts of honeysuckle polysaccharide, and 1 to 3 parts of Pseudostellaria heterophylla oligosaccharide.

3. The composition according to claim 2, characterized in that The preparation method of the honeysuckle polysaccharide is as follows: extracting honeysuckle with ethanol by refluxing to obtain an extract; concentrating the extract and then precipitating with alcohol to obtain a precipitate, which is a crude honeysuckle polysaccharide; washing the crude honeysuckle polysaccharide with Sevag reagent and passing it through a cellulose chromatography column to obtain the honeysuckle polysaccharide.

4. The composition according to claim 3, characterized in that The ethanol reflux extraction method is as follows: reflux extract the honeysuckle: 95% ethanol = 1g: 5-20ml for 1-2 times, each time for 1-2h; after filtering, reflux extract the residue: 50% ethanol = 1g: 5-20ml for 2-3 times, each time for 1-2h.

5. The composition according to claim 3, characterized in that The alcohol precipitation is as follows: adding 4 to 5 times the volume of anhydrous ethanol to the concentrated extract, and standing at 4 to 6° C. for 12 to 16 hours; the washing times are 8 to 12 times.

6. The composition according to claim 2, characterized in that The preparation method of the Pseudostellariae oligosaccharide is as follows: extract Pseudostellariae with ethanol reflux to obtain an extract; concentrate the extract, extract, and precipitate with alcohol to obtain a supernatant; concentrate the supernatant and enrich it through a PA20 gel column, and according to the test results of HPLC, collect the part with a polymerization degree of 2 to 10 to obtain Pseudostellariae oligosaccharide.

7. The composition according to claim 6, characterized in that The ethanol reflux extraction method is as follows: reflux extraction is performed 2 to 3 times according to honeysuckle: 50% ethanol = 1 g: 5 to 20 ml, each time for 1 to 2 hours.

8. The composition according to claim 6, characterized in that The extraction agent used in the extraction is ethyl acetate; the volume ratio of the concentrated extract to ethyl acetate during the extraction is 1:5-20; the number of extractions is 3-4 times; the alcohol precipitation is: adding 4-5 times the volume of anhydrous ethanol to the extracted extract, standing at 4-6°C for 12-16 hours, and obtaining a supernatant.

9. The method for preparing the composition according to any one of claims 2 to 8, characterized in that: N-acetylglucosamine, honeysuckle polysaccharide, Pseudostellaria oligosaccharide and 40-60° C. water are mixed, dissolved by ultrasonication, and prepared into a solution with a concentration of 40-60 mg / mL.

10. Use of the composition according to any one of claims 2 to 8 or the composition obtained by the preparation method according to claim 9 in the preparation of a health product having the effect of lowering uric acid.

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