A preparation method and application of a traditional Chinese medicine extract and trace element health food for improving alcoholic liver injury

Abstract

A method for preparing a health food product containing traditional Chinese medicine extract and trace elements for improving alcoholic liver injury, and its application. Through extraction, purification, freeze-drying, content determination, dosage form preparation, functional evaluation of the formulation, animal experiments, biochemical index analysis, toxicological studies, and research on the protective effect of the drug on alcohol-damaged cells, this traditional Chinese medicine health food product was verified to have the function of improving alcoholic liver injury. It improves alcoholic liver injury through oral absorption and effectively reduces clinical indicators such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), and triglycerides.

Description

Technical Field

[0001] This invention relates to a health food composition, specifically to a method for preparing and applying a health food that improves alcoholic liver damage, belonging to the field of health food research, production, and use. Background Technology

[0002] This invention is based on the theories of Traditional Chinese Medicine and, in accordance with the "Regulations for the Registration and Management of Health Foods," develops a health food composition and its preparation method that improves alcoholic liver damage. The specific ingredients are derived from commonly used medicinal and edible plants and trace elements required by the human body.

[0003] Alcoholic liver disease, commonly known as alcoholic hepatitis, is a disease caused by long-term excessive alcohol consumption, resulting in abnormal liver cell structure and / or dysfunction. Initially, it manifests as fatty liver with mild or no symptoms, or nonspecific symptoms such as right upper quadrant abdominal distension and pain, loss of appetite, fatigue, weight loss, and jaundice. As the condition worsens, it progresses to alcoholic hepatitis, alcoholic liver fibrosis, and alcoholic cirrhosis, and may present with neuropsychiatric symptoms, spider angiomas, and palmar erythema. Severe alcohol abuse can induce widespread hepatocellular necrosis and even liver failure. Therefore, further strengthening research on alcoholic liver injury and actively carrying out prevention and treatment of alcoholic liver injury and its complications is an urgent task facing the medical community. Traditional Chinese medicine has a long history of improving liver damage and has accumulated a wealth of valuable experience. In particular, the active ingredients derived from traditional Chinese medicine for treating liver diseases have the characteristics of mild and long-lasting therapeutic effects, few side effects, and long-term use. Furthermore, their mechanism of action often involves multi-target, multi-effect, and multi-functional comprehensive effects, possessing comprehensive advantages that Western medicine cannot replace.

[0004] This product's formula is based on the dosage of drugs in the Chinese Pharmacopoeia, with appropriate additions and subtractions. It is composed of medicinal and edible plants such as kudzu root, bitter orange seed, white poria, galangal, milk thistle, vitamin C, selenium-enriched yeast, calcium gluconate, and zinc citrate, as well as trace elements.

[0005] The formula contains kudzu root, immature bitter orange seeds, white poria cocos, and galangal, which are both food and medicine, while milk thistle is an ingredient that can be used in health food products. All the medicinal ingredients in the formula meet the requirements for raw material sources for health food products.

[0006] Kudzu root: The dried root of *Pueraria lobata* (willd.) ohwi, a legume, is harvested in spring and autumn, impurities are removed, and it is sun-dried. It is sweet, pungent, and cool in nature. It enters the spleen and stomach meridians. It is a commonly used herb in Traditional Chinese Medicine for dispelling wind and releasing the exterior, and has pharmacological effects such as lowering blood sugar, anti-tumor activity, antipyretic effect, dilating cerebral blood vessels, lowering blood pressure, slowing heart rate, reducing myocardial oxygen consumption, and relaxing smooth muscle.

[0007] Trifoliate orange (Hovenia dulcis Thumb): A woody plant belonging to the Rhamnaceae family and the Hovenia genus. In traditional Chinese medicine, it is believed to have the effects of clearing heat, promoting diuresis, and detoxifying alcohol. It is mainly used to treat symptoms such as drunkenness, fever, thirst, vomiting, and difficulty in urination and defecation. Its fruit axis can be used to treat rheumatism; the fruit stalk can strengthen the stomach and replenish blood, and is used to nourish and replenish blood; the leaves are used to treat stillbirths; and the leaf sap is used to eliminate body odor.

[0008] White Poria: The dried sclerotium of the fungus *Poria cocos* (Schw.) Wolf (family Polyporaceae). It is the white part of the medicinal Poria cocos after the red Poria cocos has been removed. It has the effects of improving diuresis and promoting urination; strengthening the spleen and stomach; and calming the mind and soothing the nerves. It is used for symptoms such as difficulty urinating; edema and abdominal distension; phlegm and cough; vomiting; poor appetite due to spleen deficiency; diarrhea; palpitations; insomnia and forgetfulness.

[0009] Galangal: The dried rhizome of Alpinia officinarum Hance, a plant in the ginger family. It is used for abdominal pain due to cold, vomiting due to stomach cold, belching and acid reflux, warming the stomach and stopping vomiting, and dispelling cold and relieving pain.

[0010] Milk thistle: A species of annual or biennial herb belonging to the genus *Silybum marianum* (L.) Gaertn. of the Asteraceae family. The entire plant contains fumaric acid and flavonoids, and the seeds are rich in silymarin (a collective term for silymarin A, B, silymarinine, silymarinine, etc.). Silymarin has heat-clearing and detoxifying effects, as well as liver-soothing and bile-promoting effects. It is commonly used for damp-heat in the liver and gallbladder, hypochondriac pain, and jaundice. In addition, milk thistle also has anti-inflammatory, anti-cancer, and cardiovascular protective effects.

[0011] Vitamin C: Vitamin C is a water-soluble vitamin, chemically named L-(+)-threose-type 2,3,4,5,6-pentahydroxy-2-hexenoic acid-4-lactone, also known as L-ascorbic acid, with the molecular formula C6H8O6 and a molecular weight of 176.12. It is odorless, has a sour taste, is easily soluble in water, and has strong reducing properties. It participates in complex metabolic processes in the body, promotes growth, and enhances resistance to disease. It can be used as a nutritional supplement and antioxidant.

[0012] Selenium-enriched yeast: Selenium, with atomic number 34 and relative atomic mass 78.96, is an essential trace element for the human body, playing a vital physiological role and closely related to human health. Selenium can prevent and inhibit tumors, combat aging, maintain the normal function of the cardiovascular system, and prevent arteriosclerosis and coronary heart disease. Furthermore, selenium is known as an "anti-liver necrosis protective factor," acting as one of the factors that prevent liver necrosis. Selenium protects liver cell membranes, thereby protecting the liver.

[0013] Calcium gluconate: Calcium gluconate is an organic calcium salt with the chemical formula C12H22O14Ca. As a medicine, it can reduce capillary permeability, increase calcium density, maintain normal nerve and muscle excitability, strengthen myocardial contractility, and aid in bone formation. Numerous studies have shown that excessive alcohol consumption leads to a significant loss of calcium from the body, which should be replenished promptly. As an organic calcium source, calcium gluconate is more easily absorbed by the body.

[0014] Zinc citrate: an organic compound with the chemical formula C12H14O16Zn3, mainly used as a zinc supplement, which has the effects of promoting digestion, enhancing appetite, and improving immunity.

[0015] Currently, no health food products composed of traditional Chinese medicine extracts and trace elements are available on the market. A search of Chinese patent and literature databases revealed no health food patents or research reports with similar formulations to the composition of this invention. The design theory and approach of this invention are innovative. Based on this invention, health food products with the function of improving alcoholic liver damage can be developed, with broad market prospects. Summary of the Invention

[0016] The present invention provides a health food composition based on traditional Chinese medicine health preservation theory, which improves alcoholic liver damage. It is prepared from natural raw materials and excipients that comply with the "Regulations for the Registration and Management of Health Foods". The present invention also provides a method for preparing this compound health food composition and its uses.

[0017] This invention provides a traditional Chinese medicine health food composition for improving alcoholic liver damage, which is a preparation made from the following raw materials in the indicated weight ratios:

[0018] 1-15 parts of kudzu root, 1-15 parts of bitter orange, 1-8 parts of white poria, 1-5 parts of galangal, 1-5 parts of milk thistle, 1-5 parts of vitamin C, 0.1-5 parts of selenium-enriched yeast, 1-5 parts of calcium gluconate, and 1-5 parts of zinc citrate.

[0019] More preferably, it is a formulation made from raw materials in the following weight ratios:

[0020] 15 parts kudzu root, 15 parts bitter orange, 8 parts white poria, 5 parts galangal, 5 parts milk thistle, 1 part vitamin C, 0.1 part selenium-enriched yeast, 1 part calcium gluconate, and 1 part zinc citrate.

[0021] The health food composition of the present invention is a preparation made from kudzu root, bitter orange seed, white poria, galangal, milk thistle, vitamin C, selenium-enriched yeast, calcium gluconate, zinc citrate and trace elements as active ingredients, plus excipients or auxiliary ingredients acceptable for health foods.

[0022] The dosage forms mentioned include: pills, powders, granules, tablets, chewable tablets, capsules, soft capsules, oral liquids, pastes, concentrated liquids, ointments, soups, cakes, and beverages.

[0023] The present invention also provides a method for preparing the health food composition, which includes the following steps:

[0024] (1) Weigh the raw materials according to the following weight ratio: 1-15 parts of kudzu root, 1-15 parts of bitter orange, 1-8 parts of white poria, 1-5 parts of galangal, and 1-5 parts of milk thistle.

[0025] (2) Take kudzu root, bitter orange seed, white poria, galangal, and milk thistle and extract them with water or organic solvent to obtain the extract;

[0026] (3) Take the extract and trace elements and add them to the excipients or auxiliary ingredients that are acceptable in health food, and make them into preparations according to the corresponding molding process.

[0027] This invention relates to a health food composition based on traditional Chinese medicine theory and empirical formulas used in clinical practice to improve alcoholic liver damage. The composition selects ingredients that meet the requirements for health food raw materials, with specific components derived from commonly used medicinal and edible plants. This achieves the goal of improving alcoholic liver damage and provides a new option for developing health foods. Detailed Implementation

[0028] The present invention will be described in more detail below with reference to experimental examples and embodiments.

[0029] Example 1

[0030] The health food of the present invention is made from the following ingredients by weight: 150g of kudzu root, 150g of bitter orange seed, 80g of white poria cocos, 50g of galangal, 50g of milk thistle, 10g of vitamin C, 1g of selenium-enriched yeast, 10g of calcium gluconate, and 10g of zinc citrate.

[0031] Furthermore, the health food of the present invention is prepared through the following stages:

[0032] (1) Take the above weight of kudzu root, bitter orange seed, white poria, galangal and milk thistle, add water and decoct twice, filter, combine the filtrates to obtain the extract, the extract is purified by adsorption clarification or ultracentrifugation to obtain the purified liquid, the purified liquid is concentrated under reduced pressure to obtain the extract for use.

[0033] (2) Take appropriate excipients and mix with the above extract, dry, pulverize, pass through a 100-mesh sieve, add trace elements, use 50% ethanol as a wetting agent to make soft material, extrude and pass through a 14-mesh sieve to granulate, dry, pass through a 16-mesh sieve to granulate, and obtain granules.

[0034] Example 2

[0035] The health food of the present invention is made from the following ingredients by weight: 150g of kudzu root, 150g of bitter orange seed, 80g of white poria cocos, 50g of galangal, 50g of milk thistle, 10g of vitamin C, 1g of selenium-enriched yeast, 10g of calcium gluconate, and 10g of zinc citrate.

[0036] Furthermore, the health food of the present invention is prepared through the following stages:

[0037] (1) Take the above weight of kudzu root, bitter orange seed, white poria, galangal and milk thistle, add water and decoct twice, filter, combine the filtrates to obtain the extract, the extract is purified by adsorption clarification or ultracentrifugation to obtain the purified liquid, the purified liquid is concentrated under reduced pressure to obtain the extract for use.

[0038] (2) Dry, pulverize, pass through a 100-mesh sieve, add trace elements, use 50% ethanol as a wetting agent to make soft material, extrude through a 14-mesh sieve to granulate, dry, pass through a 16-mesh sieve to granulate, obtain dry granules, use a tablet press to compress the dry granules into tablets, and obtain chewable tablets.

[0039] Example 3

[0040] The health food of the present invention is made from the following ingredients by weight: 150g of kudzu root, 150g of bitter orange seed, 80g of white poria cocos, 50g of galangal, 50g of milk thistle, 10g of vitamin C, 1g of selenium-enriched yeast, 10g of calcium gluconate, and 10g of zinc citrate.

[0041] Furthermore, the health food of the present invention is prepared through the following stages:

[0042] (1) Take the above weight of kudzu root, bitter orange seed, white poria, galangal and milk thistle, add water and decoct twice, filter, combine the filtrates to obtain the extract, the extract is purified by adsorption clarification or ultracentrifugation to obtain the purified liquid, the purified liquid is concentrated under reduced pressure to obtain the concentrated liquid for use.

[0043] (2) Take 100g of agar and put it into 20 times the amount of concentrated solution. Soak for 10 hours and heat until dissolved. Add trace elements, sweetener and natural preservative. Stir well, heat to 80℃, stir evenly, sterilize by hot pressing, cool to 80℃ and pour into mold to obtain the soup.

[0044] Implementation Example 4

[0045] The health food of the present invention is made from the following ingredients by weight: 150g of kudzu root, 150g of bitter orange seed, 80g of white poria cocos, 50g of galangal, 50g of milk thistle, 10g of vitamin C, 1g of selenium-enriched yeast, 10g of calcium gluconate, and 10g of zinc citrate.

[0046] Furthermore, the health food of the present invention is prepared through the following stages:

[0047] (1) Take the above weight of kudzu root, bitter orange seed, white poria, galangal and milk thistle, add water and decoct twice, filter, combine the filtrates to obtain the extract, and then refine the extract by adsorption clarification or ultracentrifugation to obtain the refined liquid for use.

[0048] (2) Take the above refined liquid, add trace elements, appropriate amount of flavoring agent and stabilizer, mix well, filter, add appropriate amount of pure water to dilute to the specified amount, mix well, fill, sterilize, and the beverage is obtained.

[0049] Experimental Example 1: Determination of total flavonoid content in the granules of this invention

[0050] The total flavonoid content in the preparation was determined by ultraviolet spectrophotometry.

[0051] 1. Solution preparation

[0052] 1.1 Sample Solution

[0053] Take the granules, crush them and pass them through a 60-mesh sieve. Accurately weigh 2g of the powder, add ethanol and sonicate for 30min. Make up to 25mL, shake well and filter through a microporous membrane to obtain the sample solution.

[0054] 1.2 Test solution

[0055] Accurately measure 4 mL of the sample solution and place it in an evaporating dish. Add 1 g of polyamide powder for adsorption. Evaporate the ethanol in a water bath or constant temperature oven. Then transfer it to a chromatography column. Wash with 30 mL of petroleum ether and discard the petroleum ether solution. Elute the flavonoids with methanol, collect the methanol solution, evaporate to dryness under reduced pressure, and then dissolve and dilute to 25 mL with methanol to obtain the final product.

[0056] 1.3 Reference Solution

[0057] Accurately weigh 5 mg of rutin reference standard, dissolve it in methanol, sonicate and cool, then dilute to 100 mL with methanol to obtain 50 μg / mL. -1 Reference solution.

[0058] 2. Examination of linear relationships

[0059] Take 0, 1, 2, 3, 4, and 5 mL of the reference solution respectively into 10 mL volumetric flasks, add methanol to the mark, shake well, and measure the absorbance at 360 nm. Perform linear regression between the concentration (X) of the reference solution and the absorbance value (Y), obtaining the regression equation Y = 0.0263X - 0.0485, r = 0.9992 (n = 6). The results show that within the range of 0-25 μg / mL... -1 Within the specified range, the concentration of the reference standard and the absorbance value showed a good linear relationship.

[0060] 3. Precision test

[0061] The reference solution was precisely measured and the measurement was repeated 5 times. The results are shown in Table 1, indicating that the instrument has good precision.

[0062] Table 1. Precision test results

[0063]

[0064] 4. Stability test

[0065] Accurately pipette the test solution and measure the absorbance at 0, 2, 4, 8, 12, and 24 hours. The results are shown in Table 2. The results indicate that the test solution is stable within 24 hours.

[0066] Table 2. Stability test results

[0067]

[0068] 5. Repeatability test

[0069] Accurately weigh the same batch of samples and prepare five test samples according to the test solution section, then measure the absorbance. The results are shown in Table 3. The results indicate that this method has good repeatability.

[0070] Table 3 Results of Repeatability Testing

[0071]

[0072] 6. Recovery rate study

[0073] Six aliquots of sample solution with known content were accurately measured, and appropriate amounts of reference standard were added to each. The solutions were prepared according to the procedure for test solutions, and the recovery rate was calculated. The results are shown in Table 4. The average recovery rate of this method was 95.14%, with an RSD of 1.42%, indicating that the method has good accuracy and is feasible.

[0074] Table 4 Results of the Recovery Rate Study

[0075]

[0076] 7. Sample content determination

[0077] The above-described determination method was used to determine the content of three batches of samples, and the results are shown in Table 5.

[0078] Table 5. Results of sample content determination

[0079]

[0080] Experimental Example 2: Determination of Puerarin Content in the Granules of the Present Invention

[0081] The content of puerarin in the preparation was determined by high performance liquid chromatography.

[0082] 1. Plotting the standard curve of puerarin

[0083] The content of puerarin in kudzu root extract was determined by reversed-phase high-performance liquid chromatography according to the pharmacopoeia.

[0084] Chromatographic column: Thermo Hypersil GOLD C18 column (250 mm × 4.6 mm, 5 μm) from the United States, with methanol-water (25:75) as the mobile phase, detection wavelength of 250 nm, flow rate of 1.0 ml / min; UV detection wavelength of 250 nm; injection volume of 10 μL, column temperature of 30 ℃ [2].

[0085] The results are as follows Figure 1 (See attached diagram in the instruction manual). Accurately weigh an appropriate amount of puerarin reference standard and prepare standard solutions of 0.1 mg / mL, 0.05 mg / mL, 0.025 mg / mL, 0.0125 mg / mL, and 0.00625 mg / mL puerarin using 30% ethanol. Perform high-performance liquid chromatography (HPLC) analysis under the specified chromatographic conditions. Based on the peak area (Y) and concentration (X), perform linear regression to obtain the linear equation: y = 88803x + 34.694R. 2 =0.9998, indicating that puerarin showed good linearity in the concentration range of 0.00625-0.1 mg / mL.

[0086] 2. Determination of puerarin content in kudzu root extract

[0087] According to the pharmacopoeia method, accurately weigh approximately 0.1 g of the extract powder (passed through a No. 3 sieve), place it in a stoppered conical flask, accurately add 50 ml of 30% ethanol, weigh, heat under reflux for 30 minutes, cool, weigh again, replenish the lost weight with 30% ethanol, shake well, filter, and collect the filtrate to obtain the test solution. Perform the determination under the above-described liquid chromatography conditions. The chromatogram is shown below. Figure 2 (See attached diagram in the instruction manual) The resulting extract powder contained 43.5% puerarin.

[0088] 3. Precision Experiment

[0089] Low (10 μg / ml), medium (50 μg / ml), and high (100 μg / ml) puerarin reference solutions were prepared using 30% ethanol. Under the chromatographic conditions described in section 2.1.1, each solution was repeatedly measured five times within the same day to determine the intra-day precision. The results are shown in Table 6, with RSD values ​​of 1.00%, 0.87%, and 0.38%, respectively. The same method was used to prepare three concentrations (low, medium, and high) of the solutions, and the samples were repeatedly injected within five days to determine the inter-day precision. The results are shown in Table 7, with RSD values ​​of 1.00%, 0.87%, and 0.38%, respectively.

[0090] Table 6. Intra-day precision of puerarin content determination

[0091]

[0092] Table 7. Daytime precision of puerarin content determination

[0093]

[0094] Functional evaluation and toxicological studies of the formulation in Experimental Example 3

[0095] 1. Functional evaluation experiment for improving alcoholic fatty liver

[0096] 1.1 Samples The samples are granules of this product. In this experiment, 5, 10, and 20 times the dosage were used as test substances.

[0097] 1.2 Adult animals were selected for the experiment. Mice (20±2g) of single sex were selected, with 10-15 mice in each group.

[0098] 1.3 Dosage Grouping and Test Sample Administration Time: The experiment included three dosage groups, one model control group, and one blank control group. One dosage group used 10 times the recommended human dose, and the other two dosage groups were also included.

[0099] 1.4 Establishment and Administration of Acute Alcoholic Liver Injury Model Except for the blank control group, the model control group received no drugs. Mice were administered 5 g / kg alcohol by gavage every 12 hours for three consecutive times to establish the acute alcoholic liver injury model. Simultaneously with model establishment, the composition was dissolved in physiological saline and administered by gavage for three consecutive times. After the last gavage, mice were anesthetized with ether, and blood was collected from the carotid artery.

[0100] 1.4.1 Effects of ALT and AST Levels in the Serum of Mice with Acute Alcoholic Model: Compared with the alcohol group, the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels in the normal group were significantly different (P<0.001), indicating that acute alcohol modeling can impair liver function in mice, with a significant increase in ALT and AST. Administration of different doses of the composition significantly reduced the serum ALT and AST levels in mice with liver injury (P<0.01). The high-dose group A showed a significant difference compared with the model group (P<0.01), indicating that this dosage significantly improved ALT and AST levels. This result demonstrates that the composition can inhibit the increase in liver transaminase levels in mice induced by acute alcohol. The results are as follows... Figure 3 , Figure 4 .

[0101] 1.4.2 Effect of the composition on serum TG levels in mice with acute alcoholic disease: Compared with the alcohol group, the serum triglyceride (TG) levels in the normal group were significantly different (P<0.001), indicating successful establishment of the acute alcohol model and a significant increase in TG. Administration of different doses of the composition significantly reduced serum TG levels in mice with liver injury (P<0.01). The high-dose composition group showed a significant difference compared with the model group (P<0.01), indicating that this dosage significantly improved TG levels. This result demonstrates that the composition can inhibit the increase in liver triglyceride levels in mice induced by acute alcohol. The results are as follows... Figure 5 .

[0102] 1.4.3A Effects on Liver Pathology in Acute Alcoholic Model Mice

[0103] The results are as follows Figure 6 (See attached diagram in the instruction manual). H&E staining results showed that the normal group had intact hepatocyte structure, arranged radially around the central vein, with no inflammatory cell infiltration in the portal area. Compared to the normal group, the acute alcohol-induced mouse hepatocytes were enlarged, the hepatic cords were disordered, the lobular structure was destroyed, and there was extensive inflammatory cell infiltration in and around the portal area. The cytoplasm was filled with fat vacuoles of varying sizes, and fatty degeneration was obvious, proving the successful establishment of an acute alcoholic mouse model. After administration of different doses of A, the degree of liver damage was significantly alleviated compared to the model group. The hepatic cords basically returned to their radial arrangement, and most of the hepatocyte structure returned to normal, with a small amount of inflammatory cell infiltration and tiny fat vacuoles visible.

[0104] 1.5 Data Processing and Result Determination

[0105] Experimental data were obtained using Graphpad Prism program 5.0 (Graphpad software, Inc., San Diego, USA). All measurements were Mean+SD. One-way ANOVA and Turkey's multivariate t-test were used to compare the data.

[0106] 1.6 Results

[0107] Figure 3 Serum ALT levels in each group of animals after drug administration. Compared with the normal group, #P<0.05, ##P<0.01, ###P<0.001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001.

[0108] Figure 4Serum AST levels in each group of animals after drug administration. Compared with the normal group, #P<0.05, ##P<0.01, ###P<0.001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001.

[0109] Figure 5 Serum TG levels in each group of animals after drug administration. Compared with the normal group, #P<0.05, ##P<0.01, ###P<0.001; compared with the model group, *P<0.05, **P<0.01, ***P<0.001.

[0110] 2. Toxicological Studies

[0111] The test results showed that the acute oral toxicity test of the granules of this invention in mice was effective. 50 The concentration was greater than 15.0 g / kg bw, classifying it as non-toxic according to toxicity classification. Micronucleus, sperm abnormality, and Ames tests in mice were all negative. A 30-day feeding experiment showed no adverse effects on complete blood count, blood biochemical indicators, organ coefficients, body weight, weight gain, food intake, or food utilization. Histopathological examination revealed no significant pathological changes in the liver, spleen, kidneys, stomach, duodenum, testes, or ovaries.

[0112] Experiment Example 4: Study on the Effects of Drugs on Cells

[0113] 1. Cytotoxicity assay of the drug

[0114] The effects of different concentrations of granules on the viability of HepG2 cells were detected using the CCK-8 assay.

[0115] The results are as follows Figure 7 (See attached diagram in the instruction manual). After reaching a cell density of 85%-90% in HepG2, cells were digested with trypsin and then seeded into 96-well plates (2*10⁴ cells / well) and cultured for 24 h. Cells were then cultured for another 24 h in serum-free medium containing different concentrations of the drug. After the culture period, 10 μl of CCK8 solution was added to each well. The absorbance at 450 nm was measured 1 h later. Cell viability was expressed as the ratio of absorbance between the treated and untreated groups.

[0116] 2. The protective effect of drugs on alcohol-damaged cells

[0117] After HepG2 cells reached a suitable density, 500 mM ethanol was added to each well for incubation for 24 hours. Then, drugs (100 μg / mL, 50 μg / mL, 25 μg / mL) were added, and the cells were incubated for another 24 hours. Cell viability was then detected using the CCK-8 assay.

[0118] 3 Results

[0119] The results are as follows Figure 8 (See attached diagram from the instruction manual). Compared to the control group, increased drug concentration had no significant effect on cell viability. The drug was non-toxic to HepG2 cells in the range of 6.25-200 μg / mL and can be used in subsequent experiments. In this study, 100 μg / mL, 50 μg / mL, and 25 μg / mL were selected as the concentrations for application to HepG2 cells.

[0120] Compared with the non-alcohol group, the cell viability of the alcohol-treated group was significantly reduced. Conversely, the relative cell viability of the drug-treated groups (100 μg / mL, 50 μg / mL, and 25 μg / mL) was significantly increased compared with the model group. The results indicate that the drug treatment groups in the concentration range of 25-100 μg / mL showed a significant protective effect against alcohol-damaged HepG2 cells.

[0121] The above functional evaluation experiments show that the health food of the present invention has the function of improving alcoholic liver damage and has no toxic side effects. Attached Figure Description

[0122] In the attached diagram:

[0123] Figure 1 Standard curve of puerarin

[0124] Figure 2 Graph showing the determination of puerarin content

[0125] Figure 3 Alanine aminotransferase (ALT) biochemical indicator

[0126] Figure 4 Aspartate aminotransferase (AST) biochemical indicator

[0127] Figure 5 For triglyceride (TG) biochemical indicators

[0128] Figure 6 Image of a liver pathological section

[0129] Figure 7 For drug cytotoxicity assay

[0130] Figure 8 Figure showing the protective effect of drugs on alcohol-damaged cells.

[0131] The scope of protection of this invention should not be considered limited to the specific embodiments and examples described above. For those skilled in the art, several simple deductions or equivalent substitutions can be made without departing from the basic concept of this invention, and these equivalent substitutions will still be considered within the scope of protection of this invention.

Claims

1. A method for preparing and applying a health food containing traditional Chinese medicine extracts and trace elements for improving alcoholic liver damage, comprising the following steps: (1) The raw materials of the Chinese herbal medicine extract were extracted using an ethanol aqueous solution with a mass fraction of 20-40% as a solvent, and the liquid portion was collected to obtain the extract. The raw materials of the Chinese herbal extract, by weight, consist of the following components: 2-4 parts of kudzu root, 2-4 parts of bitter orange seed, 2-4 parts of white poria, 2-4 parts of galangal, and 1-2 parts of milk thistle. (2) The extract is adsorbed by adsorption clarification method, and the liquid portion is collected to obtain a purified liquid; (3) The purified liquid was purified using a D101 macroporous resin column, the eluent was collected and dried to obtain the Chinese herbal extract; The purification conditions included: the eluent was a 25-35% (w / w) aqueous ethanol solution; the loading concentration was 0.05 g / ml; the loading volume was 6-7 BV; the flow rate was 1 BV / h; the eluent volume was 6-7 BV; and the elution flow rate was 1 BV / h.

2. The preparation method according to claim 1, characterized in that, The eluent is a 30% (w / w) aqueous solution of ethanol.

3. A traditional Chinese medicine extract for improving alcoholic liver injury obtained by the preparation method according to any one of claims 1 to 2, characterized in that, The total flavonoids in the herbal extract have a mass ratio of 5 to 15.

4. The traditional Chinese medicine extract for improving alcoholic liver injury according to claim 3, characterized in that, The total flavonoid content in the herbal extract is 6-8 mg / g.

5. A medicament for improving alcoholic liver injury, comprising the traditional Chinese medicine extract as described in claim 3 or 4 and pharmaceutical excipients.

6. The medicament for improving alcoholic liver injury according to claim 5, characterized in that, By weight percentage, it comprises 50-80% of the traditional Chinese medicine extract as described in claim 4 or 5, and 20-50% of pharmaceutical excipients.

7. The medicament for improving alcoholic liver injury according to claim 5 or 6, characterized in that, The pharmaceutical excipients include lactose, MCC, CMC-Na, magnesium stearate, and an aqueous solution of ethanol with a mass fraction of 60-80%.

8. The medicament for improving alcoholic liver injury according to claim 5, characterized in that, The dosage form of the drug is selected from granules, tablets, powders, capsules, oral liquids, injections, sprays, aerosols, powder inhalers, nasal drops, suppositories, pills, microspheres, or tinctures.

9. The medicament for improving alcoholic liver injury according to claim 8, characterized in that, When the drug dosage form is a tablet, it comprises the following components in parts by weight: 320 to 400 parts of the traditional Chinese medicine extract as described in claim 3 or 4, 60 to 80 parts of lactose, 40 to 60 parts of MCC, 20 to 30 parts of CMS-Na, 1 to 8 parts of magnesium stearate, and 0.1 to 1 part of an aqueous ethanol solution with a mass fraction of 60 to 80%.

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