A traditional Chinese medicine composition for preventing and treating fatty liver, a preparation method thereof and application thereof

Abstract

This invention relates to the field of traditional Chinese medicine (TCM) technology, and particularly to a TCM composition for the prevention and treatment of fatty liver, its preparation method, and its application. The TCM composition is made from raw hawthorn, dried tangerine peel, cassia seed, salvia miltiorrhiza, jujube seed, and raw licorice. Experiments show that this TCM composition can significantly reverse the increase in body weight and liver tissue weight in mice with non-alcoholic fatty liver disease, reverse the increase in serum TC, TG, LDL-C, AST, and ALT, as well as the increase in TC and TG in the liver, improve the pathological state of the liver, and protect the liver. The effect is significantly higher than that of any single component in the formula, or the absence or substitution of the formula, indicating that each herb has a synergistic effect. Furthermore, the TCM composition provided by this invention is simple to prepare, low in cost, and all herbs are medicinal and edible herbs, ensuring good safety.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to a traditional Chinese medicine composition for the prevention and treatment of fatty liver, its preparation method, and its application. Background Technology

[0002] Non-alcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized by excessive fat deposition in hepatocytes, excluding those caused by alcohol or other clearly defined liver-damaging factors. NAFLD is a common liver disease and is associated with a variety of other conditions, including non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, cirrhosis, and hepatocellular carcinoma. With the global prevalence of obesity and related metabolic syndromes, NAFLD has become a significant cause of chronic liver disease in developed countries such as Europe and the United States, as well as in affluent regions of my country. The prevalence of NAFLD in the general adult population is 10%–30%, of which 10%–20% are non-alcoholic steatohepatitis, and the incidence of cirrhosis within 10 years is as high as 25%, seriously endangering human health.

[0003] Currently, no regulatory agency has approved any drugs for the treatment of non-alcoholic fatty liver disease (NAFLD). The only internationally recommended treatments are pioglitazone and vitamin E, but these have certain adverse effects. Therefore, the discovery of new treatments for NAFLD remains necessary. Traditional Chinese medicine believes that the pathogenesis of fatty liver is rooted in qi stagnation and blood stasis, with damp-heat in the liver and gallbladder as the superficial manifestation. Treatment should focus on clearing heat and soothing the liver, promoting blood circulation and removing blood stasis, drying dampness, strengthening the spleen, and resolving phlegm.

[0004] With the fast pace of modern life and intensified social competition, many people overwork themselves due to pressure from work, study, and social engagements, severely impacting their health. Non-alcoholic fatty liver disease is becoming a real health problem troubling many. Existing traditional Chinese medicine (TCM) compositions have limitations in efficacy; therefore, it is necessary to provide a safe, effective, and definitively effective TCM composition. Summary of the Invention

[0005] In view of this, the present invention provides a traditional Chinese medicine composition for the prevention and treatment of fatty liver, its preparation method and application.

[0006] In view of this, the present invention provides a traditional Chinese medicine composition for the prevention and treatment of non-alcoholic fatty liver disease, its preparation method and application.

[0007] The traditional Chinese medicine composition provided by this invention includes six ingredients: raw hawthorn, dried tangerine peel, cassia seed, salvia miltiorrhiza, jujube seed, and raw licorice.

[0008] Furthermore, the traditional Chinese medicine composition provided by the present invention, by weight, comprises the following raw materials: 15-25 parts of raw hawthorn, 10-20 parts of dried tangerine peel, 10-15 parts of cassia seed, 8-12 parts of salvia miltiorrhiza, 10-20 parts of jujube seed, and 10-20 parts of raw licorice.

[0009] This invention has discovered that a traditional Chinese medicine composition obtained by combining the above-mentioned six medicinal and edible ingredients in a specific ratio can significantly improve non-alcoholic fatty liver disease induced by a high-fat diet. The formula is based on the principles of "clearing heat and soothing the liver, promoting blood circulation and removing blood stasis, drying dampness, strengthening the spleen, and resolving phlegm." Hawthorn is the principal ingredient, promoting digestion, resolving food stagnation, and dispersing blood stasis; tangerine peel regulates qi, strengthens the spleen, dries dampness, and resolves phlegm, serving as the assistant ingredients; cassia seed clears heat, improves eyesight, and moistens the intestines to relieve constipation; salvia miltiorrhiza promotes blood circulation and removes blood stasis; and jujube seed nourishes the heart and liver, serving as the adjuvant ingredients to help clear liver heat, resolve blood stasis, and calm the mind; licorice root harmonizes the other ingredients, serving as the guiding ingredient. The combined effects of these ingredients are to soothe the liver and strengthen the spleen, promote blood circulation, remove blood stasis, and resolve dampness and phlegm, targeting the pathogenesis of qi stagnation, blood stasis, and phlegm dampness in non-alcoholic fatty liver disease. Furthermore, all ingredients in the formula are medicinal and edible, ensuring high safety.

[0010] In some specific embodiments, the traditional Chinese medicine composition of the present invention is prepared from the following raw materials in parts by weight: 18 parts of raw hawthorn, 15 parts of dried tangerine peel, 13 parts of cassia seed, 10 parts of salvia miltiorrhiza, 12 parts of jujube seed, and 15 parts of raw licorice.

[0011] In some specific embodiments, the traditional Chinese medicine composition of the present invention is prepared from the following raw materials in parts by weight: 18 parts of raw hawthorn, 12 parts of dried tangerine peel, 13 parts of cassia seed, 10 parts of salvia miltiorrhiza, 10 parts of jujube seed, and 10 parts of raw licorice.

[0012] In some specific embodiments, the traditional Chinese medicine composition of the present invention is prepared from the following raw materials in parts by weight: 18 parts of raw hawthorn, 18 parts of dried tangerine peel, 13 parts of cassia seed, 10 parts of salvia miltiorrhiza, 20 parts of jujube seed, and 20 parts of raw licorice.

[0013] The present invention also provides a method for preparing the traditional Chinese medicine composition, comprising: mixing raw hawthorn, dried tangerine peel, cassia seed, salvia miltiorrhiza, jujube seed and raw licorice, extracting with solvent, collecting the extract and concentrating it.

[0014] In some implementations, the solvent is water or an aqueous solution of ethanol.

[0015] In some embodiments, the volume fraction of the ethanol-water solution is 10%-95%, or any value within this concentration range, such as 10%, 20%, 50%, 70%, 90%, etc., preferably 70%.

[0016] In some embodiments, the extraction includes reflux extraction; the extraction can be performed once or multiple times, such as 1 to 3 times, specifically 1, 2, or 3 times. The extraction is carried out according to the following ratio: 5-20 mL of solvent is added for every 1 g of total raw material; when the total raw material is 100 g, 500 mL to 2000 mL of solution is added, specifically 500 mL, 1000 mL, 1200 mL, 1500 mL, 1800 mL, or 2000 mL of solvent can be added, preferably 1000 mL of solvent.

[0017] In some implementations, the drying process involves vacuum freeze drying or conventional drying followed by pulverization.

[0018] The present invention also provides a traditional Chinese medicine extract, which is obtained by extraction from the traditional Chinese medicine composition described in the present invention or by the preparation method described in the present invention.

[0019] The present invention also provides the application of the traditional Chinese medicine composition, the traditional Chinese medicine composition prepared by the preparation method, or the traditional Chinese medicine extract in the preparation of a drug for the prevention and treatment of non-alcoholic fatty liver disease.

[0020] In some implementations, the prevention and treatment of non-alcoholic fatty liver disease includes: reducing body weight and / or liver weight; and / or reducing the level of at least one of the following in serum: alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), total cholesterol (TC), and low-density lipoprotein (LDL); and / or reducing the content of triglycerides (TG) and / or total cholesterol (TC) in liver tissue; and / or improving the pathological state of hepatic steatosis.

[0021] Experiments show that the herbal composition of this invention can significantly reverse the increase in body weight and liver weight in mice with non-alcoholic fatty liver disease induced by high-fat diet, significantly reduce serum ALT, AST, TG, TC and LDL levels, and significantly reduce TG and TC content in the liver, and improve pathological conditions such as hepatic steatosis and fat vacuoles. Its effect is significantly better than any single herb in the formula, missing ingredients, and substitute ingredients, indicating that the combination of each herb is reasonable and synergistic.

[0022] The present invention also provides a pharmaceutical composition for the prevention and treatment of non-alcoholic fatty liver disease, comprising the traditional Chinese medicine composition described in the present invention, the traditional Chinese medicine composition prepared by the preparation method described in the present invention, or the traditional Chinese medicine extract described in the present invention, as well as pharmaceutically acceptable excipients.

[0023] In the pharmaceutical composition of the present invention, the pharmaceutically acceptable excipients include at least one of fillers, disintegrants, lubricants, flavoring agents, binders, and coating materials.

[0024] The dosage form of the drug described in this invention can be any pharmaceutically acceptable dosage form, including but not limited to oral formulations, injectable formulations, transdermal absorption formulations, etc.

[0025] In some implementations, the dosage form of the drug is an oral dosage form, which includes, but is not limited to, tablets, capsules, granules, pills, powders, syrups, oral solutions, oral suspensions, etc.

[0026] In some implementations, the dosage form of the drug is an injectable preparation, including but not limited to pharmaceutically acceptable injectable dosage forms such as injection solutions, injection suspensions, injection emulsions, and injection powders.

[0027] In some implementations, the dosage of the drug is a transdermal absorption formulation.

[0028] The traditional Chinese medicine composition provided by this invention is made from raw hawthorn, dried tangerine peel, cassia seed, salvia miltiorrhiza, jujube seed, and raw licorice, all of which are medicinal and edible herbs with high safety.

[0029] Compared with the prior art, the present invention has the following beneficial technical effects:

[0030] 1. The herbal composition of this invention is a compound made from the medicinal and edible herbs hawthorn, tangerine peel, cassia seed, salvia miltiorrhiza, jujube seed, and licorice root. It is used to improve non-alcoholic fatty liver disease. Hawthorn is the principal herb, which promotes digestion, invigorates blood circulation, removes blood stasis, lowers lipids, and eliminates turbidity. Salvia miltiorrhiza invigorates blood circulation, removes blood stasis, and improves liver microcirculation. Cassia seed moistens the intestines, promotes bowel movement, lowers lipids, and improves eyesight, serving as the assistant herb. Tangerine peel regulates qi, strengthens the spleen, dries dampness, and resolves phlegm. Jujube seed nourishes the liver and calms the mind, acting as the adjuvant herb. Licorice root tonifies the spleen, replenishes qi, and harmonizes the other herbs. The six herbs work together to soothe the liver, strengthen the spleen, invigorate blood circulation, lower lipids, and improve non-alcoholic fatty liver disease.

[0031] 2. The combination of herbs in this invention has the effects of soothing the liver and strengthening the spleen, promoting blood circulation and lowering lipids, and improving non-alcoholic fatty liver disease. Systematic pharmacological studies have shown that this traditional Chinese medicine composition has the effect of improving non-alcoholic fatty liver disease, and the combination of individual herbs in the formula has the best synergistic effect, which is significantly better than that of individual herbs or alternative formulas.

[0032] 3. The traditional Chinese medicine composition of the present invention not only has a therapeutic effect on non-alcoholic fatty liver disease, but also has good safety due to its food-medicine homology composition. It has the potential value of shifting the health protection focus forward, managing chronic diseases and promoting national health. Attached Figure Description

[0033] Figure 1 HE staining results for mouse liver (scale: 50μm);

[0034] Figure 2 Oil Red O staining results for mouse liver (ruler: 50 μm). Detailed Implementation

[0035] This invention provides a traditional Chinese medicine composition for the prevention and treatment of fatty liver, its preparation method, and its application. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired result. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The method and application of this invention have been described through preferred embodiments. Those skilled in the art can obviously make modifications or appropriate alterations and combinations to the method and application described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0036] In this document, the terms "including", "comprising", and "having" describe both closed-loop technical solutions consisting of the listed features and open-loop technical solutions that include the listed features.

[0037] In this document, the term “and / or” as used includes any and all combinations of one or more of the related listed items.

[0038] In this document, numerical ranges are referred to as continuous unless otherwise specified, and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when a range refers to an integer, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0039] In this article, when referring to units of data ranges, if the unit is only followed by the right endpoint, it means that the units of the left and right endpoints are the same. For example, "15~25 parts" means that the units of the left endpoint "15" and the right endpoint "25" are both "parts".

[0040] The test materials used in this invention are all common commercial products and can be purchased on the market.

[0041] The present invention will be further illustrated below with reference to the embodiments:

[0042] This invention provides a traditional Chinese medicine composition with efficacy in improving non-alcoholic fatty liver disease and its preparation method. Those skilled in the art can refer to the content of this document and appropriately modify the process parameters to achieve the desired result. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art can obviously make modifications or appropriate alterations and combinations to the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0043] This invention provides a traditional Chinese medicine composition and its preparation method that improve non-alcoholic fatty liver disease (NAFLD). Through in-depth research and creative work, the inventors have discovered a traditional Chinese medicine composition and its preparation method that demonstrates clear efficacy and enhanced safety in treating NAFLD. The complete formula of this invention exhibits a significant synergistic effect in the prevention and treatment of NAFLD, significantly improving related symptoms and pathological changes. Its preventive and therapeutic effects are significantly superior to extracts of single herbs or extracts from combinations of single herbs. This traditional Chinese medicine composition is suitable for long-term use, meeting clinical needs for the prevention and treatment of NAFLD and possessing promising application prospects.

[0044] The numerical ranges and parameters involved in this invention have been presented as precisely as possible in the specific embodiments. However, any numerical value inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise expressly stated, it should be understood that all numerical ranges or specific data used in this disclosure may have a reasonable deviation within a certain range, such as ±10%, ±5%, ±1%, or ±0.5%.

[0045] The embodiments and comparative examples of this invention describe some cases. These embodiments illustrate certain implementations of the invention. However, this does not mean that the effects of the invention can only be achieved in these cases. In fact, any concentration of each component between the two endpoints shown in the embodiments (especially Examples 1-3) can achieve a good effect in improving non-alcoholic fatty liver disease, with Example 1 showing the best effect. The comparative examples only list some cases where the effects were not good in the experiments. In addition, many other attempts were made during the research and development process, such as using different medicinal ingredients or different dosages, but the effects of these attempts were not as good as those of Examples 1-3, and will not be elaborated here.

[0046] All test materials used in this invention are commercially available products. The Chinese herbal raw materials used in the experiments of this invention, including raw hawthorn, dried tangerine peel, cassia seed, salvia miltiorrhiza, jujube seed, and raw licorice, were all purchased from Beijing Tongrentang (Group) Co., Ltd. The invention is further illustrated below with reference to the embodiments.

[0047] Example 1: Preparation of Traditional Chinese Medicine Extracts

[0048] 1. Preparation of the extract of composition 1 (complete formula)

[0049] Take 18g of raw hawthorn, 15g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, 12g of jujube seed, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time heating under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 1.

[0050] 2. Preparation of extracts from combination 2 (different proportions of the whole formula)

[0051] Take 18g of raw hawthorn, 12g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, 10g of jujube seed, and 10g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time heating under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 2.

[0052] 3. Preparation of extracts from composition 3 (different proportions of the whole formula)

[0053] Take 18g of raw hawthorn, 18g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, 20g of jujube seed, and 20g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time heating under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 3.

[0054] 4. Preparation of the extract of composition 4 (raw hawthorn + dried tangerine peel + cassia seed + salvia miltiorrhiza + jujube seed)

[0055] Take 18g of raw hawthorn, 15g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, and 12g of jujube seed; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 4.

[0056] 5. Preparation of the extract of composition 5 (raw hawthorn + dried tangerine peel + cassia seed + salvia miltiorrhiza + raw licorice)

[0057] Take 18g of raw hawthorn, 15g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time heating under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 5g.

[0058] 6. Preparation of the extract of composition 6 (raw hawthorn + dried tangerine peel + cassia seed + jujube seed + raw licorice)

[0059] Take 18g of raw wheat, 15g of dried tangerine peel, 13g of cassia seed, 12g of jujube seed, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 6.

[0060] 7. Preparation of the extract of composition 7 (raw hawthorn + dried tangerine peel + salvia miltiorrhiza + jujube seed + raw licorice)

[0061] Take 18g of raw hawthorn, 15g of dried tangerine peel, 10g of salvia miltiorrhiza, 12g of jujube seed, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time heating under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 7.

[0062] 8. Preparation of extract from composition 8 (raw hawthorn + cassia seed + salvia miltiorrhiza + jujube seed + raw licorice)

[0063] Take 18g of raw hawthorn, 13g of cassia seed, 10g of salvia miltiorrhiza, 12g of jujube seed, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time heating under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 8.

[0064] 9. Preparation of the extract of composition 9 (tangerine peel + cassia seed + salvia miltiorrhiza + jujube seed + licorice root)

[0065] Take 15g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, 12g of jujube seed, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time under reflux for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 9.

[0066] 10. Preparation of raw hawthorn extract

[0067] 18g of raw hawthorn was soaked in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then refluxed for 2 hours. The mixture was filtered, and the filtrate was collected. The residue was extracted twice with distilled water under the same conditions, each time refluxed for 2 hours. The extracts were filtered, and the filtrates were collected. The three filtrates were combined, concentrated under reduced pressure, and freeze-dried under vacuum to obtain 10g of extract.

[0068] 11. Preparation of Tangerine Peel Extract

[0069] Soak 15g of dried tangerine peel at room temperature for 12 hours, then reflux for 2 hours. Filter and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time refluxed for 2 hours. Filter the extracts and collect the filtrates. Combine the three filtrates, concentrate under reduced pressure, and freeze-dry under vacuum to obtain extract 11.

[0070] 12. Preparation of Cassia Seed Extract

[0071] 13g of cassia seeds were soaked in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then refluxed for 2 hours. The mixture was filtered, and the filtrate was collected. The residue was extracted twice with distilled water under the same conditions, each time refluxed for 2 hours. The extracts were filtered, and the filtrates were collected. The three filtrates were combined, concentrated under reduced pressure, and freeze-dried under vacuum to obtain extract 12.

[0072] 13. Preparation of Salvia miltiorrhiza extract

[0073] 10g of Salvia miltiorrhiza was soaked in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then refluxed for 2 hours. The mixture was filtered, and the filtrate was collected. The residue was extracted twice with distilled water under the same conditions, each time refluxed for 2 hours. The extracts were filtered, and the filtrates were collected. The three filtrates were combined, concentrated under reduced pressure, and freeze-dried under vacuum to obtain extract 13.

[0074] 14. Preparation of Ziziphus jujuba seed extract

[0075] 12g of jujube seed was soaked in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then refluxed for 2 hours. The mixture was filtered, and the filtrate was collected. The residue was extracted twice with distilled water under the same conditions, each time refluxed for 2 hours. The extracts were filtered, and the filtrates were collected. The three filtrates were combined, concentrated under reduced pressure, and freeze-dried under vacuum to obtain extract 14.

[0076] 15. Preparation of raw licorice extract

[0077] 15g of raw licorice root was soaked in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then refluxed for 2 hours. The mixture was filtered, and the filtrate was collected. The residue was extracted twice with distilled water under the same conditions, each time refluxed for 2 hours. The extracts were filtered, and the filtrates were collected. The three filtrates were combined, concentrated under reduced pressure, and freeze-dried under vacuum to obtain extract 15.

[0078] Example 2: Preparation of granules of traditional Chinese medicine composition (composition 1)

[0079] Formula: 18g raw hawthorn, 15g dried tangerine peel, 13g cassia seed, 10g salvia miltiorrhiza, 12g jujube seed, and 15g raw licorice root;

[0080] Take 18g of raw hawthorn, 15g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, 12g of jujube seed, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time refluxed for 2 hours. Filter the extract and collect the filtrate. Combine the three filtrates, concentrate under reduced pressure to a relative density of 1.10–1.20 (25℃), dry, pulverize into a fine powder, add dextrin at a ratio of 1:1–1:3, mix evenly, granulate, dry, and granulate to 10–20 mesh. Fill into double-sided aluminum bags.

[0081] Example 3: Preparation of tablets of traditional Chinese medicine composition (composition 1)

[0082] Formula: 18g raw hawthorn, 15g dried tangerine peel, 13g cassia seed, 10g salvia miltiorrhiza, 12g jujube seed, and 15g raw licorice root;

[0083] Take 18g of raw hawthorn, 15g of dried tangerine peel, 13g of cassia seed, 10g of salvia miltiorrhiza, 12g of jujube seed, and 15g of raw licorice root; soak them in 70% ethanol (1:10, m:V) at room temperature for 12 hours, then heat under reflux for 2 hours, filter, and collect the filtrate. Extract the residue twice with distilled water under the same conditions, each time under reflux for 2 hours. Filter the extract and collect the filtrate. Combine the three filtrates, concentrate under reduced pressure to a relative density of 1.10–1.20 (25℃), dry, and pulverize into a fine powder. Place the above raw and excipient mixture into a one-step granulator, add starch, lactose, and microcrystalline cellulose (1:1:2), granulate, and dry. After determining the moisture content to be <2%, granulate through a 10–20 mesh sieve, add 0.1–1% micronized silica gel and magnesium stearate, and mix well. Compress using an 8#–9# die, controlling the tablet pressure at 20–40 Newtons. The compressed tablets are placed in a coating machine and wrapped in a moisture-proof and light-proof coating, with the weight gain controlled at 2%–4%.

[0084] Example 1: Comparative evaluation of the effects of different traditional Chinese medicine combinations on improving non-alcoholic fatty liver disease

[0085] (a) Laboratory animals

[0086] ICR mice, male, 18-22g, were purchased from Changchun Yisi Experimental Animal Technology Co., Ltd.

[0087] (ii) Test Sample

[0088] The test samples in experimental groups 1-15 were extracts 1-15 prepared in Example 1. Based on the human clinical dose, the mouse dose was converted according to the body surface area method. The mouse administration dose was 10.79g crude drug / kg (mouse body weight). Administration method: extracts 1-15 were added to distilled water to make solutions for gavage.

[0089] The comparative example test sample was prepared according to the "Traditional Chinese Medicine Composition (I)" disclosed in patent application number 202411131273.X, which consists of: 15 parts of Salvia miltiorrhiza, 15 parts of Cassia tora, 15 parts of raw hawthorn, 6 parts of raw licorice, 15 parts of raw Astragalus membranaceus, and 6 parts of dried tangerine peel. The comparative example test sample was obtained by preparing extract 1 according to the method described in this application. The dosage for mice was 10.79 g of crude drug / kg (mouse body weight).

[0090] (III) Experimental Instruments

[0091] Electronic balance: Shanghai Sunny Hengping Scientific Instruments Co., Ltd. Dehydrator: DIAPATH. Embedding machine: Wuhan Junjie Electronics Co., Ltd. Pathology slide machine: Shanghai Leica Instruments Co., Ltd. Tissue spreader: Zhejiang Jinhua Kedi Instrument Equipment Co., Ltd. Microscope: OLYMPUS JAPAN

[0092] (iv) Grouping and administration

[0093] Six-week-old male C57BL / 6J mice were used in the experiment to induce a non-alcoholic fatty liver disease (NAFLD) model using a high-fat diet induced by HF60. After one week of acclimatization, the C57BL / 6J mice were randomly divided into 19 groups based on their body weight: a normal control group, a model group, a positive control group (atorvastatin, 5 mg / kg), experimental groups 1-15 (administered extracts 1-15 respectively), and a control group, with eight mice in each group. The normal control group was fed a normal diet daily, while the other groups were fed a high-fat diet to establish the NAFLD model for six consecutive weeks. After six weeks, the mice were administered the drug via gavage at 8:30 AM and 3:30 PM daily at a dose of 0.2 mL / 10 g for six consecutive weeks. The normal control group and the model group received saline solution based on body weight, experimental groups 1-15 received the corresponding experimental drug solution based on body weight, the control group received the control drug solution, and the positive control group received atorvastatin.

[0094] (V) Indicator Testing:

[0095] 1. Mouse body weight determination:

[0096] After 12 weeks, the body weight of all mice was measured and recorded. The results are shown in Table 1.

[0097] Table 1 Effects of each drug group on mouse body weight

[0098]

[0099] Note: # indicates comparison with the blank control group, #p<0.05, ##p<0.01; * indicates comparison with the model group, *p<0.05, **p<0.01.

[0100] The experimental results in Table 1 show that compared with the blank control group, the body weight of mice in the model group was significantly increased (p<0.01). Compared with the model group, the body weight of mice in experimental groups 1-9 was significantly decreased. The differences in experimental groups 1-3 were highly significant (p<0.01), and the differences in experimental groups 4-9 were significant (p<0.05). The body weight of mice in experimental groups 10-15 decreased, but the difference was not statistically significant. This indicates that compositions 1-9 can significantly reduce the increase in body weight in mice with non-alcoholic fatty liver disease (NAFLD), with compositions 1-3 showing significantly better effects than compositions 4-9, and composition 1 showing the best effect. This suggests that the components of the traditional Chinese medicine in composition 1 have a significant synergistic effect and the optimal ratio, making its effect in reducing the increase in body weight in NAFLD mice stronger than the individual application of any component in the formula. Compared with the control group, compositions 1-3 showed a stronger effect in reducing body weight than the traditional Chinese medicine compositions in the control group.

[0101] 2. Liver weight and liver coefficient:

[0102] After blood was collected by enucleation of the eyeballs, the mice were euthanized, and the livers were quickly removed and washed with pre-cooled physiological saline. The moisture was blotted dry with filter paper, and the livers were weighed on a balance. The results are shown in Table 2.

[0103] Table 2. Liver weight measurement results in mice of each drug group

[0104]

[0105] Note: # indicates comparison with the blank control group, #p<0.05, ##p<0.01; * indicates comparison with the model group, *p<0.05, **p<0.01.

[0106] The experimental results in Table 2 show that, compared with the blank control group, the liver weight of mice in the model group was significantly increased (p<0.01). Compared with the model group, the liver weight of mice in experimental groups 1-15 was significantly decreased (p<0.01, p<0.05), but the decrease in experimental group 1 was the most significant. This suggests that the traditional Chinese medicine in composition 1 has a synergistic effect and the compatibility ratio is optimal. Compared with the comparative group, compositions 1-3 showed a stronger improvement in liver weight than the traditional Chinese medicine composition in the comparative group.

[0107] 3. Determination of four indicators of liver function and blood lipids in mice.

[0108] Fresh blood was collected from mice and centrifuged at 3000×g for 15 min. The colorless serum supernatant was collected, and the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), total cholesterol (TC), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) in the mouse serum were measured using a fully automated biochemical analyzer. The results are shown in Table 3.

[0109] Table 3. Effects of each drug group on liver function and four blood lipid indicators in mice.

[0110]

[0111] Note: # indicates comparison with the blank control group, #p<0.05, ##p<0.01; * indicates comparison with the model group, *p<0.05, **p<0.01.

[0112] The experimental results in Table 3 show that, compared with the blank control group, the serum levels of ALT, AST, TG, TC, and LDL-C in the model group mice were significantly increased (p<0.01). Compared with the model group, the serum levels of ALT, AST, TG, TC, and LDL-C in experimental groups 1-9 (compositions 1-9) were significantly decreased (p<0.01 and p<0.05), and the serum levels of ALT, AST, TG, TC, and LDL-C in experimental groups 10-15 (single herbal combination) were also decreased, but the differences were not statistically significant. The decrease was most significant in composition 1, suggesting that the traditional Chinese medicine in composition 1 has a synergistic effect and the optimal compatibility ratio. Compared with the control group, the improvement effect of compositions 1-3 was significantly better than that of the traditional Chinese medicine compositions in the control group.

[0113] 4. Determination of triglycerides and total cholesterol in mouse liver

[0114] Sample pretreatment (triglycerides): Weigh 100 mg of liver tissue and homogenize with 1 mL of aqueous solution containing 5% NP-40. Slowly heat the sample in a water bath to 80℃~100℃ for 2 min~5 min, then cool to room temperature. Repeat the heating and cooling process once to dissolve all triglycerides. Centrifuge at high speed for 2 min to remove insoluble matter. Dilute the sample 10-fold with dH2O before measurement, and then measure according to the instructions of the triglyceride quantitative analysis kit.

[0115] Sample pretreatment (total cholesterol): Weigh 10 mg of liver tissue and homogenize with 200 μL of a mixture (chloroform:isopropanol:NP-40 = 7:11:0.1). Centrifuge at 15000 ×g for 5-10 min. Transfer all organic phase to a new EP tube and air dry at 50°C to remove chloroform. Vacuum the sample for 30 min to remove trace amounts of organic solvent. Dissolve lipids in 200 μL of Cholesterol Assay Buffer from the kit, sonicate or vortex to homogenize, and then measure according to the instructions of the total cholesterol quantification kit.

[0116] Table 4. Effects of each drug group on liver triglycerides and total cholesterol in mice.

[0117]

[0118] The experimental results in Table 4 show that, compared with the blank control group, the liver TG and TC levels of mice in the model group were significantly increased (p<0.01). Compared with the model group, the liver TG and TC levels of mice in experimental groups 1-9 (compositions 1-9) were significantly decreased (p<0.01 and p<0.05), with the most significant decrease in experimental group 1 (composition 1), suggesting that the various Chinese herbal medicines in composition 1 have a synergistic effect and the optimal compatibility ratio, resulting in a significantly better effect than the individual application of any component in the formula. Compared with the control group Chinese herbal medicine compositions that replaced one Chinese herbal medicine and had different ratios, compositions 1-3 showed a significantly improved effect in reducing TG and TC.

[0119] (vi) Mouse liver fat expression detection experiment:

[0120] After the indicator tests were completed in mice, liver tissue was collected, fixed in 4% paraformaldehyde, embedded in paraffin, and cut into 4μm thick sections. Hematoxylin-eosin (HE) staining was performed, and the pathological changes in the liver tissue of each group of mice were observed under a light microscope. The results are shown in [Table missing]. Figure 1 In addition, fresh liver tissue was rapidly frozen in liquid nitrogen to prepare frozen sections, routinely stained with Oil Red O, and then immediately observed under an optical microscope. The results are shown in [Figure number missing]. Figure 2 .

[0121] according to Figure 1 and Figure 2 HE staining and Oil Red O staining results showed that in the normal group, intact hepatocytes and nuclei were visible, with the hepatocytes arranged tightly and orderly, and the structure clear and intact. In the model group, the hepatocytes of mice exhibited abnormal morphology, loose arrangement, and numerous fat vacuoles; Oil Red O staining revealed a large number of fat droplets, indicating the successful establishment of the non-alcoholic fatty liver disease mouse model. Compared with the model group, the degree of hepatocyte steatosis and the number of fat droplets were significantly reduced in the composition 1 group. This indicates that composition 1 can improve the liver pathological state in mice with non-alcoholic fatty liver disease.

[0122] Example 2: Safety Evaluation

[0123] 1. Acute toxicity test

[0124] Forty healthy mice, half male and half female, weighing 22±2g, were selected. After fasting for 12 hours, they were administered the traditional Chinese medicine composition 1 prepared in Example 1 at a dose of 21.36g / kg by gavage. After administration, they were fasted for another 4 hours before being fed. The mice were observed for 14 consecutive days, and their physical signs, weight, food intake, poisoning and mortality were recorded. The results are shown in Tables 5 and 6.

[0125] 2. Subchronic toxicity test

[0126] Forty healthy mice, half male and half female, weighing 22±2g, were selected. After fasting for 12 hours, they were administered the traditional Chinese medicine composition 1 prepared in Example 1 at a dose of 21.36g / kg by gavage. After administration, they were fasted for another 4 hours before being fed. The administration was repeated for 35 days. The mice's physical signs, weight, food intake, poisoning and mortality were recorded for 35 days. The results are shown in Table 7.

[0127] Table 5. Results of mouse weight measurement in acute toxicity experiment

[0128]

[0129] Table 6 Results of Acute Toxicity Test: Food Intake Measurement

[0130]

[0131] Table 7 Results of 35-day subchronic toxicity test

[0132]

[0133] The experimental results in Table 5-7 show that during the experimental observation period, the animals in each group had normal eating and activity, good growth, and no signs of poisoning or death were observed, indicating that the traditional Chinese medicine composition provided by this invention has good safety.

[0134] The above description of the technical solution of this invention further illustrates the beneficial effects of this invention. The formula of this invention deeply embodies the concept of "medicine and food sharing the same origin," meaning that it can be used as both medicine and food. The medicinal and food-based traditional Chinese medicine formula of this invention can further reduce safety risks associated with long-term medication and improve patient compliance. Medicinal and food-based traditional Chinese medicines are "components" for building a healthy China and developing the big health industry, and are the material basis for the human health community to face chronic diseases, possessing significant potential value in the treatment of chronic diseases.

[0135] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details of the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solutions of the present invention. These simple modifications all fall within the protection scope of the present invention. It should also be noted that the various specific technical features described in the above embodiments can be combined in any way without contradiction. To avoid unnecessary repetition, the present invention will not describe the various possible combinations separately. Furthermore, various different embodiments of the present invention can also be arbitrarily combined, as long as they do not violate the spirit of the present invention, they should also be considered as the content disclosed by the present invention.

Claims

1. A traditional Chinese medicine composition, characterized in that, The raw materials include the following parts by weight: 15-25 parts raw hawthorn, 10-20 parts dried tangerine peel, 10-15 parts cassia seed, 8-12 parts salvia miltiorrhiza, 10-20 parts jujube seed, and 10-20 parts raw licorice.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, It is prepared from the following parts by weight of raw materials: 18 parts raw hawthorn, 15 parts dried tangerine peel, 13 parts cassia seed, 10 parts salvia miltiorrhiza, 12 parts jujube seed, and 15 parts raw licorice.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, It is prepared from the following raw materials in parts by weight: 18 parts raw hawthorn, 12 parts dried tangerine peel, 13 parts cassia seed, 10 parts salvia miltiorrhiza, 10 parts jujube seed, and 10 parts raw licorice.

4. The traditional Chinese medicine composition according to claim 1, characterized in that, It is prepared from the following raw materials in parts by weight: 18 parts raw hawthorn, 18 parts dried tangerine peel, 13 parts cassia seed, 10 parts salvia miltiorrhiza, 20 parts jujube seed, and 20 parts raw licorice.

5. A method for preparing the traditional Chinese medicine composition according to any one of claims 1 to 4, characterized in that, Includes the following steps: Raw hawthorn, dried tangerine peel, cassia seed, salvia miltiorrhiza, jujube seed, and raw licorice were mixed, extracted with solvent, and the extract was collected and concentrated. The solvent is water or an aqueous solution of ethanol.

6. The preparation method according to claim 5, characterized in that, The volume fraction of the ethanol-water solution is 10%~95%; And / or, the extraction is a heating reflux extraction; And / or, the extraction is performed 2 to 3 times; And / or, for each extraction, the ratio of the solvent to the total weight of the raw material is (5~20):1, expressed in mL:g.

7. A traditional Chinese medicine extract, characterized in that, It is prepared by the preparation method according to any one of claims 5 or 6, or by extraction from the traditional Chinese medicine composition according to any one of claims 1 to 4.

8. The use of the traditional Chinese medicine composition according to any one of claims 1 to 4 or the traditional Chinese medicine extract according to claim 7 in the preparation of a medicament for the prevention and treatment of non-alcoholic fatty liver disease.

9. The application according to claim 8, characterized in that, The prevention and treatment of non-alcoholic fatty liver disease includes: Reduce weight and / or liver weight; And / or, reduce the level of at least one of the following indicators in serum: alanine aminotransferase, aspartate aminotransferase, triglycerides, total cholesterol, and low-density lipoprotein cholesterol; And / or, reduce the levels of triglycerides and / or total cholesterol in liver tissue; And / or, improve the pathological state of hepatic steatosis.

10. A drug for the prevention and treatment of non-alcoholic fatty liver disease, characterized in that, It includes the traditional Chinese medicine composition according to any one of claims 1 to 4 or the traditional Chinese medicine extract according to claim 7, as well as pharmaceutically acceptable excipients.

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