Thaflavin composition for treating alcoholic liver injury as well as preparation method and application of theaflavin composition

Abstract

The invention provides a theaflavin composition for treating alcoholic liver injury as well as a preparation method and application thereof, and relates to the technical field of medicines. The theaflavin composition comprises theaflavin, a radix puerariae extract, a hovenia dulcis thunb extract, a coreopsis tinctoria extract, a kelp extract, a licorice extract and an antioxidant. Wherein the theaflavin, the senecio cineraria extract and the antioxidant can reduce oxidative stress, the radix puerariae extract, the liquorice extract and the hovenia dulcis thunb extract have an anti-inflammatory effect, the radix puerariae extract and the hovenia dulcis thunb extract promote alcohol metabolism, and the kelp extract and the liquorice extract have an effect of promoting liver cell regeneration. All the components of the theaflavin composition can effectively relieve alcoholic liver injury through multi-target and multi-path synergistic effects, and the theaflavin composition has remarkable liver protection and anti-inflammatory effects.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a theaflavin composition for treating alcoholic liver damage, and a preparation method and application thereof. Background Art

[0002] Alcoholic liver damage is a liver disease caused by long-term excessive drinking, posing a serious threat to the health of numerous people worldwide. Long-term alcohol consumption places an excessive metabolic burden on the liver, triggering a series of pathological changes. Initially, alcoholic fatty liver disease may develop, manifesting as fat accumulation within liver cells. Continued drinking can further develop into alcoholic hepatitis, characterized by significant liver inflammation and increased liver cell damage. If the condition remains uncontrolled, it can eventually worsen into alcoholic cirrhosis and even liver cancer. According to relevant medical research statistics, the incidence of alcoholic liver damage among alcoholics has been increasing year by year, becoming a major cause of liver disease and placing a heavy burden on social medical resources. Currently, the treatment options for alcoholic liver damage are relatively limited. Abstinence from alcohol is undoubtedly the most important measure for treating alcoholic liver disease, but for some patients with more severe liver damage, simply abstaining from alcohol is often difficult to completely reverse the pathological changes in the liver. In terms of drug treatment, although there are some liver-protecting drugs, such as polyene phosphatidylcholine capsules, which can accelerate the repair and regeneration of damaged liver cell membranes to a certain extent and prevent the disease from worsening, the overall efficacy is still not ideal. In addition, drugs such as metadoxine, corticosteroids, and S-adenosylmethionine have problems such as insignificant effects, high costs, or large side effects, which prevents them from being widely used. Although liver transplantation can temporarily alleviate the problem during the decompensated stage of the liver, it is not only expensive and risky, but also has a very limited source of liver donors. In recent years, with the continued deepening of research on natural products, it has been discovered that some functional components in tea have potential hepatoprotective effects. However, current research on tea's functional components, both domestically and internationally, primarily focuses on tea, crude tea extracts, or single catechin components, with a lack of research on the effects of other tea components on alcoholic liver injury. Furthermore, single-component medications have limited functionality and cannot comprehensively treat alcoholic liver injury. Therefore, the development of a theaflavin composition for the treatment of alcoholic liver injury, as well as its preparation and application, is of great practical significance and holds broad market potential. A composition containing theaflavins as its primary ingredient, with significant efficacy, safety, and reliability, for the treatment of alcoholic liver injury, possesses significant practical significance and broad market prospects. Summary of the Invention

[0003] The purpose of the present invention is to provide a theaflavin composition for treating alcoholic liver damage, and its preparation method and application. The theaflavin composition provided by the present invention has significant therapeutic effects in treating alcoholic liver damage.

[0004] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions: The present invention provides a theaflavins composition for treating alcoholic liver damage, comprising the following raw materials in parts by weight: 20-30 parts of theaflavins, 20-30 parts of kudzu root extract, 15-25 parts of hovenia dulcis fruit extract, 10-20 parts of snow chrysanthemum extract, 5-10 parts of kelp extract, 3-8 parts of liquorice extract and 0.01-0.03 parts of antioxidant.

[0005] Preferably, the preparation method of the kudzu root extract comprises: crushing the kudzu root, mixing it with water, inoculating a composite bacterial agent for liquid anaerobic fermentation, sterilizing, filtering, concentrating the filtrate, and drying to obtain the kudzu root extract.

[0006] More preferably, the composite bacterial agent consists of Lactobacillus hilgris GZ2 and Bifidobacterium bifidum CICC 6169.

[0007] More preferably, the temperature of the liquid fermentation is 35-40° C., the pH value is 5-6, and the time is 58-72 hours.

[0008] Preferably, the Hovenia dulcis fruit extract, Snow chrysanthemum extract and Licorice root extract are obtained by extraction with ethanol solution.

[0009] Preferably, the kelp extract is extracted by a high-voltage pulse electric field method.

[0010] Preferably, the antioxidant is vitamin C or vitamin E.

[0011] The present invention also provides a method for preparing the theaflavins composition, comprising: mixing antioxidants, theaflavins, kudzu root extract, hovenia dulcis fruit extract, snow chrysanthemum extract, kelp extract and liquorice extract to obtain the theaflavins composition.

[0012] The present invention also provides the use of the theaflavin composition in preparing a drug for treating liver damage.

[0013] Preferably, the liver injury is alcoholic liver injury.

[0014] Compared with the prior art, the present invention has the following beneficial effects: The present invention provides a theaflavins composition for treating alcoholic liver injury, comprising theaflavins, kudzu root extract, Hovenia dulcis fruit extract, snow chrysanthemum extract, kelp extract, licorice extract, and an antioxidant. The theaflavins, snow chrysanthemum extract, and antioxidant reduce oxidative stress; the kudzu root extract, licorice extract, and Hovenia dulcis fruit extract exhibit anti-inflammatory effects; the kudzu root extract and Hovenia dulcis fruit extract promote alcohol metabolism; and the kelp extract and licorice extract promote hepatocyte regeneration. The components of the theaflavins composition work synergistically across multiple targets and pathways, effectively alleviating alcoholic liver injury and exhibiting significant hepatoprotective, anti-inflammatory, and antioxidant effects. DETAILED DESCRIPTION

[0015] The present invention provides a theaflavins composition for treating alcoholic liver damage, comprising the following raw materials in parts by weight: 20-30 parts of theaflavins, 20-30 parts of kudzu root extract, 15-25 parts of hovenia dulcis fruit extract, 10-20 parts of snow chrysanthemum extract, 5-10 parts of kelp extract, 3-8 parts of liquorice extract and 0.01-0.03 parts of antioxidant.

[0016] The method for preparing the kudzu root extract of the present invention preferably comprises: crushing the kudzu root, mixing it with water, inoculating a composite bacterial agent for liquid anaerobic fermentation, sterilizing, filtering, concentrating the filtrate, and drying to obtain the kudzu root extract. The composite bacterial agent is preferably composed of Lactobacillus hirsutigensis GZ2 and Bifidobacterium bifidum CICC 6169, the bacterial count of Lactobacillus hirsutigensis GZ2 in the composite bacterial agent is preferably 500-1 billion cells / g, more preferably 800 million cells / g, and the bacterial count of Bifidobacterium bifidum CICC 6169 is preferably 200-500 million cells / g, more preferably 300 million cells / g; the temperature of the liquid fermentation is preferably 35-40°C, more preferably 37°C, the pH value is preferably 5-6, more preferably 5.5, and the fermentation time is preferably 58-72 hours, more preferably 60 hours. The present invention uses specific bacterial species Lactobacillus hilgeni GZ2 and Bifidobacterium bifidum CICC 6169 to ferment kudzu root. On the one hand, the microorganisms can use the kudzu root as an energy substance for decomposition. On the other hand, through the fermentation of the microbial agent, the components in the kudzu root can be converted into new metabolites, so that the obtained kudzu root extract has a better therapeutic effect on alcohol-fat damage.

[0017] The Hovenia dulcis fruit extract, snow chrysanthemum extract and liquorice extract of the present invention are preferably obtained by extraction with ethanol solution.

[0018] The preparation method of the Hovenia dulcis fruit extract of the present invention preferably comprises: pulverizing the Hovenia dulcis fruit, adding a 65%-75% ethanol solution by volume at a solid-liquid ratio of 1:8-12, and ultrasonically extracting the extract at a temperature of 48-52°C, a power of 250-350W, and a frequency of 23-28kHz for 25-35 minutes. More preferably, the method comprises: pulverizing the Hovenia dulcis fruit, adding a 70% ethanol solution by volume at a solid-liquid ratio of 1:10, and ultrasonically extracting the extract at a temperature of 50°C, a power of 300W, and a frequency of 25kHz for 30 minutes. The Hovenia dulcis fruit extract of the present invention has the effects of accelerating alcohol metabolism, alleviating symptoms of intoxication (such as headache and nausea), and reducing the risk of liver cell damage; providing antioxidant properties, alleviating oxidative stress, and preventing alcohol-induced liver cell necrosis; and providing anti-inflammatory properties, inhibiting liver damage.

[0019] The preparation method of the snow chrysanthemum extract of the present invention preferably comprises: crushing the snow chrysanthemum, adding a 55%-65% ethanol solution by volume at a material-liquid ratio of 10-15, and ultrasonically extracting at a temperature of 43-48°C, a power of 200-250W, and a frequency of 25-30kHz for 30-40 minutes. More preferably, the method comprises: crushing the snow chrysanthemum, adding a 60% ethanol solution by volume at a material-liquid ratio of 1:12, and ultrasonically extracting at a temperature of 45°C, a power of 230W, and a frequency of 28kHz for 35 minutes. The snow chrysanthemum extract of the present invention has antioxidant activity, reduces liver tissue inflammation, and improves liver function.

[0020] The preparation method of the licorice extract of the present invention preferably comprises: crushing the licorice, adding a 48%-53% ethanol solution by volume at a solid-liquid ratio of 1:15-20, and ultrasonically extracting at a temperature of 38-45°C, a power of 250-300W, and a frequency of 30-35kHz for 40-50 minutes; more preferably, it comprises: crushing the licorice, adding a 50% ethanol solution by volume at a solid-liquid ratio of 1:18, and ultrasonically extracting at a temperature of 42°C, a power of 280W, and a frequency of 32kHz for 45 minutes. The licorice extract of the present invention can enhance the liver's detoxification function, promote alcohol metabolism, have a strong anti-inflammatory effect, protect liver cells, and improve alcohol-induced immune suppression, enhancing the liver's defense against pathogens.

[0021] The preparation method of the kelp extract of the present invention preferably includes: mixing kelp powder and water at a solid-liquid ratio of 1:20-30 g / mL, extracting with a high-voltage pulse electric field at an electric field strength of 20-30 kV / cm, a pulse width of 5-10 μs, and a temperature of 70-80°C for 30-60 seconds, filtering, concentrating the filtrate to 1 / 2-1 / 4 of the volume of the supernatant, adding 8-12 times the volume of an ethanol solution with a volume fraction of 90%-97%, precipitating for 10-14 hours, centrifuging, and drying the precipitate to obtain the kelp extract; more preferably includes: mixing kelp powder and water at a solid-liquid ratio of 1:25 g / mL, extracting with a high-voltage pulse electric field at an electric field strength of 25 kV / cm, a pulse width of 8 μs, and a temperature of 75°C for 45 seconds, filtering, concentrating the filtrate to 1 / 3 of the volume of the supernatant, adding 10 times the volume of an ethanol solution with a volume fraction of 95%, precipitating for 12 hours, centrifuging, and drying the precipitate to obtain the kelp extract. The kelp extract of the present invention has antioxidant activity, can repair intestinal damage, regulate intestinal flora, and improve liver inflammation damage caused by alcohol.

[0022] The antioxidant of the present invention is preferably vitamin C or vitamin E, more preferably vitamin C. On the one hand, the antioxidant can protect other active ingredients from oxidation when preparing the theaflavin composition. On the other hand, both vitamin C and vitamin E have antioxidant activity. Vitamin C can rapidly decompose ethanol and convert it into acetaldehyde, accelerating alcohol excretion and exerting a hangover-relieving effect. Vitamin E can prevent drunkenness and also improve symptoms such as headache, dizziness, and stomach discomfort caused by alcohol.

[0023] The present invention also provides a method for preparing the theaflavins composition, comprising: mixing antioxidants, theaflavins, kudzu root extract, hovenia dulcis fruit extract, snow chrysanthemum extract, kelp extract and liquorice extract to obtain the theaflavins composition.

[0024] The present invention also provides the use of the theaflavin composition in preparing a drug for treating liver damage, wherein the liver damage is alcoholic liver damage.

[0025] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0026] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions of the present invention. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0027] In the following examples, unless otherwise specified, all methods are conventional.

[0028] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0029] Lactobacillus hilgensis GZ2 was purchased from the China General Microorganism Collection Center with a collection number of CGMCC NO. 26755, which is disclosed in patent CN116814468A. Bifidobacterium bifidum CICC 6169, Lactobacillus delbrueckii CICC 6289, and Bifidobacterium longum CICC 6197 were purchased from the China Industrial Microorganism Culture Collection Center. Kelp powder was purchased from Xinghua Mingwang Food Co., Ltd. Theaflavin was prepared according to Example 4 of invention patent CN117867052A, and the theaflavin content was 63.9% as determined by HPLC.

[0030] Example 1 Theaflavin composition (1) Preparation of Pueraria root extract The kudzu root was crushed, passed through a 500-mesh sieve, mixed with purified water at a solid-liquid ratio of 1:10 g / mL, inoculated with a composite bacterial agent, and subjected to liquid anaerobic fermentation at 37°C and a pH value of 5.5 for 60 hours. The mixture was sterilized, filtered, and the filtrate was concentrated and dried to a water content of 3 wt% to obtain a kudzu root extract. The composite bacterial agent is composed of Lactobacillus hilgeni GZ2 and Bifidobacterium bifidum CICC 6169. The bacterial count of Lactobacillus hilgeni GZ2 is 800 million / g, and the bacterial count of Bifidobacterium bifidum CICC 6169 is 300 million / g. The inoculation amount of the composite bacterial agent is 2% of the mass of Pueraria root. (2) Preparation of Hovenia dulcis extract The Hovenia dulcis fruit was crushed and passed through a 400-mesh sieve. A 70% by volume ethanol solution was added at a solid-liquid ratio of 1:10. Ultrasonic extraction was performed at a temperature of 50°C, a power of 300W, and a frequency of 25kHz for 30 minutes. The mixture was filtered, concentrated, and dried to a water content of 3wt% to obtain a Hovenia dulcis fruit extract. (3) Preparation of snow chrysanthemum extract The snow chrysanthemum was crushed, and a 60% volume fraction ethanol solution was added at a solid-liquid ratio of 1:12 g / mL. The mixture was ultrasonically extracted at a temperature of 45°C, a power of 230 W, and a frequency of 28 kHz for 35 min. The mixture was filtered, concentrated, and dried to a water content of 2 wt% to obtain a snow chrysanthemum extract. (4) Preparation of licorice extract The licorice was crushed, and a 50% volume fraction ethanol solution was added at a solid-liquid ratio of 1:18 g / mL. The mixture was ultrasonically extracted at a temperature of 42°C, a power of 280 W, and a frequency of 32 kHz for 45 min. The mixture was filtered, concentrated, and dried to a water content of 4 wt% to obtain a licorice extract. (5) Preparation of kelp extract The kelp powder and water were mixed at a solid-liquid ratio of 1:25 g / mL, and high-voltage pulsed electric field extraction was performed for 45 seconds at an electric field intensity of 25 kV / cm, a pulse width of 8 μs, and a temperature of 75°C. The filtrate was filtered, and the filtrate was concentrated to 1 / 3 of the volume of the supernatant. A 10-fold volume of a 95% ethanol solution was added, and the mixture was precipitated for 12 hours. The mixture was centrifuged, and the precipitate was dried to a water content of 6 wt% to obtain a kelp extract. (6) Preparation of theaflavins composition Accurately weigh 25 parts of theaflavins, 25 parts of kudzu root extract, 20 parts of Hovenia dulcis extract, 15 parts of snow chrysanthemum extract, 8 parts of kelp extract, 5 parts of licorice extract, and 0.02 parts of vitamin C; The above raw materials were passed through an 80-mesh sieve, and vitamin C, theaflavins, kudzu root extract, hovenia dulcis fruit extract, snow chrysanthemum extract, kelp extract and liquorice extract were mixed in order at a rotation speed of 80 rpm for 30 minutes to obtain the theaflavins composition.

[0031] Example 2 Theaflavin composition (1) Preparation of Pueraria root extract The kudzu root was crushed, passed through a 400-mesh sieve, mixed with purified water at a solid-liquid ratio of 1:8 g / mL, inoculated with a composite bacterial agent, and subjected to liquid anaerobic fermentation at 35°C and pH 5 for 72 hours, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 4 wt% to obtain a kudzu root extract; The composite bacterial agent is composed of Lactobacillus hilgeni GZ2 and Bifidobacterium bifidum CICC 6169. The bacterial count of Lactobacillus hilgeni GZ2 is 500 million / g, and the bacterial count of Bifidobacterium bifidum CICC 6169 is 200 million / g. The inoculation amount of the composite bacterial agent is 3% of the mass of Pueraria root. (2) Preparation of Hovenia dulcis extract The Hovenia dulcis fruit was crushed and passed through a 350-mesh sieve. A 65% volume fraction ethanol solution was added at a solid-liquid ratio of 1:8 g / mL. Ultrasonic extraction was performed at a temperature of 48°C, a power of 250 W, and a frequency of 23 kHz for 35 min. The extract was filtered, concentrated, and dried to a water content of 4 wt% to obtain the Hovenia dulcis fruit extract. (3) Preparation of snow chrysanthemum extract The snow chrysanthemum was crushed, and a 55% volume fraction ethanol solution was added at a solid-liquid ratio of 1:10 g / mL. The mixture was ultrasonically extracted at a temperature of 43°C, a power of 200 W, and a frequency of 25 kHz for 40 min. The mixture was filtered, concentrated, and dried to a water content of 4 wt% to obtain a snow chrysanthemum extract. (4) Preparation of licorice extract The licorice was crushed, and a 48% volume fraction ethanol solution was added at a solid-liquid ratio of 1:15 g / mL. The mixture was ultrasonically extracted at a temperature of 38°C, a power of 250 W, and a frequency of 30 kHz for 50 min. The mixture was filtered, concentrated, and dried to a water content of 3 wt% to obtain a licorice extract. (5) Preparation of kelp extract The kelp powder and water were mixed at a solid-liquid ratio of 1:20 g / mL, and high-voltage pulsed electric field extraction was performed for 60 s at an electric field intensity of 20 kV / cm, a pulse width of 5 μs, and a temperature of 70°C. The filtrate was filtered, and the filtrate was concentrated to 1 / 2 of the volume of the supernatant. An 8-fold volume of a 90% ethanol solution was added, and the mixture was precipitated for 14 h. The mixture was centrifuged, and the precipitate was dried to a water content of 3 wt% to obtain a kelp extract. (6) Preparation of theaflavins composition Accurately weigh 20 parts of theaflavins, 20 parts of kudzu root extract, 15 parts of Hovenia dulcis extract, 10 parts of snow chrysanthemum extract, 5 parts of kelp extract, 3 parts of licorice extract and 0.01 parts of vitamin E The above raw materials were passed through a 50-mesh sieve, and vitamin E, theaflavins, kudzu root extract, hovenia dulcis fruit extract, snow chrysanthemum extract, kelp extract and liquorice extract were mixed in sequence at a rotation speed of 60 rpm for 35 minutes to obtain the theaflavins composition.

[0032] Example 3 Theaflavin composition (1) Preparation of Pueraria root extract The kudzu root was crushed, passed through a 600-mesh sieve, mixed with purified water at a solid-liquid ratio of 1:12 g / mL, inoculated with a composite bacterial agent, and subjected to liquid anaerobic fermentation at 40°C and pH 6 for 58 hours, sterilized, filtered, and the filtrate was concentrated and dried to a water content of 5 wt% to obtain a kudzu root extract; The composite bacterial agent is composed of Lactobacillus hilgeni GZ2 and Bifidobacterium bifidum CICC 6169. The bacterial count of Lactobacillus hilgeni GZ2 is 1 billion / g, and the bacterial count of Bifidobacterium bifidum CICC 6169 is 500 million / g. The inoculation amount of the composite bacterial agent is 1% of the mass of Pueraria root. (2) Preparation of Hovenia dulcis extract The Hovenia dulcis fruit was crushed and passed through a 500-mesh sieve. A 75% volume fraction ethanol solution was added at a solid-liquid ratio of 1:12. Ultrasonic extraction was performed at a temperature of 52°C, a power of 350W, and a frequency of 28kHz for 25 minutes. The mixture was filtered, concentrated, and dried to a water content of 4wt% to obtain a Hovenia dulcis fruit extract. (3) Preparation of snow chrysanthemum extract The snow chrysanthemum was crushed, and a 65% volume fraction ethanol solution was added at a solid-liquid ratio of 1:15 g / mL. The mixture was ultrasonically extracted at a temperature of 48°C, a power of 250 W, and a frequency of 30 kHz for 30 min. The mixture was filtered, concentrated, and dried to a water content of 6 wt% to obtain a snow chrysanthemum extract. (4) Preparation of licorice extract The licorice was crushed, and a 53% volume fraction ethanol solution was added at a solid-liquid ratio of 1:20 g / mL. The mixture was ultrasonically extracted at a temperature of 45°C, a power of 300 W, and a frequency of 35 kHz for 40 min. The mixture was filtered, concentrated, and dried to a water content of 2 wt% to obtain a licorice extract. (5) Preparation of kelp extract The kelp powder and water were mixed at a solid-liquid ratio of 1:30 g / mL, and high-voltage pulsed electric field extraction was performed for 30 s at an electric field intensity of 30 kV / cm, a pulse width of 10 μs, and a temperature of 80°C. The filtrate was filtered, and the filtrate was concentrated to 1 / 4 of the volume of the supernatant. 12 times the volume of a 97% ethanol solution was added, and the mixture was precipitated for 10 h. The mixture was centrifuged, and the precipitate was dried to a water content of 4 wt% to obtain a kelp extract. (6) Preparation of theaflavins composition Theaflavins 30 parts, Pueraria root extract 30 parts, Hovenia dulcis extract 25 parts, Snow chrysanthemum extract 20 parts, Kelp extract 10 parts, Licorice extract 8 parts and Vitamin C 0.03 parts; The raw materials were passed through a 100-mesh sieve, and vitamin C, theaflavins, kudzu root extract, hovenia dulcis fruit extract, snow chrysanthemum extract, kelp extract and liquorice extract were mixed in sequence at a rotation speed of 100 rpm for 20 minutes to obtain the theaflavins composition.

[0033] Comparative Example 1 The specific implementation method is the same as that of Example 1, except that "25 parts of theaflavins and 25 parts of Pueraria root extract" in step (6) are replaced with "50 parts of theaflavins" and step (1) is deleted.

[0034] Comparative Example 2 The specific implementation method is the same as that of Example 1, except that "25 parts of theaflavins and 25 parts of Pueraria root extract" in step (6) are replaced with "50 parts of Pueraria root extract".

[0035] Comparative Example 3 The specific implementation method is the same as that of Example 1, except that the composite bacterial agent in step (1) is composed of Lactobacillus delbrueckii CICC6289 and Bifidobacterium longum CICC 6197, the bacterial count of Lactobacillus delbrueckii CICC 6289 in the composite bacterial agent is 800 million cells / g, the bacterial count of Bifidobacterium longum CICC 6197 is 300 million cells / g, the inoculation amount of the composite bacterial agent is 2% of the mass of Pueraria lobata, and the liquid anaerobic fermentation conditions remain unchanged.

[0036] Comparative Example 4 The specific implementation is the same as that of Example 1, except that the ethanol solutions in steps (2) to (4) are replaced with pure water.

[0037] Comparative Example 5 The specific implementation method is the same as that of Example 1, except that the preparation method of the kelp extract in step (5) is as follows: kelp powder and water are mixed at a solid-liquid ratio of 1:25 g / mL, heated and extracted at 85°C for 3 h, filtered, and the filtrate is concentrated to 1 / 3 of the volume of the supernatant, 10 times the volume of 95% ethanol solution is added, precipitated for 12 h, centrifuged, and the precipitate is dried to a water content of 6 wt% to obtain the kelp extract.

[0038] Test Example 1 Take before drinking SPF-grade 20±2g C57BL / 6 male mice were selected and divided into 11 groups, 10 mice in each group, including a blank control group, a positive control group, a model group, and an experimental group (Examples 1-3, Comparative Examples 1-5).

[0039] Adaptive feeding was performed for 7 days with free access to water and food. The temperature was maintained at 23-25°C, with a 12-hour day-night cycle. After 7 days of adaptive feeding, the positive control group was gavaged daily with 150 mg / kg of silymarin, the experimental groups were gavaged with 150 mg / kg of the corresponding theaflavin combination, and the blank control and model groups were gavaged with an equal volume of normal saline. After 30 minutes of gavage, a 52% ethanol solution was gavaged at 10 mL / kg. This gavage was continued for 15 days.

[0040] (1) Serum biochemical index detection On the last day of gavage, 6 h after gavage with ethanol solution, mice were anesthetized with ether, their eyes were removed and blood was collected. The blood was allowed to stand at room temperature and centrifuged at 4°C and 3000 rpm for 10 min. The upper layer was serum.

[0041] The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured according to the kit instructions, and the results are shown in Table 1.

[0042] Table 1 Effects of each group on serum ALT and AST levels Grouping ALT(U / L) AST(U / L) Blank control group 37.23±5.62 78.35±18.97 Model Group 116.77±20.13 217.81±64.23 Positive control group 45.59±4.95 105.55±23.40 Example 1 46.35±2.88 107.92±20.61 Example 2 47.17±3.30 109.37±22.14 Example 3 47.21±3.54 108.19±25.89 Comparative Example 1 72.84±7.07 137.03±36.06 Comparative Example 2 68.38±6.59 135.76±31.73 Comparative Example 3 65.90±6.71 126.60±24.02 Comparative Example 4 62.16±5.26 119.48±26.38 Comparative Example 5 53.42±5.43 112.24±27.55 As shown in Table 1, ALT and AST levels were significantly elevated in the model group compared to the blank control group, indicating successful modeling of alcoholic liver injury in mice. There was little difference in ALT and AST between the Example group and the blank control group, indicating that the theaflavins composition in the Example group has a protective effect against alcoholic liver injury. A comparison of Example 1 and Comparative Examples 1-2 demonstrates that theaflavins and Pueraria extract exhibit a synergistic effect, significantly reducing ALT and AST levels when used together. As shown in Example 1 and Comparative Examples 2-5, the protective effect of the theaflavins composition against alcoholic liver injury can be affected by using different microbial fermentation methods, different solvents for extraction, and different extraction methods.

[0043] (2) Liver biochemical index detection After blood collection, mice were killed by cervical dislocation. Liver tissue was dissected and liver mass was measured to calculate the liver index. Superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and malondialdehyde (MDA) content in the livers of mice in each group were measured using kits. The results are shown in Table 2.

[0044] Table 2 Effects of each group on liver index, SOD, GSH-Px and MDA in the liver Grouping Liver index SOD (U / mg) GSH-Px (U / mg) MDA (nmol / mg) Blank control group 4.47±0.47 136.02±32.27 551.12±98.50 6.59±0.82 Model Group 8.79±1.26 53.75±16.64 376.87±76.89 39.04±6.36 Positive control group 4.53±0.39 82.31±22.31 539.51±94.36 18.42±3.51 Example 1 4.67±0.43 80.84±17.05 535.30±82.74 20.85±4.18 Example 2 4.71±0.55 77.42±20.89 528.14±78.17 22.98±3.85 Example 3 4.75±0.41 79.18±22.40 533.79±96.82 22.06±4.07 Comparative Example 1 6.60±1.10 60.93±17.16 418.06±69.05 30.31±5.69 Comparative Example 2 6.56±1.28 61.66±16.72 434.93±63.61 31.67±5.24 Comparative Example 3 6.28±0.74 63.09±19.57 480.78±70.48 28.12±4.40 Comparative Example 4 5.84±0.52 70.22±18.93 461.43±75.93 26.78±2.93 Comparative Example 5 5.52±0.64 72.57±23.48 502.65±81.23 25.23±4.75 As shown in Table 2, the liver index in the model group was significantly higher than that in the blank control group, indicating that the model of alcoholic liver injury in mice was successfully established. The SOD, GSH-Px, and MDA data in the model and blank control groups indicate that SOD and GSH-Px activities were significantly reduced, while MDA content was increased, indicating that the liver damage in mice administered alcohol was severe. There was little difference in SOD, GSH-Px, and MDA levels between the Example group and the blank control group, indicating that the theaflavins composition in the Example group has a protective effect against alcoholic liver injury. A comparison of the data in Example 1 and Comparative Examples 1-2 shows that theaflavins and Pueraria extract have a synergistic effect, and their combined use can significantly increase SOD and GSH-Px activities and reduce MDA content. As shown in Example 1 and Comparative Examples 2-5, the use of different microbial fermentations, different solvent extractions, and different extraction methods can affect SOD, GSH-Px, and MDA levels, thereby affecting the protective effect of the theaflavins composition against alcoholic liver injury.

[0045] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A theaflavin composition for treating alcoholic liver damage, characterized in that: The invention comprises the following raw materials in parts by weight: 20-30 parts of theaflavins, 20-30 parts of kudzu root extract, 15-25 parts of hovenia dulcis fruit extract, 10-20 parts of snow chrysanthemum extract, 5-10 parts of kelp extract, 3-8 parts of liquorice extract and 0.01-0.03 parts of antioxidant.

2. The theaflavin composition according to claim 1, characterized in that The preparation method of the kudzu root extract comprises the following steps: crushing the kudzu root, mixing the kudzu root with water, inoculating a composite bacterial agent for liquid anaerobic fermentation, sterilizing, filtering, concentrating the filtrate, and drying to obtain the kudzu root extract.

3. The theaflavin composition according to claim 2, characterized in that The composite bacterial agent consists of Lactobacillus hilgris GZ2 and Bifidobacterium bifidum CICC 6169.

4. The theaflavin composition according to claim 2, characterized in that The temperature of the liquid fermentation is 35-40° C., the pH value is 5-6, and the fermentation time is 58-72 hours.

5. The theaflavin composition according to claim 1, characterized in that The Hovenia dulcis fruit extract, snow chrysanthemum extract and liquorice extract are obtained by extraction with ethanol solution.

6. The theaflavin composition according to claim 1, characterized in that The kelp extract is extracted by a high-voltage pulse electric field method.

7. The theaflavin composition according to claim 1, characterized in that The antioxidant is vitamin C or vitamin E.

8. A method for preparing the theaflavin composition according to any one of claims 1 to 7, characterized in that: include: The theaflavins composition is obtained by mixing antioxidants, theaflavins, kudzu root extract, hovenia dulcis fruit extract, snow chrysanthemum extract, kelp extract and liquorice extract.

9. Use of the theaflavin composition according to any one of claims 1 to 7 in the preparation of a drug for treating liver damage.

10. The use according to claim 9, characterized in that The liver damage is alcoholic liver damage.