Traditional Chinese medicine component compound preparation for treating metabolism-related fatty liver disease and application of traditional Chinese medicine component compound preparation

Abstract

The invention discloses a traditional Chinese medicine component compound preparation for treating metabolism-related fatty liver diseases and application of the traditional Chinese medicine component compound preparation. The effective components of the traditional Chinese medicine component compound preparation comprise alisol B and atractylenolide III. According to the invention, the synergistic interaction mechanism of alisol B and atractylenolide III is disclosed for the first time, and the limitation of multi-component mixing of a traditional compound is broken through. After the two components are combined according to a specific ratio, the effects of improving liver functions and lowering lipid are remarkably improved compared with those of an original formula, the scientific breakthrough of formula reduction and effect improvement is achieved, the problems that compound components are complex and quality standards are difficult to control are solved, and the traditional Chinese medicine composition has no common side effects of weight gain and other western medicines and has unique advantages in treatment of metabolism-related fatty liver diseases.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and specifically designs a compound preparation of traditional Chinese medicine components for treating metabolism-related fatty liver disease and its application. Background Art

[0002] A definitive diagnosis of metabolic associated fatty liver disease (MAFLD) still requires excluding other causes of chronic liver disease, such as excessive alcohol consumption. MAFLD, formerly known as nonalcoholic fatty liver disease (NAFLD), has a global prevalence of up to 25%, severely posing a health threat to humans. To date, no medications have been approved for its treatment in the United States or the European Union. Unhealthy lifestyles, such as prolonged sedentary behavior and inactivity, as well as dietary habits characterized by excessive calorie intake and an irrational diet, are closely associated with the increasing incidence of MAFLD. Modern medical treatments for MAFLD primarily utilize drugs that protect the liver, lower enzyme levels, lower lipids, and modulate the immune system. While these diverse approaches are associated with significant adverse reactions, Traditional Chinese Medicine (TCM) offers advantages in improving clinical symptoms and reducing adverse reactions, combining a holistic approach with syndrome differentiation and treatment. However, traditional Chinese medicine formulas, due to their complex and unclear composition, pose challenges in formulation control and stability, a challenge that urgently needs to be addressed in the modernization of TCM. Developing safe, effective, and materially definitive traditional Chinese medicine (TCM) active ingredient compounds for the prevention and treatment of metabolic-related fatty liver disease (MAFLD) is crucial for the development of effective TCM compound formulations that eliminate the dross and retain the essence. This application aims to provide a TCM compound formulation with defined active ingredients and minimal toxicity and side effects, effectively combating MAFLD, with the goal of achieving a scientific breakthrough in reducing prescriptions and increasing efficacy. Summary of the Invention

[0003] In response to the problems existing in the prior art, the present invention aims to provide a compound preparation of traditional Chinese medicine components for treating metabolic-related fatty liver disease, which is specifically achieved through the following technical solutions:

[0004] The first aspect of the present invention provides a compound preparation of traditional Chinese medicine components for treating metabolism-related fatty liver disease, wherein the effective components of the compound preparation of traditional Chinese medicine consist of alismatol B and atractylodes lactone III.

[0005] Furthermore, the mass ratio of alismatol B to atractylodes lactone III is 40:1-1:1.

[0006] Furthermore, Alisol B is derived from the traditional Chinese medicine Alisma orientalis and can be obtained through conventional extraction methods or commercial purchase.

[0007] Furthermore, Atractylenolide III is derived from the traditional Chinese medicine Atractylodes macrocephala, and can be obtained through conventional extraction methods or commercial purchase.

[0008] Furthermore, the compound preparation also contains other pharmaceutically acceptable excipients, and the compound preparation is in any clinically acceptable dosage form, such as tablets, granules, capsules and other oral solid preparations.

[0009] The second aspect of the present invention provides the use of a traditional Chinese medicine compound preparation in the preparation of a drug for treating fatty liver disease.

[0010] Furthermore, the fatty liver disease is metabolism-related fatty liver disease induced by a high-fat diet.

[0011] Furthermore, the application is shown to significantly reduce the serum AST level in high-fat mice.

[0012] This invention reveals for the first time the synergistic mechanism of alismatol B and atractylodes lactone III, overcoming the limitations of traditional compound formulas, which often involve multiple components. When combined in a specific ratio, the two significantly enhance liver function and lipid-lowering effects compared to the original formula, achieving a scientific breakthrough in "reducing the formula while increasing efficacy." This overcomes the complex composition and difficult quality control issues of compound formulas, and eliminates common side effects of Western medicine, such as weight gain, offering unique advantages in the treatment of metabolic-related fatty liver disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 This is the effect of Xiezhuo Tiaozhi prescription on improving MAFLD ( n=8);

[0014] Figure 2 The effect of Xiezhuo Tiaozhi prescription on lipid accumulation in the liver of MAFLD mice ( n=8). DETAILED DESCRIPTION

[0015] The present invention is further described below in conjunction with specific embodiments to facilitate a better understanding of the present technical solution.

[0016] Example 1: Screening the optimal component formula of the Xiezhuo Tiaozhi formula for improving hepatic lipid accumulation in a high-fat diet-induced metabolic-related fatty liver disease mouse model using a uniform design method

[0017] 1. Materials

[0018] Experimental animals: 80 SPF-grade male C57BL / 6 mice, weighing (25±2) g, were purchased from Shanghai Slake Co., Ltd. The experimental animals were housed at the Experimental Animal Center of Zhejiang Chinese Medical University.

[0019] Main drugs and reagents:

[0020] Alismatol B (Cat. No. 230609), Atractylodes lactone III (Cat. No. 230908), and Pachymic acid (Cat. No. 230523) were purchased from Shanghai Ronghe Biological Co., Ltd. The original Xiezhu Tiaozhi formula (Zhiqi, stir-fried Atractylodes macrocephala, Poria cocos, raw hawthorn, lotus leaf, and stir-fried Citrus aurantium) was purchased from Zhejiang Chinese Medical University Chinese Medicine Piece Co., Ltd. Based on previous studies, a dosing concentration of 4.16 g / kg / day was formulated. Polyene phosphatidylcholine (Cat. No. H2005901) was purchased from Beijing Sanofi Pharmaceutical Co., Ltd. High-fat feed (Cat. No. TP26300, composition: 10% lard + 2% cholesterol + 88% basal feed) was purchased from Nanjing Trophy Feed Technology Co., Ltd.

[0021] 2. Methods

[0022] 2.1 Establishment of a mouse model of metabolic-related fatty liver disease

[0023] According to the existing technology, a NAFLD mouse model was established by feeding mice with a high-fat diet (10% lard + 2% cholesterol + 88% basic diet) for 12 weeks.

[0024] 2.2 Uniform design chicken dosing regimen

[0025] The uniform design method was used, with the three effective components or ingredients in the prescription as the investigation factors, and X1, X2, and X3 respectively representing the three effective components of the traditional Chinese medicine in the Xiezhuo Tiaozhi prescription. According to the previous results and combined with the literature, the dosage of alisamol B was 15-100 mg / kg, the dosage of atractylodes lactone III was 0.6-60 mg / kg, and the dosage of pachymic acid was 1-50 mg / kg. The dosage of alisamol B (100 mg / kg) in the previous study was used as level 6, the dosage of atractylodes lactone III (40 mg / kg) was used as level 5, and the dosage of pachymic acid (30 mg / kg) was used as level 6. According to the U6*(64) table (Table 1) and the usage table (Table 2), the 1st, 2nd, and 3rd columns were selected to obtain the uniform design table of the three drugs.

[0026] The experimental scheme of the three-drug combination is shown in Table 3, and the daily dosage of the effective components of the three Chinese herbal medicines is shown in Table 4.

[0027] At the 8th week of high-fat diet feeding, mice were given different doses of drugs by gavage, once a day, for four consecutive weeks. The blank group and the model group were gavaged with equal volume of 0.5% CMC-Na. At the 12th week of continuous gavage, samples were collected after anesthesia and the corresponding indicators were detected.

[0028] Table 1U6*(64)

[0029]

[0030] Table 2U6*(64) table usage table

[0031]

[0032] Table 3 Trial plan for the combination of ABC drugs

[0033]

[0034] Table 4 Uniform design experimental plan (daily dosage (mg / kg body weight)

[0035]

[0036] 2.3 Blood biochemical test and liver TG and TC test

[0037] The levels of ALT, AST, TC, and TG in mouse serum were detected using a fully automatic blood biochemical analyzer, and the TG and TC indicators in mouse liver tissue were detected using Nanjing Jiancheng kits.

[0038] 2.4 Histopathological observation

[0039] H&E staining was used to observe the pathological changes of mouse liver tissue; Oil red O staining was used to observe the lipid accumulation level in mouse liver tissue.

[0040] 2.5 Statistical analysis

[0041] SPSS 21.0 software was used for statistical analysis. If the data met the criteria of normality and homogeneity of variance, one-way ANOVA analysis of variance was performed, and pairwise comparisons were performed using the LSD-t test, with data presented as ±s. If the data did not meet the criteria of homogeneity of variance and normality, a nonparametric test for multiple independent samples (Kruskal-Wallis H test) was used. SPSS performed multivariate stepwise regression analysis on the uniform design results.

[0042] 3. Results

[0043] 3.1 The uniform design method was used to investigate the effects of the compound formula of purging turbidity and regulating lipids on serum liver function and blood lipid levels in MAFLD mice

[0044] The results are as follows Figure 1As shown in the figure, (A) the effect of the Xiezhuo Tiaozhi formula components and compound on the serum ALT of mice; (B) the effect of the Xiezhuo Tiaozhi formula components and compound on the serum AST of mice; (C) the effect of the Xiezhuo Tiaozhi formula components and compound on the serum TG of mice; (D) the effect of the Xiezhuo Tiaozhi formula components and compound on the serum TC of mice; (E) the effect of the Xiezhuo Tiaozhi formula components and compound on the serum TG of mice; (F) the effect of the Xiezhuo Tiaozhi formula components and compound on the liver TG of mice; (G) H&E staining to observe the histopathology of mouse liver (200×); (H) Oil red O staining to observe the lipid accumulation level in mouse liver (200×). Note: ND: normal diet group; HFD: high-fat diet group; JY1-6: uniform design 1-6 groups; XZTZ: original formula of Xiezhuo Tiaozhi group; YSF: Yishanfu group, i.e., polyene phosphatidylcholine group. Compared with the ND group, **P<0.01; compared with the HFD group, # P<0.05, ## P<0.01.

[0045] Depend on Figure 1 All components of the Xiezhuo Tiaozhi formula significantly reduced serum ALT and AST levels in mice induced by high fat diet, with JY1 showing significantly better efficacy than the original formula. Furthermore, JY1, JY2, JY5, and JY6, components of the Xiezhuo Tiaozhi formula, significantly reduced serum triglyceride (TG) levels in mice induced by high fat diet. JY1-5, components of the Xiezhuo Tiaozhi formula, significantly reduced serum TG levels. The components of the Xiezhuo Tiaozhi formula, JY1, JY5, and JY6, significantly reduced liver TG levels in mice, with JY1 showing significantly better efficacy than the original formula. All components of the Xiezhuo Tiaozhi formula, JY1-JY6, significantly reduced liver TG levels in mice, with JY6 showing significantly better efficacy than the original formula. H&E staining revealed varying degrees of steatosis and vacuolation in the liver tissue of mice in the high fat diet group. Degenerated hepatocytes increased in size, with numerous fat vacuoles appearing in the cytoplasm. Nuclei in some cells were seen extruding from the cell membrane, and a small number of inflammatory cells were observed in the necrotic areas. The uniform design of the Xiezhuo Tiaozhi formula group, the Xiezhuo Tiaozhi formula group, and the positive drug group (polyene phosphatidylcholine) significantly improved liver steatosis in mice. Oil Red O staining revealed that the model group mice had abundant lipid droplet deposition in their liver tissues. All Xiezhuo Tiaozhi formula groups effectively improved lipid accumulation in the liver tissues of mice. Using liver TG as the key indicator, a regression equation was calculated, yielding Y = 0.15-2.990×10-7X12 + 1.679×10-5X2. The regression equation suggests that the optimal theoretical compatibility ratio of the Xiezhuo Tiaozhi formula components and compound is achieved when the dose of Alisma alcohol B is maximized and the dose of Atractylodes lactone III is minimized. Further pharmacodynamic experiments are needed to verify the theoretical optimal compatibility of the Xiezhuo Tiaozhi formula components and compound.

[0046] Example 2: Verification of the effect of the compound formula for purging turbidity and regulating lipids on lipid accumulation in the liver of MAFLD mice

[0047] The results are as follows Figure 2 As shown in the figure, (A) Effects of the Xiezhuo Tiaozhi Formula components and compound on serum ALT in mice; (B) Effects of the Xiezhuo Tiaozhi Formula components and compound on serum AST in mice; (C) Effects of the Xiezhuo Tiaozhi Formula components and compound on serum TG in mice; (D) Effects of the Xiezhuo Tiaozhi Formula components and compound on serum TC in mice; (E) Effects of the Xiezhuo Tiaozhi Formula components and compound on serum TG in mice; (F) Effects of the Xiezhuo Tiaozhi Formula components and compound on liver TG in mice; (G) H&E staining of mouse liver histopathology (200×); (H) Oil Red O staining of mouse liver lipid accumulation (200×). Note: ND: normal diet group; HFD: high-fat diet group; JY: uniform design group; HG: regression equation group; XZTZ: Xiezhuo Tiaozhi group; YSF: Yishanfu group. Compared with the ND group, **P < 0.01; compared with the HFD group, #P < 0.05, ##P < 0.01.

[0048] Depend on Figure 2 The JY and HG groups significantly improved serum ALT levels in mice induced by a high-fat diet. The HG group significantly reduced serum AST levels in mice induced by a high-fat diet, with efficacy comparable to that of the Xiezhuo Tiaozhi formula group. The HG group significantly improved serum TG levels in mice, with the best efficacy. Furthermore, the JY group improved serum TC in mice induced by a high-fat diet, but no significant difference was found between the HG group and the model group. Both the JY and HG groups effectively improved liver TC and TG levels in NAFLD mice, with efficacy comparable to that of the Xiezhuo Tiaozhi formula and Yishanfu groups. H&E staining revealed that liver tissues in the high-fat diet model group displayed varying degrees of steatosis and vacuolation, with enlarged degenerated hepatocytes and numerous fat vacuoles within the cytoplasm. The JY and HG groups significantly improved steatosis and vacuolation in liver tissues, with superior efficacy to the Xiezhuo Tiaozhi formula and Yishanfu groups. Oil Red O staining revealed significant lipid droplet accumulation in the liver tissues of mice in the high-fat diet group. The JY and HG groups effectively reduced lipid accumulation induced by high-fat diets, with superior efficacy to the original formula group. In summary, the HG group significantly improved hepatic lipid accumulation in MAFLD mice, with a superior effect compared to the original formula. This result demonstrates the reliability of the Xiezhuo Tiaozhi formula component ratios derived from the regression equation. Therefore, the Xiezhuo Tiaozhi formula component compound derived from the regression equation was named "AA," and its optimal dosage was 100 mg / kg of alismatol B and 2.5 mg / kg of atractylodes lactone III.

[0049] The compound preparation of the present invention can significantly reduce liver damage, liver tissue triglyceride levels, and alleviate liver tissue pathological damage and the degree of hepatic steatosis in mice with metabolic-related fatty liver disease models, with effects superior to those of either alone or the original compound. This compound preparation can effectively improve metabolic-related fatty liver disease and prevent the progression of the disease, and can be used to treat and prevent metabolic-related fatty liver disease, chronic liver damage, and other conditions. Compared to traditional Chinese medicine compounds, this compound preparation has advantages such as clear ingredients, controllable quality, high stability, and clear efficacy, and explains, from a certain perspective, the effective substance components of Chinese medicine compounds for treating metabolic-related fatty liver disease.

Claims

1. A compound preparation of traditional Chinese medicine components for treating metabolic-related fatty liver disease, characterized in that: The effective components of the traditional Chinese medicine compound preparation are composed of alismatol B and atractylodes lactone III.

2. The compound preparation of traditional Chinese medicine components for treating metabolic-related fatty liver disease according to claim 1, characterized in that: The mass ratio of the alismatol B to the atractylodes lactone III is 40:1-1:

1.

3. The compound preparation of traditional Chinese medicine components for treating metabolic-related fatty liver disease according to claim 1, characterized in that: The compound preparation also contains other pharmaceutically acceptable excipients, and the compound preparation is in any clinically acceptable dosage form.

4. Use of the Chinese herbal compound preparation according to any one of claims 1 to 3 in the preparation of a medicament for treating fatty liver disease.

5. The use according to claim 4, characterized in that The fatty liver disease is a metabolic-related fatty liver disease induced by a high-fat diet.

6. The use according to claim 5, characterized in that The application is shown to significantly reduce the serum AST level in high-fat mice.

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