Composition for inhibiting fatty liver based on berry tea and preparation method thereof

Abstract

The invention discloses a composition for inhibiting fatty liver based on berry tea and a preparation method of the composition, and belongs to the technical field of functional foods. The preparation method is used for solving the technical problems that in the prior art, when berry tea and a composition thereof are applied to functional food for inhibiting fatty liver, chemical activity is unstable, and precise and continuous medical efficacy release cannot be achieved. The invention discloses a composition for inhibiting fatty liver based on berry tea. The composition comprises hydrogel loaded with active ingredients and hydrogel loaded with brown algae polyphenol. According to the hydrogel loaded with the active components, aldehyde chitosan and hyaluronic acid are used as base materials, and L-citrulline and dodecyl dimethyl betaine are used for modification, so that the dynamic crosslinking property and electrostatic adsorption capacity of the hydrogel loaded with the active components are improved. According to the hydrogel loaded with the brown algae polyphenol, the swelling property of the hydrogel is improved by constructing a double-layer network structure and a hydrogen bond network. The prepared two-component hydrogel carrier is excellent in swelling property, and the composition is high in fatty liver inhibition effect.

Description

Technical Field

[0001] The invention relates to the technical field of functional foods, and in particular to a berry tea-based composition for inhibiting fatty liver and a preparation method thereof. Background Art

[0002] Fatty liver disease is a clinical pathological syndrome characterized by abnormal accumulation of liver lipids. Fatty liver disease can be divided into non-alcoholic fatty liver disease and alcoholic fatty liver disease according to different causes; non-alcoholic fatty liver disease will deteriorate into liver fibrosis and cirrhosis, and even hepatocellular carcinoma in the later stage. Its cause is closely related to obesity, insulin resistance, intestinal flora ecological imbalance, cytokines, etc.

[0003] The scientific name of berry tea is Agave viburnum, a perennial vine plant that is mainly distributed in areas with an altitude of 200-1500m in the Yangtze River Basin of my country. The main functional ingredients of berry tea include flavonoids, polysaccharides, proteins, amino acids, etc. The results of zoological experiments show that the active ingredients in berry tea can reduce blood sugar levels, promote insulin secretion, and reduce liver glycogen content, thereby inhibiting fatty liver.

[0004] Patent application CN112220030A discloses an edible gel powder and its application. The edible gel powder includes brown algae dietary fiber and brown algae dietary fiber companion; the brown algae dietary fiber companion includes sugar, citric acid, glucono-δ-lactone and malic acid. If berry tea can be synergistically compounded with other active ingredients, the resulting compound extract can synergistically enhance the efficacy of inhibiting fatty liver. However, the stability of different natural extracts will be affected by the external environment, limiting their efficacy in the human body; in addition, how to improve the sustained-release effect of the extract to achieve the purpose of lasting inhibition of fatty liver is also a technical problem that needs to be solved urgently.

[0005] Patent application CN114698843A discloses a perilla seed oil microemulsion-hydrogel system and a preparation method thereof. The gellan gum solution is added dropwise to the chitosan-MgCl2 solution and repeatedly frozen and thawed to obtain a hydrogel; the hydrogel is mixed with the perilla seed oil microemulsion to obtain a perilla seed oil microemulsion-hydrogel system; the perilla seed oil is prepared into a hydrogel system to improve the stability and physiological activity of the perilla seed oil. If the natural active ingredients such as berry tea that inhibit fatty liver can be used as hydrogels as carriers, the stable, continuous and precise release of the medical efficacy of the natural active ingredients can be achieved. However, based on different active ingredients, the hydrogel substrates selected and prepared are also different.

[0006] In view of the technical defects in this aspect, a solution is now proposed. Summary of the invention

[0007] The purpose of the present invention is to provide a composition for inhibiting fatty liver based on berry tea and a preparation method thereof, so as to solve the technical problem that berry tea and its composition are used in functional foods for inhibiting fatty liver in the prior art, and the chemical activity is unstable and accurate and continuous release of medical efficacy cannot be achieved.

[0008] The purpose of the present invention can be achieved through the following technical solutions: A composition for inhibiting fatty liver based on berry tea, comprising, by weight, 3-5 parts of a hydrogel loaded with active ingredients and 1 part of a hydrogel loaded with brown algae polyphenols; The hydrogel loaded with active ingredients is obtained by mixing solution A and solution B; The A solution includes aldehyde-modified chitosan powder, an active ingredient solution and a PBS buffer solution, wherein the active ingredient solution includes a berry tea extract and a green tea extract solution; The B solution includes aldehyde-modified hyaluronic acid, L-citrulline, dodecyl dimethyl betaine and PBS buffer solution.

[0009] A hydrogel matrix is ​​constructed with different raw materials, and the active ingredients and brown algae polyphenols are loaded respectively, so as to obtain a two-component hydrogel, that is, the prepared berry tea-based composition for inhibiting fatty liver.

[0010] Furthermore, the method for preparing the hydrogel loaded with active ingredients comprises the following steps: A1. Crush the berry tea, pass it through a 50-mesh sieve, and dry it at 50-60° C. to constant weight to obtain berry tea powder; add ultrapure water to the berry tea powder for ultrasonic extraction, collect the extract, and concentrate the extract by rotary evaporation to obtain a berry tea extract; dissolve the green tea extract powder in deionized water to obtain a green tea extract solution; mix the berry tea extract and the green tea extract solution to obtain an active ingredient solution; The active ingredients with the highest content in berry tea are flavonoids, mainly dihydromyricetin and myricetin, which are generally 30-40%wt. After ultrasonic extraction, the yield of flavonoids in the berry tea extract is 190-200mg / g. The berry tea extract and the green tea extract solution are mixed to obtain an active ingredient solution, which is used as the active ingredient loaded on the hydrogel.

[0011] A2, aldehyde-modified chitosan powder and active ingredient solution are added to PBS buffer solution, ultrasonically dissolved to obtain solution A; aldehyde-modified hyaluronic acid, L-citrulline, and dodecyl dimethyl betaine are added to PBS buffer solution, ultrasonically dissolved to obtain solution B; A3. Mix solution A and solution B, and then let stand at room temperature until the solution becomes gel-like to obtain a hydrogel loaded with active ingredients.

[0012] PBS buffer solution is used as solvent, and aldehyde-modified chitosan powder and aldehyde-modified hyaluronic acid are used as hydrogel substrates; L-citrulline is used as a cross-linking agent, which can react with aldehyde-modified chitosan powder and aldehyde-modified hyaluronic acid to form a dynamic imine bond at room temperature. A small amount of zwitterionic dodecyl dimethyl betaine is added, which can react with the excess carboxyl, hydroxyl and amino functional groups in the aldehyde-modified chitosan and aldehyde-modified hyaluronic acid to undergo electrostatic adsorption reaction, thereby accelerating coagulation and obtaining a hydrogel loaded with active ingredients.

[0013] Furthermore, in step A1, the ratio of berry tea powder to deionized water is 5-10 g:50 mL, the ratio of green tea extract powder to deionized water is 3-5 g:10 mL, and the ratio of berry tea extract to green tea extract solution is 10-20 mL:10 mL.

[0014] Furthermore, in step A2, the concentration of PBS buffer solution is 1×PBS, pH=7-7.4; the dosage ratio of aldehyded chitosan powder, active ingredient solution and PBS buffer solution is 20-30 mg:10-20 mg:50 mL; the dosage ratio of aldehyded hyaluronic acid, L-citrulline, dodecyl dimethyl betaine and PBS buffer solution is 15-20 mg:2-5 mg:2-5 mg:50 mL.

[0015] The concentration of PBS buffer solution was set to 0.01 mol / L and the pH value was set to 7-7.4 to simulate the in vivo environment.

[0016] Furthermore, the method for preparing the hydrogel loaded with brown algae polyphenols comprises the following steps: B1. Glucose, coumaric acid and deionized water with a pH value of 3-4 are added to a reactor and mixed evenly; in an inert gas atmosphere, the reactor is heated to 150-200° C. and kept at this temperature for 3-5 hours, and the water generated during the reaction is removed in time; the reactor is then cooled and allowed to stand at room temperature to obtain a copolymer; Glucose and coumaric acid are used as monomers, and esterification reaction occurs to obtain a copolymer.

[0017] B2. Add brown algae polyphenols and linoleic acid to the copolymer, then heat the reactor to 80-90° C., and react at this temperature for 10-12 hours to obtain a brown algae polyphenol-loaded hydrogel.

[0018] The active ingredient brown algae polyphenol is added to the copolymer, and under high temperature conditions, the copolymer and linoleic acid undergo addition reaction to obtain a brown algae polyphenol-loaded hydrogel.

[0019] Furthermore, in step B1, the usage ratio of glucose, coumaric acid and deionized water is 80-100 g:18-36 g; 200 mL; in step B2, the usage ratio of brown algae polyphenols and linoleic acid is 10-20 g:5-10 g.

[0020] As another aspect of the present invention, a method for preparing a composition for inhibiting fatty liver based on berry tea comprises the following steps: The hydrogel loaded with active ingredients and the hydrogel loaded with brown algae polyphenols are compounded to obtain a berry tea-based composition for inhibiting fatty liver.

[0021] The two-component hydrogel is compounded to obtain a berry tea-based composition for inhibiting fatty liver.

[0022] As another aspect of the present invention, a composition for inhibiting fatty liver based on berry tea is used in the preparation of a functional food having the effect of inhibiting fatty liver.

[0023] The present invention has the following beneficial effects: Hydrogels are closer to living tissues than any synthetic biomaterials, and their properties are similar to those of extracellular matrix, and they can be used as storage compartments and delivery tools for drugs. Hydrogels are used to load compositions that inhibit fatty liver, and active ingredients can be delivered in a targeted manner, thereby improving the delivery efficiency of drug components. The present invention compounded a hydrogel loaded with active ingredients and a hydrogel loaded with brown algae polyphenols to obtain a two-component hydrogel, which can be added to functional foods as a composition that inhibits fatty liver. Brown algae polyphenols can be utilized by intestinal flora to enrich beneficial intestinal flora and reduce the abundance of harmful flora associated with metabolic diseases such as diabetes. However, as a polyhydroxy compound, brown algae polyphenols are prone to chemical reactions such as oxidation and degradation, and using hydrogels to load brown algae polyphenols can improve their own stability. When preparing a hydrogel loaded with brown algae polyphenols, glucose and coumaric acid are subjected to an esterification reaction to obtain a first layer of network structure; linoleic acid is then added to form a second layer of network structure through an addition reaction, thereby obtaining a layer-by-layer self-assembled hydrogel structure. Since the active ingredient brown algae polyphenol loaded in the hydrogel is a polyphenol structure, it can form a hydrogen bond network with glucose, thereby improving the mechanical properties and swelling properties of the prepared hydrogel. The high swelling properties of the hydrogel are used to achieve controlled release of the active substance, thereby improving the utilization rate of the active ingredient. 3. Berry tea extract has the effect of protecting the liver and can improve diet-induced non-alcoholic fatty hepatitis; by assisting in lowering blood sugar and blood lipids, berry tea extract can help diabetics achieve blood sugar control and improve renal function. Green tea extract contains many biologically active substances, such as free amino acids, tea polyphenols and flavonoids; berry tea extract and green tea extract are used as active substances for inhibiting fatty liver and then loaded into the hydrogel, which can achieve the effect of synergistically inhibiting fatty liver and enhancing the therapeutic effect. The substrate for loading the active ingredient solution of the present invention is chitosan and hyaluronic acid. Chitosan itself has good swelling properties and biocompatibility, but chitosan has poor water solubility. Chitosan can improve its own water solubility through aldehyde reaction. Hyaluronic acid is surrounded by a sphere of water molecules connected by hydrogen bonds and is highly hydrophilic. After being aldehyde-formylated, chitosan and hyaluronic acid can react with L-citrulline to form a dynamic imine bond, thereby rapidly undergoing a self-assembly cross-linking reaction at room temperature to form a hydrogel. In addition, the synthesized dynamic imine bond is used to provide the hydrogel with self-healing ability, thereby improving its own stability. When preparing a hydrogel loaded with an active ingredient solution, a small amount of dodecyl dimethyl betaine is added in the present invention. Dodecyl dimethyl betaine, as a zwitterion, can undergo an electrostatic adsorption reaction with excess carboxyl, hydroxyl and amino functional groups in the aldehyde-formylated chitosan and aldehyde-formylated hyaluronic acid, thereby accelerating coagulation; since the action between anions and cations is reversible, the prepared hydrogel has a unique ability to self-repair and respond to external stimuli. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0025] Figure 1 The present invention is a process flow chart for preparing a berry tea-based composition for inhibiting fatty liver. DETAILED DESCRIPTION

[0026] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0027] The CAS number of chitosan used in Example 1 of the present invention is 9012-76-4, which was purchased from Zhengzhou Dongyi Chemical Co., Ltd. with a product number of 612636-631; the CAS number of hyaluronic acid used in Example 2 is 9004-61-9; the green tea extract powder used in Examples 3-5 of the present invention was purchased from Xi'an Beijinuo Biotechnology Co., Ltd. with a product number of BJN0893.

[0028] Example 1 This embodiment provides a method for preparing aldehyde-modified chitosan powder for loading active ingredients, comprising the following steps: 1. Add 120 mL of deionized water to a 250 mL beaker, then add 0.1 mol / L HCl solution to the deionized water to adjust the pH value of the deionized water to 4, then add 3 g of chitosan to the beaker and continue stirring until the chitosan is completely dissolved in the deionized water to obtain a chitosan aqueous solution.

[0029] 2. Add 0.5 mL of 0.5 mol / L sodium periodate solution to the chitosan aqueous solution and mix well to obtain a mixed solution. Transfer the beaker to a water bath, and then transfer the water bath to a fume hood for light protection, accompanied by magnetic stirring at 200 r / min for 3 hours to obtain a reaction solution; add 2 mL of ethylene glycol to the reaction solution to terminate the reaction and obtain the product.

[0030] 3. The product is dialyzed for 24 hours using a dialysis bag with a molecular weight cutoff of 10,000 Da to obtain a dialyzed solution; the dialyzed solution is concentrated by rotary evaporation to obtain a concentrated solution; the concentrated solution is freeze-dried to obtain aldehyde-modified chitosan powder.

[0031] Example 2 This embodiment provides a method for preparing aldehyde-modified hyaluronic acid for loading active ingredients, comprising the following steps: Add 150mL of deionized water to a 250mL beaker, then drop 0.1mol / L HCl solution into the deionized water, adjust the pH value of the deionized water to 6, then add 5g of hyaluronic acid to the beaker, and continue stirring until the hyaluronic acid is completely dissolved in the deionized water to obtain a hyaluronic acid aqueous solution. Add 2mL of 0.5mol / L sodium periodate solution to the hyaluronic acid aqueous solution, mix well, and obtain a mixed solution. Transfer the beaker to a water bath, and then transfer the water bath to a fume hood for light protection, accompanied by magnetic stirring at 200r / min for 3h to obtain a reaction solution; add 2mL of ethylene glycol to the reaction solution to terminate the reaction and obtain a product. The product is dialyzed for 24h using a dialysis bag with a molecular weight cutoff of 6000Da to obtain a dialyzed solution; the dialyzed solution is concentrated by rotary evaporation to obtain a concentrated solution; the concentrated solution is freeze-dried to obtain aldehyde-modified hyaluronic acid powder. Example 3 This embodiment provides a method for preparing a hydrogel loaded with active ingredients for a composition for inhibiting fatty liver based on berry tea, comprising the following steps: A1. Weigh 5g of berry tea, crush it with a grinder, and then pass it through a 50-mesh sieve to obtain powder; the powder is dried to constant weight at 50°C to obtain berry tea powder. Mix 5g of berry tea powder with 50mL of deionized water, ultrasonically extract at 80°C, wherein the ultrasonic power is 350W and the ultrasonic time is 40min, and collect the extract; the extract is concentrated by rotary evaporation to obtain 20mL of berry tea extract. Mix 3g of green tea extract and 10mL of deionized water, and shake to mix well to obtain a green tea extract solution; mix 10mL of berry tea extract and 10mL of green tea extract solution to obtain an active ingredient solution.

[0032] A2. Add 20 mg of the aldehyde-modified chitosan powder prepared in Example 1 and 10 mg of the active ingredient solution to 50 mL of 1×PBS, pH=7 PBS buffer solution, and dissolve them by ultrasound to obtain solution A; add 15 mg of the aldehyde-modified hyaluronic acid prepared in Example 2, 2 mg of L-citrulline and 5 mg of dodecyl dimethyl betaine to 50 mL of 1×PBS, pH=7.2 PBS buffer solution, and dissolve them by ultrasound to obtain solution B.

[0033] A3. Add 20 mL of solution A and 10 mL of solution B into a 50 mL centrifuge tube, mix well, and let stand at room temperature until the solution in the centrifuge tube is in a gel state to obtain a hydrogel loaded with active ingredients.

[0034] Example 4 This embodiment provides a method for preparing a hydrogel loaded with active ingredients for a composition for inhibiting fatty liver based on berry tea, comprising the following steps: A1. Weigh 6g of berry tea, crush it with a grinder, and then pass it through a 50-mesh sieve to obtain powder; the powder is dried at 57°C to constant weight to obtain berry tea powder. Mix 6g of berry tea powder with 50mL of deionized water, ultrasonically extract at 88°C, wherein the ultrasonic power is 450W and the ultrasonic time is 50min, and collect the extract; the extract is concentrated by rotary evaporation to obtain 20mL of berry tea extract. Mix 4g of green tea extract powder and 10mL of deionized water, and oscillate to obtain a green tea extract solution; mix 15mL of berry tea extract and 10mL of green tea extract solution to obtain an active ingredient solution.

[0035] A2. Add 25 mg of the aldehyde-modified chitosan powder prepared in Example 1 and 15 mg of the active ingredient solution to 50 mL of 1×PBS, pH=7.4 PBS buffer solution, and dissolve them by ultrasound to obtain solution A; add 18 mg of the aldehyde-modified hyaluronic acid prepared in Example 2, 3 mg of L-citrulline and 3 mg of dodecyl dimethyl betaine to 50 mL of 1×PBS, pH=7 PBS buffer solution, and dissolve them by ultrasound to obtain solution B.

[0036] A3. Add 18 mL of solution A and 10 mL of solution B into a 50 mL centrifuge tube, mix well, and let stand at room temperature until the solution in the centrifuge tube is in a gel state to obtain a hydrogel loaded with active ingredients.

[0037] Example 5 This embodiment provides a method for preparing a hydrogel loaded with active ingredients for a composition for inhibiting fatty liver based on berry tea, comprising the following steps: A1. Weigh 10g of berry tea, crush it with a grinder, and then pass it through a 50-mesh sieve to obtain powder; the powder is dried at 60°C to constant weight to obtain berry tea powder. Mix 10g of berry tea powder with 50mL of deionized water, perform ultrasonic extraction at 90°C, wherein the ultrasonic power is 500W and the ultrasonic time is 60min, and collect the extract; the extract is concentrated by rotary evaporation to obtain 20mL of berry tea extract. Mix 5g of green tea extract powder and 10mL of deionized water, shake and mix to obtain a polypeptide solution; mix 20mL of berry tea extract and 10mL of green tea extract solution to obtain an active ingredient solution.

[0038] A2. Add 30 mg of the aldehyde-modified chitosan powder prepared in Example 1 and 20 mg of the active ingredient solution to 50 mL of 1×PBS, pH=7.2 PBS buffer solution, and dissolve them by ultrasound to obtain solution A; add 20 mg of the aldehyde-modified hyaluronic acid prepared in Example 2, 5 mg of L-citrulline and 5 mg of dodecyl dimethyl betaine to 50 mL of 1×PBS, pH=7.2 PBS buffer solution, and dissolve them by ultrasound to obtain solution B.

[0039] A3. Add 15 mL of solution A and 10 mL of solution B into a 50 mL centrifuge tube, mix well, and let stand at room temperature until the solution in the centrifuge tube is in a gel state to obtain a hydrogel loaded with active ingredients.

[0040] Example 6 This embodiment provides a method for preparing a hydrogel loaded with brown algae polyphenols for a composition for inhibiting fatty liver based on berry tea, comprising the following steps: B1. Measure 200 mL of deionized water, add 0.1 mol / L HCl solution to the deionized water, and adjust the pH value of the deionized water to 3. Add 200 mL of deionized water, 50 g of glucose and 18 g of coumaric acid to a 500 mL enamel reactor in sequence, mix and stir evenly; connect the enamel reactor to an inert gas inlet pipe, and introduce nitrogen into the enamel reactor to replace the air in the reactor; connect the reactor to a dropping funnel, insert an oil-water separator and a condenser, heat it to 150°C, keep it warm for 3 hours at this temperature, and remove the water produced in the esterification reaction in time during the reaction. Then let the reactor stand and cool to room temperature to obtain a copolymer.

[0041] B2. Add 10 g of brown algae polyphenols and 5 g of linoleic acid into an enameled reactor, then raise the temperature of the reactor to 80° C. and react at this temperature for 10 h to obtain a hydrogel loaded with brown algae polyphenols.

[0042] B3. According to parts by weight, 3 parts of the hydrogel loaded with active ingredients prepared in Example 5 and 1 part of the hydrogel loaded with brown algae polyphenols were compounded to obtain a berry tea-based composition for inhibiting fatty liver.

[0043] Example 7 This embodiment provides a method for preparing a hydrogel loaded with brown algae polyphenols for a composition for inhibiting fatty liver based on berry tea, comprising the following steps: B1. Measure 200 mL of deionized water, add 0.1 mol / L HCl solution to the deionized water, and adjust the pH value of the deionized water to 4. Add 200 mL of deionized water, 55 g of glucose and 28 g of coumaric acid to a 500 mL enamel reactor in sequence, mix and stir evenly; connect the enamel reactor to an inert gas inlet pipe, and introduce nitrogen into the enamel reactor to replace the air in the reactor; connect the reactor to a dropping funnel, insert an oil-water separator and a condenser, heat it to 175°C, keep it warm for 3.5 hours at this temperature, and remove the water produced in the reaction in time during the reaction. Then let the reactor stand and cool to room temperature to obtain a copolymer.

[0044] B2. Add 16 g of brown algae polyphenols, 6 g of linoleic acid and 0.02 g of initiator benzoyl peroxide into an enamel reactor, then raise the temperature of the reactor to 85° C. and react at this temperature for 11 hours to obtain a hydrogel loaded with brown algae polyphenols.

[0045] B3. According to parts by weight, 4 parts of the hydrogel loaded with active ingredients prepared in Example 5 and 1 part of the hydrogel loaded with brown algae polyphenols were compounded to obtain a berry tea-based composition for inhibiting fatty liver.

[0046] Example 8 This embodiment provides a method for preparing a hydrogel loaded with brown algae polyphenols for a composition for inhibiting fatty liver based on berry tea, comprising the following steps: B1. Measure 200 mL of deionized water, add 0.1 mol / L HCl solution to the deionized water, and adjust the pH value of the deionized water to 4. Add 200 mL of deionized water, 60 g of glucose and 36 g of coumaric acid to a 500 mL enamel reactor in sequence, mix and stir evenly; connect the enamel reactor to an inert gas inlet pipe, and introduce nitrogen into the enamel reactor to replace the air in the reactor; connect the reactor to a dropping funnel, insert an oil-water separator and a condenser, heat it to 200°C, keep it warm for 5 hours at this temperature, and remove the water produced in the reaction in time during the reaction. Then let the reactor stand and cool to room temperature to obtain a copolymer.

[0047] B2. Add 20 g of brown algae polyphenols, 10 g of linoleic acid and 0.03 g of initiator benzoyl peroxide into an enamel reactor, then raise the temperature of the reactor to 90° C. and react at this temperature for 12 h to obtain a hydrogel loaded with brown algae polyphenols.

[0048] B3. According to parts by weight, 5 parts of the hydrogel loaded with active ingredients prepared in Example 5 and 1 part of the hydrogel loaded with brown algae polyphenols were compounded to obtain a berry tea-based composition for inhibiting fatty liver.

[0049] Comparative Example 1 The difference between this comparative example and Example 8 is that, when preparing the hydrogel loaded with active ingredients, the aldehyde-modified chitosan powder is replaced by the chitosan powder of the same mass, and the aldehyde-modified hyaluronic acid is replaced by the hyaluronic acid of the same mass.

[0050] Comparative Example 2 The difference between this comparative example and Example 8 is that when preparing the hydrogel loaded with active ingredients, sodium dodecylbenzenesulfonate of the same mass is used to replace dodecyl dimethyl betaine.

[0051] Comparative Example 3 The difference between this comparative example and Example 8 is that when preparing the hydrogel loaded with brown algae polyphenols, linoleic acid is not added to the copolymer.

[0052] Comparative Example 4 The difference between this comparative example and Example 8 is that when preparing the hydrogel loaded with active ingredients, the active ingredient solution is replaced by an equal mass of berry tea extract.

[0053] Performance Test: The composition for inhibiting fatty liver based on berry tea prepared in Examples 6-8 and Comparative Examples 1-3 was freeze-dried and weighed; then immersed in a PBS buffer solution with a pH of 7.0, and the concentration of the PBS buffer solution was 1×PBS; after reaching swelling equilibrium, it was taken out and weighed again. The calculation formula of the equilibrium swelling ratio of the hydrogel is as follows: ESR (equilibrium swelling ratio) = (W S -W d / W d )×100% Among them, W S W is the mass of the composition for inhibiting fatty liver based on berry tea after swelling equilibrium; d This is the mass of the berry tea-based composition for inhibiting fatty liver before swelling.

[0054] Table 1- Sample test data

[0055] Data analysis: A comparative analysis of the data in Table 1 was performed using a PBS buffer solution with a pH of 7.0 and a concentration of 1×PBS to simulate the human body environment. The compositions for inhibiting fatty liver based on berry tea prepared in Examples 6-8 of the present invention all had excellent swelling properties, as shown by a higher value of the equilibrium swelling ratio. Aldehyde-formylated chitosan and aldehyde-formylated hyaluronic acid can react with L-citrulline to undergo a Schiff base reaction, and the dynamic imine bond is used to increase the crosslinking density of the two substrate components. However, in Comparative Example 1, the aldehyde-formylated substrate raw material is replaced by an unaldehyde-formylated substrate, thereby reducing the crosslinking density of the two substrates, thereby reducing the crosslinking density of the two substrates themselves.

[0056] Zwitterions can react with the excess functional groups in the substrate hyaluronic acid and chitosan to undergo electrostatic adsorption reaction, thereby improving the coagulation performance. In Comparative Example 2, sodium dodecylbenzene sulfonate of equal mass was used to replace dodecyl dimethyl betaine. Since sodium dodecylbenzene sulfonate does not have anions or cations, the electrostatic adsorption reaction was reduced, the cross-linking density thereof was reduced, and the swelling performance thereof was reduced. In Comparative Example 3, linoleic acid was not added to the prepared copolymer, and the second layer of network structure was not formed, thereby reducing the swelling performance of the hydrogel loaded with brown algae polyphenols, which was manifested as a decrease in the value of the self-equilibrium swelling ratio.

[0057] Clean-grade, male SD rats were selected, and the mass of each rat was 200g. The above rats were divided into low-dose group, medium-dose group, high-dose group, control group 1 and control group 2, with three rats in each group. The mice in the low-dose group were gavaged with a composition for inhibiting fatty liver based on berry tea prepared in Example 8 at 2.5 mg / kg; the mice in the medium-dose group were gavaged with a composition for inhibiting fatty liver based on berry tea prepared in Example 8 at 5 mg / kg; the mice in the high-dose group were gavaged with a composition for inhibiting fatty liver based on berry tea prepared in Example 8 at 10 mg / kg; the control group 1 was gavaged with 0.5%wt normal saline; the control group 2 was gavaged with a composition for inhibiting fatty liver based on berry tea prepared in Comparative Example 4. All treatments were gavaged once a day for one week, and then the levels of ALT, AST and T-CHO in the mouse serum were measured. Table 2-Sample test data

[0058] Data analysis: A comparative analysis of the data in Table 2 shows that a composition for inhibiting fatty liver based on berry tea prepared in Example 8 of the present invention can regulate liver tissue metabolic disorders and liver inflammation in mice, and has a liver-protecting effect, which is manifested in that the ALT, AST and T-CHO content in the serum of mice in the medium and high-dose groups are lower. Reduced levels of ALT, AST and T-CHO in serum can effectively alleviate non-alcoholic fatty liver damage and inhibit the formation of fatty liver. However, in Control Group 2, the synergistic active ingredient green tea extract was removed, thereby reducing its own pharmacological efficacy, which is manifested in that the ALT, AST and T-CHO content in the serum of mice is higher than that in the medium and high-dose groups.

[0059] The above contents are merely examples and explanations of the structure of the present invention. The technicians in this technical field may make various modifications or additions to the specific embodiments described or replace them in a similar manner. As long as they do not deviate from the structure of the invention or exceed the scope defined by the claims, they should all fall within the protection scope of the present invention.

[0060] In the description of this specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0061] The preferred embodiments of the present invention disclosed above are only used to help explain the present invention. The preferred embodiments do not describe all the details in detail, nor do they limit the invention to only specific implementation methods. Obviously, many modifications and changes can be made according to the content of this specification. This specification selects and specifically describes these embodiments in order to better explain the principles and practical applications of the present invention, so that those skilled in the art can understand and use the present invention well. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. A composition for inhibiting fatty liver based on berry tea, characterized in that: According to weight, it includes 3-5 parts of hydrogel loaded with active ingredients and 1 part of hydrogel loaded with brown algae polyphenols; The hydrogel loaded with active ingredients is obtained by mixing solution A and solution B; The A solution comprises aldehyde-modified chitosan powder, an active ingredient solution and a PBS buffer solution, wherein the active ingredient solution comprises a berry tea extract and a green tea extract solution; The B solution includes aldehyde-modified hyaluronic acid, L-citrulline, dodecyl dimethyl betaine and PBS buffer solution.

2. A composition for inhibiting fatty liver based on berry tea according to claim 1, characterized in that: The method for preparing the hydrogel loaded with active ingredients comprises the following steps: A1. Crush the berry tea, sieve it, and dry it at 50-60° C. to constant weight to obtain berry tea powder; ultrasonically extract the berry tea powder with deionized water, collect the extract, and concentrate the extract by rotary evaporation to obtain a berry tea extract; dissolve the green tea extract with deionized water to obtain a green tea extract solution; mix the berry tea extract and the green tea extract solution to obtain an active ingredient solution; A2, aldehyde-modified chitosan powder and active ingredient solution are added to PBS buffer solution and ultrasonically dissolved to obtain solution A; aldehyde-modified hyaluronic acid, L-citrulline and dodecyl dimethyl betaine are added to PBS buffer solution and ultrasonically dissolved to obtain solution B; A3. Mix solution A and solution B, and then let stand at room temperature until the solution becomes gel-like to obtain a hydrogel loaded with active ingredients.

3. The composition for inhibiting fatty liver based on berry tea according to claim 2, characterized in that: In step A1, the dosage ratio of berry tea powder to deionized water is 5-10 g:50 mL, the dosage ratio of green tea extract powder to deionized water is 3-5 g:10 mL, and the dosage ratio of berry tea extract to green tea extract solution is 10-20 mL:10 mL.

4. The composition for inhibiting fatty liver based on berry tea according to claim 2, characterized in that: In step A2, the concentration of PBS buffer solution is 1×PBS, pH=7-7.4; the dosage ratio of aldehyded chitosan powder, active ingredient solution and PBS buffer solution is 20-30 mg:10-20 mg:50 mL; the dosage ratio of aldehyded hyaluronic acid, L-citrulline, dodecyl dimethyl betaine and PBS buffer solution is 15-20 mg:2-5 mg:2-5 mg:50 mL.

5. The composition for inhibiting fatty liver based on berry tea according to claim 1, characterized in that: The preparation method of the hydrogel loaded with brown algae polyphenols, The following steps are involved: B1, adding glucose, coumaric acid and deionized water into a reactor, mixing them well, placing the reactor under an inert gas atmosphere and heating it to 150-200° C., keeping the temperature at this temperature for 3-5 hours, then cooling the reactor to room temperature, collecting the lower precipitate, and obtaining a copolymer; B2. Add brown algae polyphenols and linoleic acid to the copolymer, then heat the reactor to 80-90° C., and react at this temperature for 10-12 hours to obtain a brown algae polyphenol-loaded hydrogel.

6. The composition for inhibiting fatty liver based on berry tea according to claim 5, characterized in that: In step B1, the usage ratio of glucose, coumaric acid and deionized water is 50-60 g:16-32 g:200 mL; in step B2, the usage ratio of brown algae polyphenols and linoleic acid is 10-20 g:5-10 g.

7. A method for preparing a composition for inhibiting fatty liver based on berry tea as claimed in any one of claims 1 to 6, characterized in that: The following steps are involved: A berry tea-based composition for inhibiting fatty liver is obtained by compounding a hydrogel loaded with active ingredients and a hydrogel loaded with brown algae polyphenols.

8. An application of a composition for inhibiting fatty liver based on berry tea, characterized in that: Use of a berry tea-based fatty liver inhibiting composition according to any one of claims 1 to 6 in the preparation of a functional food having the effect of inhibiting fatty liver.

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