Traditional Chinese medicine composition and lipidosome for dispelling effects of alcohol and protecting liver as well as preparation method and application of traditional Chinese medicine composition and lipidosome

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition and liposome for dispelling effects of alcohol and protecting liver as well as a preparation method and application thereof. The traditional Chinese medicine composition for dispelling the effects of alcohol and protecting the liver is prepared from the following raw materials in parts by mass: 1-100 parts of semen hoveniae, 1-100 parts of flos puerariae and 1-100 parts of radix puerariae. The hovenia dulcis thunb, the flower of kudzuvine and the root of kudzu vine are used as raw materials, and the hovenia dulcis thunb, the flower of kudzuvine and the root of kudzu vine are in synergistic interaction, so that the hovenia dulcis thunb, the flower of kudzuvine and the root of kudzu vine have the effects of dispelling effects of alcohol and protecting liver, have definite effects of improving liver functions and liver pathology on liver injury caused by ethanol, and are Furthermore, compared with common oral dosage forms, lipidosome, paste and patch prepared from the traditional Chinese medicine composition can bypass the first-pass effect of the liver, reduce decomposition of the medicine in the metabolic process, improve the effective medicine concentration and further improve the effects of dispelling the effects of alcohol and protecting the liver.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition and liposome for relieving alcohol and protecting the liver, and a preparation method and application thereof. Background Art

[0002] Alcoholic Liver Disease (ALD) is liver damage caused by long-term excessive alcohol consumption, mainly manifested as hepatocyte damage and inflammatory response. The course of ALD usually starts from fatty liver (accumulation of fat in hepatocytes), and can develop into alcoholic hepatitis with continuous drinking, and may lead to liver fibrosis or even cirrhosis in severe cases. Further deterioration of cirrhosis may cause liver failure or liver cancer. The symptoms of early ALD are relatively hidden, and symptoms such as fatigue, loss of appetite, jaundice, and liver pain may appear as the disease progresses.

[0003] The existing methods for treating ethanol (alcohol) poisoning mainly include drug therapy. However, drug therapy is relatively slow, has side effects and is prone to drug resistance, and the effect is very unsatisfactory. Traditional Chinese medicine, like food, comes from minerals, plants, and animal medicines in nature, has no toxic and side effects, and traditional Chinese medicine has no drug resistance and can be continuously used until the disease is cured. Therefore, it is particularly important to provide a traditional Chinese medicine composition for relieving alcohol and protecting the liver to treat alcohol poisoning in this field. Summary of the Invention

[0004] The purpose of the present invention is to provide a traditional Chinese medicine composition and liposome for relieving alcohol and protecting the liver, and a preparation method and application thereof, which have the effect of relieving alcohol and protecting the liver, have a definite effect on improving liver function and liver pathology caused by ethanol, and have no obvious effect on regulating blood lipid.

[0005] The present invention provides a traditional Chinese medicine composition for relieving alcohol and protecting the liver, comprising the following raw materials in parts by mass: 1-100 parts of Hovenia dulcis, 1-100 parts of Puerariae flos, and 1-100 parts of Puerariae lobatae radix.

[0006] Preferably, the dosage form of the traditional Chinese medicine composition includes a gel.

[0007] The present invention also provides a preparation method of the traditional Chinese medicine composition according to the above technical solution, comprising the following steps:

[0008] Soaking a mixture of Hovenia dulcis, Puerariae flos, and Puerariae lobatae radix with water to obtain a soaked product; the mass-volume ratio of the mixture to water is 1 g: 25-60 mL, the temperature of the water is 5-25 °C; the soaking time is 30-60 min;

[0009] Decocting the soaked product to obtain a decoction;

[0010] Mix the decoction and organic acid at 80 °C to obtain a traditional Chinese medicine composition.

[0011] Preferably, the decocting time is ≥ 30 min;

[0012] The mass-volume ratio of the organic acid to the decoction is 20 - 40 g: 1 L.

[0013] The present invention also provides the use of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition obtained by the preparation method described in the above technical solution in the preparation of products for relieving hangover and protecting the liver and / or treating alcoholic liver disease.

[0014] Preferably, the product includes a pharmaceutical preparation; the dosage form of the pharmaceutical preparation includes liposomes, ointments and patches.

[0015] The present invention also provides a liposome for relieving hangover and protecting the liver, comprising a traditional Chinese medicine composition and a thickening agent;

[0016] The traditional Chinese medicine composition is the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition obtained by the preparation method described in the above technical solution;

[0017] The thickening agent includes carboxymethyl cellulose and / or gelatin.

[0018] The present invention also provides a preparation method of the liposome described in the above technical solution, comprising the following steps: dispersing the traditional Chinese medicine composition into an ethanol solution with a volume concentration of 30 - 80% by ultrasonic treatment to obtain a dispersion; mixing the dispersion and the thickening agent to obtain the liposome.

[0019] The present invention also provides an ointment for relieving hangover and protecting the liver, comprising a liposome and a stabilizer;

[0020] The liposome is the liposome described in the above technical solution or the liposome obtained by the preparation method described in the above technical solution;

[0021] The stabilizer includes a suspending agent and / or a surfactant.

[0022] The present invention also provides a patch for relieving hangover and protecting the liver, comprising a breathable material and an ointment coated on the breathable material; the ointment is the ointment described in the above technical solution.

[0023] Beneficial effects:

[0024] The present invention provides a traditional Chinese medicine composition for relieving hangover and protecting the liver, comprising the following raw materials in parts by mass: 1-100 parts of Hovenia dulcis Thunb., 1-100 parts of Puerariae flos, and 1-100 parts of Pueraria lobata. The present invention uses Hovenia dulcis Thunb., Puerariae flos, and Pueraria lobata as raw materials. Hovenia dulcis Thunb. can protect the liver, prevent the formation of peroxidized lipids, enhance the motor function of the small intestine, reduce the absorption of ethanol by the small intestine, accelerate gastrointestinal motility, and promote the gastrointestinal emptying of ethanol; Puerariae flos can protect the enzyme activity of alcohol dehydrogenase and activate the microsomal ethanol oxidase pathway to accelerate the elimination rate of ethanol and its metabolites; puerarin in Pueraria lobata inhibits the absorption of ethanol by the digestive tract, strengthens the first-pass elimination of ethanol in the gastrointestinal tract, and Pueraria lobata can accelerate the metabolism rate of ethanol in the body by increasing the activity of alcohol dehydrogenase. The present invention combines Hovenia dulcis Thunb., Puerariae flos, and Pueraria lobata, and they have synergistic effects, with the efficacy of relieving hangover and protecting the liver. It has a clear effect on improving liver function and liver pathology caused by ethanol, and has no obvious effect on the regulation of blood lipids.

[0025] Furthermore, the present invention uses the above-mentioned traditional Chinese medicine composition to prepare liposomes, creams, and patches. Compared with common oral dosage forms, it can bypass the first-pass effect of the liver, reduce the decomposition of drugs during metabolism, increase the effective drug concentration, and further improve the efficacy of relieving hangover and protecting the liver. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required to be used in the embodiments.

[0027] Figure 1 It is the HE staining result of the livers of mice in each treatment group;

[0028] Figure 2 It is the Sirius red staining result of the livers of mice in each treatment group. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0029] The present invention provides a traditional Chinese medicine composition for relieving hangover and protecting the liver, comprising the following raw materials in parts by mass: 1-100 parts of Hovenia dulcis Thunb., 1-100 parts of Puerariae flos, and 1-100 parts of Pueraria lobata.

[0030] By mass parts, the raw materials of the traditional Chinese medicine composition of the present invention include 1 to 100 parts of Hovenia dulcis Thunb. As an embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 2 to 90 parts of Hovenia dulcis Thunb.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 3 to 80 parts of Hovenia dulcis Thunb.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 4 to 70 parts of Hovenia dulcis Thunb.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 5 to 60 parts of Hovenia dulcis Thunb.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 6 to 50 parts of Hovenia dulcis Thunb.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 8 to 40 parts of Hovenia dulcis Thunb.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 9 to 30 parts of Hovenia dulcis Thunb.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 10 parts of Hovenia dulcis Thunb. The present invention uses Hovenia dulcis Thunb. as the raw material. Chemical components in Hovenia dulcis Thunb., such as flavonoids and saponins, can improve the activities of SOD and glutathione peroxidase (GPX), which is beneficial to scavenging excessive free radicals generated in the body after drinking, preventing the formation of lipid peroxides, and protecting the liver; by increasing the level of nitric oxide (NO) to relax the smooth muscle of the small intestinal blood vessels, increasing the blood flow of the small intestinal mucosa, and increasing the material exchange rate of the small intestine, thereby enhancing the motor function of the small intestine, reducing the absorption of ethanol by the small intestine, and reducing the ethanol entering the blood circulation; at the same time, Hovenia dulcis Thunb. can accelerate gastrointestinal motility, promote the gastrointestinal emptying of ethanol, and achieve the purpose of relieving alcoholism.

[0031] Based on the mass parts of the Hovenia dulcis Thunb., the raw materials of the traditional Chinese medicine composition of the present invention include 1 to 100 parts of Pueraria flos. As an embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 2 to 90 parts of Pueraria flos.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 3 to 80 parts of Pueraria flos.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 4 to 70 parts of Pueraria flos.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 5 to 60 parts of Pueraria flos.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 6 to 50 parts of Pueraria flos.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 8 to 40 parts of Pueraria flos.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 9 to 30 parts of Pueraria flos.; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 10 parts of Pueraria flos. The present invention uses Pueraria flos. as the raw material. By protecting the gastric mucosa and inhibiting the absorption of ethanol by the digestive tract, reducing the blood ethanol concentration after drinking, protecting the enzyme activity of alcohol dehydrogenase, and activating the microsomal ethanol oxidase pathway, accelerating the elimination rate of ethanol and its metabolites, thereby achieving the purpose of relieving alcoholism; at the same time, the active ingredient of Pueraria flos., total flavonoids of Pueraria flos., can scavenge intracellular reactive oxygen species and free radicals, achieving an antioxidant effect.

[0032] Based on the mass parts of the Hovenia dulcis, the raw materials of the traditional Chinese medicine composition of the present invention include 1 to 100 parts of Pueraria lobata. As an embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 2 to 90 parts of Pueraria lobata; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 3 to 80 parts of Pueraria lobata; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 4 to 70 parts of Pueraria lobata; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 5 to 60 parts of Pueraria lobata; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 6 to 50 parts of Pueraria lobata; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 8 to 40 parts of Pueraria lobata; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 9 to 30 parts of Pueraria lobata; as another embodiment, the raw materials of the traditional Chinese medicine composition of the present invention include 10 parts of Pueraria lobata. The present invention uses Pueraria lobata as a raw material. Puerarin in Pueraria lobata inhibits the absorption of ethanol by the digestive tract, strengthens the first-pass elimination of ethanol in the gastrointestinal tract, and Pueraria lobata can accelerate the metabolism rate of ethanol in the body by increasing the activity of alcohol dehydrogenase. The crude polysaccharide and refined polysaccharide of Pueraria lobata have a scavenging effect on superoxide anion radicals, hydroxyl radicals and nitrite, achieving the purpose of relieving alcoholism.

[0033] The present invention combines Hovenia dulcis, Pueraria flos and Pueraria lobata. Hovenia dulcis, Pueraria flos and Pueraria lobata have synergistic effects, have the efficacy of relieving alcoholism and protecting the liver, have a definite effect on improving liver function and liver pathology caused by ethanol, and have no obvious effect on the regulation of blood lipid.

[0034] As an embodiment, the dosage form of the traditional Chinese medicine composition of the present invention includes a gel.

[0035] The present invention also provides a preparation method of the traditional Chinese medicine composition of the above technical solution, including the following steps:

[0036] Soak the mixture of Hovenia dulcis, Pueraria flos and Pueraria lobata with water to obtain the soaked product; the mass-volume ratio of the mixture and water is 1 g: 25 to 60 mL, the temperature of the water is 5 to 25 °C; the soaking time is 30 to 60 min;

[0037] Decoct the soaked product to obtain a decoction;

[0038] Mix the decoction and an organic acid at 80 °C to obtain the traditional Chinese medicine composition.

[0039] As an embodiment, the present invention mixes Hovenia dulcis, Pueraria flos and Pueraria lobata to obtain a mixture. The present invention has no strict requirements on the mixing method, and the conventional method in the art can be used for mixing. As an embodiment, the present invention does not need to crush Hovenia dulcis, Pueraria flos and Pueraria lobata, and can be directly mixed.

[0040] After obtaining the mixture, the present invention soaks the mixture with water to obtain the soaked product. As an embodiment, the mass-volume ratio of the mixture and water in the present invention is 1 g: 30-55 mL; as another embodiment, the mass-volume ratio of the mixture and water in the present invention is 1 g: 50 mL. As an embodiment, the temperature of the water in the present invention is 5-25 °C; as another embodiment, the temperature of the water in the present invention is 10-20 °C; as another embodiment, the temperature of the water in the present invention is 15 °C. As an embodiment, the soaking time in the present invention is 40-50 min; as another embodiment, the soaking time in the present invention is 45 min. The present invention soaks the mixture of Hovenia dulcis, Puerariae flos and Puerariae lobata, which has the advantages of promoting the dissolution of components and enhancing the curative effect.

[0041] After obtaining the soaked product, the present invention decocts the soaked product to obtain a decoction. As an embodiment, the decocting time in the present invention is ≥ 30 min; as another embodiment, the decocting time in the present invention is 120 min. The present invention decocts Hovenia dulcis, Puerariae flos and Puerariae lobata together. Compared with the method of decocting raw materials separately and then combining the decoction, it has the advantages of saving time cost and facilitating subsequent processing. Moreover, in the process of co-decocting Hovenia dulcis, Puerariae flos and Puerariae lobata, they synergistically enhance each other to enhance the effects of relieving alcohol toxicity, invigorating the spleen and regulating the stomach.

[0042] After obtaining the decoction, the present invention mixes the decoction and an organic acid at 80 °C to obtain a traditional Chinese medicine composition. As an embodiment, the mass-volume ratio of the organic acid and the decoction in the present invention is 20-40 g: 1 L; as another embodiment, the mass-volume ratio of the organic acid and the decoction in the present invention is 30 g: 1 L. As an embodiment, the mixing method at 80 °C in the present invention can be stirring at 80 °C. As an embodiment, the organic acid in the present invention includes one or more of citric acid, tartaric acid and malic acid; as another embodiment, the organic acid in the present invention is citric acid. The present invention mixes the organic acid and the decoction, which has the advantages of increasing drug dissolution and improving transdermal absorption.

[0043] The present invention also provides the application of the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition obtained by the preparation method described in the above technical solution in the preparation of products for relieving alcohol and protecting the liver and / or treating alcoholic liver disease.

[0044] As an embodiment, the product of the present invention includes a pharmaceutical preparation. As an embodiment, the dosage forms of the pharmaceutical preparation of the present invention include liposomes, pastes, and patches. The present invention uses the above-mentioned traditional Chinese medicine composition to prepare liposomes, pastes, and patches. Compared with common oral dosage forms, it can bypass the first-pass effect of the liver, reduce the decomposition of drugs during metabolism, increase the effective drug concentration, and further improve the effect of relieving hangover and protecting the liver.

[0045] The present invention also provides a liposome for relieving hangover and protecting the liver, which includes a traditional Chinese medicine composition and a thickening agent;

[0046] The traditional Chinese medicine composition is the traditional Chinese medicine composition described in the above technical solution or the traditional Chinese medicine composition obtained by the preparation method described in the above technical solution; the thickening agent includes carboxymethyl cellulose and / or gelatin.

[0047] As an embodiment, the mass-volume ratio of the thickening agent and the traditional Chinese medicine composition in the present invention is 5-50 g: 1 L; as another embodiment, the mass-volume ratio of the thickening agent and the traditional Chinese medicine composition in the present invention is 20 g: 1 L. As an embodiment, the thickening agent of the present invention includes carboxymethyl cellulose and gelatin, and the mass ratio of carboxymethyl cellulose and gelatin is 1:1-3:1.

[0048] The present invention also provides a preparation method of the liposome described in the above technical solution, which includes the following steps: dispersing the traditional Chinese medicine composition into an ethanol solution with a volume concentration of 30-80% by ultrasonic treatment to obtain a dispersion; mixing the dispersion and the thickening agent to obtain the liposome.

[0049] As an embodiment, the temperature of the ultrasonic treatment in the present invention is 10-50 °C; as another embodiment, the temperature of the ultrasonic treatment in the present invention is 25 °C. As an embodiment, the power of the ultrasonic treatment in the present invention is 50-500 W; as another embodiment, the power of the ultrasonic treatment in the present invention is 300 W. As an embodiment, the volume concentration of the ethanol solution in the present invention is 40-85%; as another embodiment, the volume concentration of the ethanol solution in the present invention is 75%. As an embodiment, the traditional Chinese medicine composition is dispersed into an ethanol solution with a volume concentration of 30-80% by ultrasonic treatment until the volume concentration of ethanol is 5%.

[0050] The present invention also provides a paste for relieving hangover and protecting the liver, which includes a liposome and a stabilizer;

[0051] The liposome is the liposome described in the above technical solution or the liposome obtained by the preparation method described in the above technical solution; the stabilizer includes a suspending agent and / or a surfactant.

[0052] As an implementation manner, the mass-volume ratio of the stabilizer to the liposome in the present invention is 2-20 g: 1 L; as another implementation manner, the mass-volume ratio of the stabilizer to the liposome in the present invention is 6 g: 1 L. As an implementation manner, the suspending agent in the present invention includes one or more of polyvinylpyrrolidone, gum arabic, xanthan gum, pectin, sodium alginate, propylene glycol alginate, microcrystalline cellulose, chitosan, dextran, carbomer. As an implementation manner, the surfactant in the present invention includes one or more of sodium dodecyl sulfate, polysorbate 80, polyethylene glycol, glyceryl stearate, alkylphenol polyoxyethylene ether, span, betaine, sucrose fatty acid ester, sodium lauroyl glutamate, octyl glucoside.

[0053] As an implementation manner, the paste in the present invention further includes tartaric acid. As an implementation manner, the mass-volume ratio of the tartaric acid to the liposome in the present invention is 0.5-2 g: 1 L; as another implementation manner, the mass-volume ratio of the tartaric acid to the liposome in the present invention is 1 g: 1 L. Adding tartaric acid to the paste in the present invention can further promote the dissolution of the traditional Chinese medicine composition, and has the effects of moisturizing and improving the performance of the paste.

[0054] The present invention also provides an anti-alcoholism and liver-protecting patch, which includes a breathable material and a paste coated on the breathable material; the paste is the paste in the above technical solution. As an implementation manner, the paste in the above technical solution is coated on the breathable material and then cut to obtain the patch. As an implementation manner, the paste in the above technical solution is made into a circle with a radius of 6 cm. The present invention has no strict requirements on the coating and cutting methods, and conventional operations can be used, such as using a coating and cutting machine.

[0055] In order to further illustrate the present invention, the following describes in detail a traditional Chinese medicine composition and liposome for anti-alcoholism and liver protection provided by the present invention, their preparation methods and applications with reference to the drawings and embodiments, but they cannot be understood as limiting the protection scope of the present invention.

[0056] All the data in the embodiments of the present invention were input into SPSS 22.0 software for analysis. If the measurement data conforms to the normal distribution, the One-Way ANOVA statistical method can be used, and p < 0.05 indicates that the difference is statistically significant.

[0057] Example 1

[0058] 1. Prepare materials according to the following mass parts: 10 parts of Hovenia dulcis, 10 parts of Pueraria lobata flower, and 10 parts of Pueraria lobata.

[0059] 2. Decoct: After mixing the raw materials in step 1, soak them in cold water for 45 min, with a material-water ratio of 1 g: 50 mL, and then decoct for 30 min after soaking.

[0060] 3. Preparation of Active Substance: Take a certain amount of citric acid and add it to the liquid obtained after decocting in Step 2. Stir at 80 °C until a gel is formed. Among them, the amount of citric acid is 30 g / L.

[0061] 4. Preparation of Liposome Composition: Add 75% v / v ethanol to the gel in Step 3 until the ethanol content is 5%. Ultrasonic at 25 °C until the gel is slightly dispersed in ethanol. After the gel is dispersed, add carboxymethyl cellulose; among them, the mass-volume ratio of carboxymethyl cellulose to the traditional Chinese medicine composition is 20 g: 1 L.

[0062] 5. Preparation of Paste: Add phosphatidylcholine to the liposome composition in Step 4 according to the mass-volume ratio of phosphatidylcholine to liposome of 6 g: 1 L; then, according to the mass-volume ratio of tartaric acid to liposome of 1 g: 1 L, add tartaric acid to ultrapure water. After mixing evenly, add it to the above liposome composition and stir to obtain a paste.

[0063] 6. Preparation of Patch: Transfer the prepared paste to a coating and cutting machine for uniform coating and cutting operations, and then place it in a cool environment for curing to obtain a gel transdermal patch carrying extracts of Hovenia dulcis, Puerariae flos, and Puerariae radix.

[0064] Example 2

[0065] Same as Example 1, the only difference is that the raw materials are prepared according to the following mass parts: 20 parts of Hovenia dulcis, 5 parts of Puerariae flos, and 8 parts of Puerariae radix.

[0066] Example 3

[0067] Same as Example 1, the only difference is that the raw materials are prepared according to the following mass parts: 5 parts of Hovenia dulcis, 8 parts of Puerariae flos, and 10 parts of Puerariae radix.

[0068] Example 4

[0069] Same as Example 1, the only difference is that the raw materials are prepared according to the following mass parts: 10 parts of Hovenia dulcis, 8 parts of Puerariae flos, and 5 parts of Puerariae radix.

[0070] Comparative Example 1

[0071] Same as Example 1, the only difference is that Hovenia dulcis is not used.

[0072] Comparative Example 2

[0073] Same as Example 1, the only difference is that Puerariae flos is not used.

[0074] Comparative Example 2

[0075] Same as Example 1, the only difference is that Puerariae radix is not used.

[0076] Test Example 1

[0077] 1. Experimental materials

[0078] 1.1 Experimental animals

[0079] Twenty-four SPF (Specific pathogen free) grade male C57BL / 6J mice, 6-8 weeks old, weighing 18-22 g, were purchased from Liaoning Changsheng Biotechnology Co., Ltd. and raised in the animal center of Liaoning University of Traditional Chinese Medicine. They had free access to water and were fed quantitatively. The bedding was changed three times a week, and the water bottles were changed once a week. The breeding environment was illuminated alternately with 12 hours of light and 12 hours of darkness, maintaining a temperature of 20-26 °C and a humidity of 40-70%. During the experimental operation process, the handling and intervention methods of the experimental mice strictly followed the regulations on the use and ethics of experimental animals of Liaoning University of Traditional Chinese Medicine.

[0080] 1.2 Experimental drugs

[0081] The gel transdermal patch obtained in Example 1.

[0082] 1.3 Main reagents

[0083] Liver function test kits were purchased from Nanjing Jiancheng Bioengineering Institute. 4% paraformaldehyde, gradient ethanol, xylene, hematoxylin stain, 1% eosin stain, neutral gum, Sirius red staining solution, and 0.5% acetic acid were purchased from Chuzhou Shinoda Biotechnology Co., Ltd.

[0084] 1.4 Main instruments and equipment

[0085] Paraffin slicer (Leica); automatic biochemical analyzer (Hitachi, Japan); microscope (Olympus); laminar flow hood (China); microplate reader (BIO-RAD).

[0086] 2. Experimental methods

[0087] 2.1 Animal grouping

[0088] Twenty-four male C57BL / 6J mice were randomly divided into four groups, A, B, C, and D (n = 6), namely group A (normal group), group B (liquid diet control group), group C (ethanol liquid diet group), and group D (ethanol liquid diet intervention group).

[0089] 2.2 Feed formula

[0090] Ordinary solid feed: crude protein ≥ 18%, crude fat ≥ 4%, crude fiber ≤ 5%, crude ash ≤ 8%. The calorific value of this ordinary solid feed is 3.4 Kcal / g;

[0091] Ordinary liquid feed: the proportion of each component is 35% fat, 18% protein, and 47% carbohydrate. The calorific value of this ordinary liquid feed is 1 kcal / mL;

[0092] Ethanol liquid diet: The proportions of each component are 35% fat, 18% protein, 19% carbohydrate, and 28% ethanol solution. In this ethanol liquid diet, the ethanol content is 5% V / V, and the calorie content is 2.8 kcal / mL.

[0093] 2.3 Model establishment and intervention methods

[0094] Group A (normal group): Fed according to the instructions of ordinary solid diet, with free access to food and water.

[0095] Group B (liquid diet control group): Fed according to the instructions of ordinary liquid diet, with water deprivation.

[0096] Group C (ethanol liquid diet group): According to the Gao-Binge model (Gao Bin, Chang Binxia, Xu Mingjiang. Mouse model of alcoholic liver disease with chronic ethanol feeding plus acute ethanol gavage (NIAAA model or Gao-Binge model). Infectious Disease Information. 2013. 26(05): 307-311.), fed according to the instructions of ethanol liquid diet, with water deprivation. At 7-9 am on the 16th day, gavaged with 31.5% (v / v) ethanol solution, and animal specimens were taken 9 h later.

[0097] Group D (ethanol liquid diet intervention group): The diet feeding was the same as that of Group C, and at the same time, the gel transdermal patch obtained in Example 1 was applied at the sites where the thoracic sympathetic nerves of the spine at T7-T9 and the liver area emerged.

[0098] 2.4 Sample collection

[0099] In Experiment 1, after 12 weeks, all mice were fasted and water-deprived for 12 h, then anesthetized, weighed, bled, and serum was separated. The liver tissue was separated and weighed, and a 1 cm 3 sized sample was cut from the fixed part of the right liver lobe and immersed in 4% paraformaldehyde for fixation for pathological section preparation. The remaining liver parts were stored in a -80 °C ultra-low temperature refrigerator for later use.

[0100] 2.5 Observation indicators and measurement methods

[0101] 2.5.1 General condition of mice

[0102] Observe the hair color luster, behavior performance, mental state, nutritional status of mice in each group, record the change of mouse body weight every 2 days, and record carefully.

[0103] The results showed that no mice died during the whole experiment. Mice in Group C showed irritable and excitable states, with greasy and knotted fur; mice in Group A and Group B had no obvious abnormal behaviors, with bright fur; compared with the model group (Group C), the general conditions of mice in Group D were improved to varying degrees.

[0104] 2.5.2 Mouse body weight and liver coefficient

[0105] After blood collection, the mice were fixed, the abdominal cavity was opened, the liver was taken out and weighed. The liver coefficient = liver (g) / body weight (g). The results are shown in Table 1.

[0106] Table 1 Comparison results of body weight and liver / body weight of mice in each group

[0107] Group Body weight (g) Liver coefficient (%) A 20.5±1.33 1.40±0.08 B 21.2±0.67 1.32±0.10 C <![CDATA[25.79±1.03 *※ > <![CDATA[1.05±0.07 *※ > D <![CDATA[23.04.1±0.99 #※ > <![CDATA[1.35±0.06 # >

[0108] Note: Compared with group A, * p < 0.05; compared with group B, ※ p < 0.05; compared with group C, # p < 0.05, the same below.

[0109] At the beginning and end of the experiment, the body weight baselines of the mice in each group were consistent. According to Table 1, it can be seen that compared with group A and group B, the body weight and liver index of group C were significantly increased (p < 0.05); compared with group C, the liver index of group D was significantly decreased (p < 0.05); compared with group B, the body weight of group D was significantly increased (p < 0.05).

[0110] 2.5.3 Detection of serological indicators

[0111] The mice were bled by eye enucleation. After sufficient shaking before centrifugation, the serum was separated at a speed of 3000 revolutions per minute (rpm). The supernatant was taken and the precipitate was discarded into a new centrifuge tube and stored frozen at -80 °C to avoid repeated freezing and thawing. The specific operations for detecting various indicators of liver function and blood lipids were carried out according to the instructions of the corresponding reagent kits. The results are shown in Table 2.

[0112] Table 2 Comparison results of liver function and blood lipids of mice in each group

[0113] Group ALT AST TG CHOL A 165.2±30.5 43.1±59.1 0.81±0.11 3.0±0.5 B 200.9±41.2 50.9±42.1 0.79±0.14 3.8±0.7 C <![CDATA[554.1±69.1* ※ > <![CDATA[213.1±60.4* ※ > <![CDATA[1.03±0.32* ※ > <![CDATA[6.6±0.9* ※ > D <![CDATA[420.5±53.2* ※# > <![CDATA[100.3±39.9* ※# > <![CDATA[0.99±0.49* ※ > <![CDATA[4.9±1.1* ※ >

[0114] According to Table 2, it can be seen that compared with group A and group B, the liver function (ALT, AST) and blood lipid levels (TG, CHOL) of the mice in group C were significantly increased (p < 0.05); compared with group C, the liver function of the mice in group D was improved to varying degrees (p < 0.05), and the blood lipid improvement was not obvious.

[0115] 2.5.4 Liver pathological examination

[0116] 2.5.4.1 Preparation of pathological sections

[0117] The mouse liver was fixed with 4% paraformaldehyde for 24 hours. The liver tissue was trimmed flat and subjected to gradient ethanol dehydration treatment, placed in 75% ethanol for 240 min, 85% ethanol for 120 min, 90% ethanol for 120 min, 95% ethanol for 60 min, and absolute ethanol for a total of 60 min. Clearing: Immersed in xylene, 30 minutes each time, for a total of 3 times. After clearing, the liver was cut into pieces and placed in an embedding machine to melt paraffin at 65 °C, and the liver pathology was immersed 3 times, 1 hour each time. The liver tissue was placed flat in the embedding cassette with the cut surface facing down for embedding. Sectioning: The thickness of each tissue section was set to 4 - 5 μm. After sectioning, it was placed flat in warm water, and the tissue section was gently lifted with a glass slide and dried in an oven at 60 °C for 2 h for standby. Deparaffinization: 2 times with xylene, 2 times with absolute ethanol, 1 time with 95% ethanol, 1 time with 90% ethanol, 1 time with 85% ethanol, 1 time with 75% ethanol, and 1 time with distilled water, 10 minutes each time.

[0118] 2.5.4.2 HE staining

[0119] Stained with hematoxylin for 3 min, and the floating color was washed off with distilled water. Differentiating solution (1% hydrochloric acid ethanol) was added dropwise for differentiation, and soaked in distilled water for 15 min. After bluing with 1% ammonia water, the excess water was washed off. Eosin was added dropwise and stained for 10 min, and rinsed with distilled water. Dehydration and clearing: 2 times with 95% ethanol, 1 min each time; 2 times with absolute ethanol, 5 min each time; 3 times with xylene, 5 min each time. Sealing the slide: A drop of neutral gum was added dropwise for sealing, and sealed with a microscope cover glass. The staining results are as Figure 1 shown.

[0120] According to Figure 1 It can be seen that under high magnification, the hepatic plates in the liver tissues of mice in group A and group B were normally distributed, and the hepatocytes were regularly arranged around the central vein of the liver. The hepatic nuclei and cell membranes in the hepatic plates were intact and clear, and the nucleoli were centered. In the HE staining of group C mice, the hepatic plate structure was disordered and unclear, a large amount of lipid droplets and fat vacuoles were accumulated in the hepatocytes, the distribution of hepatic nuclei shifted to the left or right, and a large number of inflammatory cells were infiltrated in the cytoplasm. A large amount of fibrous tissue hyperplasia was seen in Sirius red staining. In group D mice, the morphology and structure of the liver tissue were improved to varying degrees, the fat vacuoles and lipid droplets were significantly reduced compared with the model group, and only a few individual lipid droplets were present. The size of hepatocytes was normal, the morphology was relatively regular, the structure was relatively clear, and no inflammatory cells were seen.

[0121] 2.5.4.3 Sirius red staining

[0122] Add hematoxylin staining solution dropwise for 20 minutes, and rinse with distilled water for 30 s to wash away the excess staining solution. Add 1% hydrochloric acid ethanol for differentiation, and wash with distilled water for 10 min. Sirius red staining solution for 1 h, and rinse with distilled water for 30 s. Dehydration and clearing: absolute ethanol for 3 times, 1 min each time; xylene for 2 times, 1 min each time. Sealing the slide: Drop a drop of neutral balsam for sealing, cover the slide with a microscope cover glass, and the staining result is as Figure 2 shown.

[0123] According to Figure 2 it can be seen that there is a large amount of fibrotic deposition near the central vein of the liver tissue in group C, and the fibrotic tissue deposition in the liver tissue of group D has been improved to varying degrees.

[0124] According to the above content, it can be seen that the technical solution provided by the present invention has a definite effect on improving liver function and liver pathology caused by ethanol in mice, and has no obvious effect on the regulation of blood lipids.

[0125] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on this embodiment without creative efforts, and these embodiments all belong to the protection scope of the present invention.

Claims

1. A traditional Chinese medicine composition for relieving hangover and protecting the liver, characterized in that, It comprises raw materials in the following parts by mass: 1-100 parts of Hovenia dulcis, 1-100 parts of Puerariae flos, and 1-100 parts of Puerariae radix.

2. The traditional Chinese medicine composition according to claim 1, wherein The dosage form of the traditional Chinese medicine composition includes a gel.

3. The preparation method of the traditional Chinese medicine composition according to claim 1 or 2, characterized in that, It includes the following steps: Soak the mixture of Hovenia dulcis, Puerariae flos, and Puerariae radix with water to obtain the soaked product; the mass-volume ratio of the mixture to water is 1 g: 25-60 mL, the temperature of the water is 5-25 °C; the soaking time is 30-60 min; Decoct the soaked product to obtain a decoction. Mix the decoction and an organic acid at 80 °C to obtain the traditional Chinese medicine composition.

4. The preparation method according to claim 3, characterized in that, The decocting time ≥ 30 min; The mass-volume ratio of the organic acid to the decoction is 20-40 g: 1 L.

5. Use of the traditional Chinese medicine composition according to claim 1 or 2 or the traditional Chinese medicine composition obtained by the preparation method according to claim 3 or 4 in the preparation of a product for relieving alcohol and protecting the liver and / or treating alcoholic liver disease.

6. The application according to claim 5, wherein The product includes a pharmaceutical preparation; the dosage form of the pharmaceutical preparation includes liposomes, ointments, and patches.

7. A liposome for relieving hangover and protecting the liver, characterized in that, It includes a traditional Chinese medicine composition and a thickening agent; The traditional Chinese medicine composition is the traditional Chinese medicine composition according to claim 1 or 2 or the traditional Chinese medicine composition obtained by the preparation method according to claim 3 or 4; The thickening agent includes carboxymethyl cellulose and / or gelatin.

8. The method for preparing the liposome according to claim 7, characterized in that, It includes the following steps: disperse the traditional Chinese medicine composition into an ethanol solution with a volume concentration of 30-80% by ultrasonic treatment to obtain a dispersion; mix the dispersion and the thickening agent to obtain the liposomes.

9. A paste for relieving hangover and protecting the liver, characterized in that, It includes liposomes and a stabilizer; The liposomes are the liposomes according to claim 7 or the liposomes obtained by the preparation method according to claim 8; The stabilizer includes a suspending agent and / or a surfactant.

10. A patch for relieving hangover and protecting the liver, characterized in that, It includes a breathable material and an ointment coated on the breathable material; the ointment is the ointment according to claim 9.

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