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

Abstract

The present application belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for preventing or treating fatty liver disease, and a preparation method and application thereof. The traditional Chinese medicine composition is made of the following raw materials in parts by weight: 5-30 parts of atractylodes, 5-30 parts of prepared cyperus, 5-30 parts of dodder, 5-30 parts of raspberry, 5-30 parts of medlar, 5-30 parts of mistletoe, 5-30 parts of fried continuation, 5-30 parts of salt eucommia, 5-30 parts of salvia miltiorrhiza, and 5-30 parts of chuanxiong. The present application adopts the method of syndrome differentiation and treatment, and the best formula is screened through a large number of experiments. The treatment is based on the spleen, and the principle is to soothe the liver and gallbladder, remove phlegm and blood stasis, and invigorate the spleen and kidney, so as to play the role of preventing and treating fatty liver disease. The animal experiment results show that the traditional Chinese medicine composition has a significant therapeutic effect on fatty liver disease.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine composition for the prevention or treatment of fatty liver disease, its preparation method, and its uses. Background Technology

[0002] Non-alcoholic fatty liver disease (NAFLD) is a clinicopathological syndrome characterized primarily by hepatocyte steatosis. Its scope includes simple fatty liver, non-alcoholic steatohepatitis (NASH), which can further progress to liver fibrosis and cirrhosis, and even lead to hepatocellular carcinoma. Epidemiological studies show that NAFLD is one of the most prevalent liver diseases globally, with a global prevalence of 25.24%, and the prevalence is increasing annually. NASH has a high detection rate in NAFLD patients with concurrent metabolic syndrome and type 2 diabetes. NASH is one of the leading causes of cirrhosis; the incidence of cirrhosis in NASH patients is as high as 15%-25% within 10-15 years. NASH represents a crucial stage in the progression from simple fatty liver to liver fibrosis and cirrhosis, and is also a significant breakthrough in the study of the pathogenesis of chronic liver diseases. The goal of NASH treatment is to slow, stop, and reverse the progression of NASH, improve clinical outcomes, including reducing the incidence of cirrhosis and its complications, reducing the need for liver transplantation, improving survival rates, and improving quality of life.

[0003] The pathogenesis of non-alcoholic fatty liver disease (NAFLD) is complex, and it is closely related to obesity, type 2 diabetes, hyperlipidemia, and metabolic syndrome. Currently, treatment primarily involves diet and exercise control, with commonly used medications including silymarin, ursodeoxycholic acid, polyene phosphatidylcholine, reduced glutathione, bicyclol, and biphenyl diester. While these have some effect on NAFLD treatment, the use of single drugs has limitations and the benefits are limited. In Traditional Chinese Medicine (TCM), NAFLD is classified as "liver obstruction," "accumulation," and "hypochondriac pain." The disease primarily affects the liver and spleen, often due to improper diet, overwork, emotional disturbances, leading to spleen dysfunction, liver qi stagnation, or liver-spleen imbalance. Prolonged illness can affect the kidneys, causing kidney essence deficiency and impaired kidney function in water metabolism. The pathological products of NAFLD are mostly phlegm, dampness, blood stasis, and heat, making it a complex disease involving both deficiency and excess.

[0004] Chinese patent CN115869358B discloses a traditional Chinese medicine composition for treating non-alcoholic fatty liver disease, which is made from the following raw materials in parts by weight: 5-25 parts of raw Astragalus membranaceus, 3-15 parts of Atractylodes lancea, 3-15 parts of Polygonum cuspidatum, 3-15 parts of Polygonum hydropiper, 3-15 parts of Lycopus lucidus, and 2-10 parts of Citrus reticulata peel. In this formula, raw Astragalus membranaceus invigorates Qi and strengthens the spleen, promoting the elimination of dampness and phlegm, serving as the principal herb. Atractylodes lancea dries dampness and strengthens the spleen, while Polygonum cuspidatum promotes diuresis and eliminates phlegm, enhancing the spleen-strengthening and phlegm-eliminating effects. Polygonum hydropiper clears heat and promotes diuresis, while Lycopus lucidus invigorates blood, removes blood stasis, and reduces swelling, further enhancing the heat-clearing and dampness-eliminating effects. Citrus reticulata peel strengthens the spleen, dries dampness, and also regulates Qi and resolves phlegm, serving as the guiding herb. The combined effects of these herbs invigorate the spleen, eliminate phlegm, clear heat, and promote diuresis, thus strengthening the spleen, clearing damp-heat, reducing phlegm and dampness, and ultimately leading to recovery.

[0005] Chinese patent CN114558074B discloses a traditional Chinese medicine composition for treating metabolic-related fatty liver disease (MAFLD). The composition includes: 10-20g of stir-fried Citrus aurantium (braised), 10-20g of Scutellaria baicalensis (processed with wine), 30-60g of Sedum sarmentosum, 10-20g of Codonopsis pilosula, 15-30g of Astragalus membranaceus, 10-20g of stir-fried Atractylodes macrocephala (braised), 10-20g of Poria cocos, 6-12g of Bupleurum chinense (processed with vinegar), 10-20g of stir-fried Paeonia lactiflora (stir-fried), 10-20g of Cyperus rotundus (processed with vinegar), 10-20g of Citrus reticulata peel, 10-20g of Citrus reticulata peel (processed with vinegar), 10-20g of Pinellia ternata (processed), 10-20g of Nelumbo nucifera leaf, 10-20g of Alisma plantago-aquatica, 10-20g of Crataegus pinnatifida (processed with charred), 10-20g of Liquidambar formosana (processed), and 3-6g of Glycyrrhiza uralensis (processed with roasted). This formula treats the spleen, focusing on resolving phlegm and dampness to promote weight loss and alleviate MAFLD.

[0006] There are many traditional Chinese medicine compositions for treating fatty liver disease, but they all have some drawbacks, such as too many ingredients, difficulty in sourcing raw materials, high prices, and unclear efficacy and poor treatment results. Therefore, there is an urgent need for a drug that is effective in treating fatty liver disease, has fewer ingredients, has widely available raw materials, and is easy to prepare. Summary of the Invention

[0007] The purpose of this invention is to overcome the shortcomings of the prior art by screening through a large number of experiments and combining clinical experience, and using the theory of syndrome differentiation and treatment to screen out traditional Chinese medicine compositions that can prevent or treat fatty liver disease, as well as their preparation methods and uses.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] In a first aspect, the present invention provides a traditional Chinese medicine composition for the prevention or treatment of fatty liver disease, wherein the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 5-30 parts of Atractylodes lancea, 5-30 parts of processed Cyperus rotundus, 5-30 parts of Cuscuta chinensis, 5-30 parts of Rubus idaeus, 5-30 parts of Lycium barbarum, 5-30 parts of Viscum album, 5-30 parts of stir-fried Dipsacus asper, 5-30 parts of salt-processed Eucommia ulmoides, 5-30 parts of Salvia miltiorrhiza, and 5-30 parts of Ligusticum chuanxiong.

[0010] Alternatively, in the above-mentioned traditional Chinese medicine composition, by weight, the traditional Chinese medicine composition is made from the following raw materials: 5-15 parts of Atractylodes lancea, 5-15 parts of processed Cyperus rotundus, 10-20 parts of Cuscuta chinensis, 10-20 parts of Rubus idaeus, 5-15 parts of Lycium barbarum, 10-20 parts of Viscum album, 10-20 parts of stir-fried Dipsacus asper, 5-15 parts of salt-processed Eucommia ulmoides, 10-20 parts of Salvia miltiorrhiza, and 5-15 parts of Ligusticum chuanxiong.

[0011] As an optional method, in the above-mentioned traditional Chinese medicine composition, by weight, the traditional Chinese medicine composition is made from the following raw materials: 12 parts of Atractylodes lancea, 10 parts of processed Cyperus rotundus, 15 parts of Cuscuta chinensis, 15 parts of Rubus idaeus, 10 parts of Lycium barbarum, 15 parts of Viscum album, 15 parts of stir-fried Dipsacus asper, 10 parts of salt-processed Eucommia ulmoides, 15 parts of Salvia miltiorrhiza, and 10 parts of Ligusticum chuanxiong.

[0012] In a second aspect, the present invention provides a method for preparing the traditional Chinese medicine composition described in the first aspect above. The preparation method includes the following steps: pulverizing Salvia miltiorrhiza, Cuscuta chinensis, Rubus idaeus, Loranthus parasiticus, stir-fried Dipsacus asper, Atractylodes lancea, processed Cyperus rotundus, Lycium barbarum, salt-processed Eucommia ulmoides, and Ligusticum chuanxiong, adding water for decoction, mixing the filtrate, and concentrating to obtain a concentrated solution.

[0013] As an optional method, in the above preparation method, the decoction is performed twice. After the first decoction is performed for 1-3 hours, the liquid is filtered to obtain a first filtrate. The residue is then added to water and decocted for another 1-3 hours, and filtered to obtain a second filtrate. The first and second filtrates are then combined.

[0014] Alternatively, in the above preparation method, the ratio of material to liquid in the first decoction and the second decoction is 1:4, the decoction time for the first decoction and the second decoction is 1.5 hours, and the water used in the first decoction and the second decoction is distilled water; preferably, the concentration is vacuum concentration.

[0015] In a third aspect, the present invention provides the use of the traditional Chinese medicine composition described in the first aspect above, or the traditional Chinese medicine composition prepared by the preparation method described in the second aspect above, in the preparation of a medicament for the prevention or treatment of fatty liver disease.

[0016] Alternatively, in the above-described uses, the fatty liver disease is non-alcoholic fatty liver disease.

[0017] Alternatively, in the above-described uses, the non-alcoholic fatty liver disease is selected from one or more of the following: simple fatty liver, polycystic ovary syndrome with fatty liver, non-alcoholic steatohepatitis (NASH), liver fibrosis, cirrhosis, or hepatocellular carcinoma.

[0018] Preferably, the non-alcoholic fatty liver disease is non-alcoholic steatohepatitis.

[0019] Alternatively, in the above-mentioned uses, the drug has the effects of soothing the liver and gallbladder, resolving phlegm and removing blood stasis, and strengthening the spleen and benefiting the kidneys.

[0020] Compared with the prior art, the present invention has the following advantages:

[0021] This invention employs a syndrome differentiation and treatment approach, selecting the optimal formula through extensive experimentation. It focuses on treating the spleen, employing the principles of soothing the liver and gallbladder, resolving phlegm and removing blood stasis, and strengthening the spleen and kidneys to prevent and treat fatty liver disease. Animal experiments demonstrate that the herbal composition of this invention has a significant therapeutic effect on fatty liver disease. Attached Figure Description

[0022] Figure 1 Effects of the lipid-regulating and liver-benefiting decoction on the body weight of mice with fatty liver. Compared with the control group, *** P < 0.001; compared with the model group, # P<0.05, ▲ P<0.05, ◇◇ P<0.05.

[0023] Figure 2 The effect of a lipid-regulating and liver-benefiting decoction on serum alanine aminotransferase (ALT) levels in fatty liver mice. Compared with the control group, ** P<0.01; compared with the model group, # P<0.05.

[0024] Figure 3 The effect of a lipid-regulating and liver-benefiting decoction on serum total cholesterol levels in mice with fatty liver. Compared with the control group, ** P<0.01; compared with the model group, # P<0.05.

[0025] Figure 4 The effect of a lipid-regulating and liver-benefiting decoction on triglyceride content in the liver tissue of mice with fatty liver. Compared with the control group, * P<0.05; compared with the model group, # P<0.05, △Δ△ P<0.001.

[0026] Figure 5 The effect of a lipid-regulating and liver-benefiting decoction on the total cholesterol content in the liver tissue of mice with fatty liver. Compared with the control group, **** P < 0.0001; compared with the model group, ### P<0.001.

[0027] Figure 6 Effects of Tiaozhi Yigan Decoction on pathological changes in liver tissue of fatty liver mice (×100). A: Control group; B: Model group; C: Tiaozhi Yigan Decoction I group; D: Tiaozhi Yigan Decoction II group; E: Wuzi Cangfu Guixiong Er Chen Decoction group.

[0028] Figure 7 Effects of Tiaozhi Yigan Decoction on pathological changes in liver tissue of fatty liver mice (×400). A: Control group; B: Model group; C: Tiaozhi Yigan Decoction Group I; D: Tiaozhi Yigan Decoction Group II; E: Wuzi Cangfu Guixiong Er Chen Decoction Group.

[0029] Figure 8 Effects of Tiaozhi Yigan Decoction on Liver Changes in Fatty Liver Mice. A: Control Group; B: Model Group; C: Tiaozhi Yigan Decoction Group I; D: Tiaozhi Yigan Decoction Group II; E: Wuzi Cangfu Guixiong Er Chen Decoction Group.

[0030] Figure 9 The effect of a lipid-regulating and liver-benefiting decoction on triglyceride content in liver tissue of mice with fatty liver due to polycystic ovary syndrome. Compared with the control group, * P<0.05; compared with the model group, # P<0.05.

[0031] Figure 10 Effects of Tiaozhi Yigan Decoction on liver histopathological changes in fatty liver mice with polycystic ovary syndrome (×100). A: Normal group; B: Model group; C: Tiaozhi Yigan Decoction group; D: Positive drug group.

[0032] Figure 11 Effects of Tiaozhi Yigan Decoction on liver histopathological changes in fatty liver mice with polycystic ovary syndrome (×400). A: Normal group; B: Model group; C: Tiaozhi Yigan Decoction group; D: Positive drug group.

[0033] Figure 12 Effects of Tiaozhi Yigan Decoction on ovarian histopathological changes in fatty liver mice with polycystic ovary syndrome (×40). A: Normal group; B: Model group; C: Tiaozhi Yigan Decoction group; D: Positive drug group.

[0034] Figure 13 Effects of a lipid-regulating and liver-benefiting decoction on MDA content in liver tissue of mice with fatty liver due to polycystic ovary syndrome. Compared with the control group, ** P<0.01; compared with the model group, # P<0.05, △ P<0.05. Detailed Implementation

[0035] The formula analysis and treatment principles of the traditional Chinese medicine composition described in this invention are as follows:

[0036] Fatty liver falls under the category of liver disorders in Traditional Chinese Medicine. The disease is located in the liver and is closely related to the spleen, stomach, and kidneys. The liver is yin in nature but yang in function. If the liver fails to regulate the flow of qi, the liver qi will overpower the spleen, leading to impaired spleen function and the production of pathological products such as phlegm, dampness, and blood stasis. The spleen and kidneys are the foundation of both innate and acquired health, and over time, both the spleen and kidneys will become deficient.

[0037] Pathogenesis: Liver qi stagnation, phlegm and blood stasis, and spleen and kidney deficiency.

[0038] Treatment principles: Soothe the liver and gallbladder, resolve phlegm and remove blood stasis, strengthen the spleen and benefit the kidneys.

[0039] Prepared Cyperus rotundus has a pungent, slightly bitter, slightly sweet, and neutral flavor. It enters the liver, spleen, and triple burner meridians and has the effects of soothing the liver and relieving depression, regulating qi and relieving chest tightness, and regulating menstruation and relieving pain. It is an excellent product for soothing the liver and regulating qi.

[0040] Salvia miltiorrhiza is bitter and slightly cold in nature, and enters the liver, heart and pericardium meridians. It has the effects of promoting blood circulation and relieving pain, removing blood stasis and cooling blood, nourishing blood and calming the mind, and can break up masses and remove lumps.

[0041] Chuanxiong (Ligusticum striatum) is pungent and warm in nature, and enters the liver, gallbladder, and pericardium meridians. It has the effects of promoting blood circulation, dispelling wind and relieving pain, and dispersing stagnant qi and breaking up blood stasis.

[0042] Goji berries are sweet and neutral in nature, and enter the liver, kidney, and lung meridians. They have the effects of nourishing the liver and kidneys, replenishing essence and moistening the lungs, and expelling pathogenic factors from the five internal organs.

[0043] Atractylodes lancea is pungent, bitter, and warm in nature, and enters the spleen and stomach meridians. It has the effects of drying dampness and strengthening the spleen, dispelling wind and eliminating dampness. Long-term use can lighten the body and prolong life.

[0044] Salt-processed Eucommia ulmoides is sweet and warm in nature, and enters the liver and kidney meridians. It has the effects of tonifying the liver and kidneys, strengthening tendons and bones, and nourishing essence and liver blood.

[0045] Cuscuta seed is pungent, sweet, and neutral in nature, and enters the liver and kidney meridians. It has the effects of tonifying yang and nourishing yin, consolidating essence and improving eyesight, and enters the blood aspect of the three yin meridians of the foot.

[0046] Raspberries are sweet, sour, and slightly warm in nature, and enter the liver and kidney meridians. They have the effects of regulating and nourishing the liver and kidneys, replenishing deficiency and consolidating essence. Their sweet and warm nature soothes and harmonizes the five internal organs.

[0047] Mistletoe is neutral and bitter in nature, and enters the liver and kidney meridians. It has the effects of tonifying the liver and kidneys, strengthening tendons and bones, and dispelling wind and dampness. It also tonifies kidney qi and balances yin and yang.

[0048] Stir-fried Dipsacus root is slightly warm, bitter, and pungent in nature. It enters the liver and kidney meridians and has the effects of tonifying the liver and kidneys, strengthening tendons and bones, and promoting blood circulation. It also promotes blood circulation, breaks up blood stasis, astringes blood, and replenishes deficiencies.

[0049] The combined effects of these herbs can fully exert their therapeutic functions of soothing the liver and gallbladder, resolving phlegm and removing blood stasis, and strengthening the spleen and kidneys, thereby regulating qi and blood, eliminating pathogenic factors, and relieving various symptoms.

[0050] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the invention.

[0051] For those without specific technologies or conditions indicated in the examples, follow the technologies or conditions described in the literature in this field or the product instructions. For reagents or instruments without the manufacturer indicated, they are all conventional products that can be obtained through regular channels.

[0052] Unless otherwise specified, the experimental methods in the following examples are all conventional methods. Unless otherwise specified, the test materials used in the following examples are all commercially available products.

[0053] Example 1: Therapeutic effect of Tiaozhi Yigan Decoction on simple fatty liver

[0054] 1. Experimental materials

[0055] 1.1 Experimental animals

[0056] 50 male C57BL / 6N mice, weighing 22 - 24 g, of SPF grade, were purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd., and the animal license number is SYXK(Zhe)-2021-0007.

[0057] All animals were housed in a barrier - level animal room during the experiment, with constant temperature and humidity, 12 h of light and 12 h of dark each day, standard feed, and free access to water.

[0058] 1.2 Animal feed

[0059] The high - fat diet (D12108C) was purchased from Research Diets, Inc. (USA), which contains 40% fat and 1.25% cholesterol.

[0060] 1.3 Drug preparation

[0061] Tiaozhi Yigan Decoction I (10 herbs, the traditional Chinese medicine composition of the present invention): Atractylodes lancea 12 g, Cyperus rotundus 10 g, Cuscuta chinensis 15 g, Rubus idaeus 15 g, Lycium barbarum 10 g, Viscum coloratum 15 g, Dipsacus asperoides 15 g, Eucommia ulmoides 10 g, Salvia miltiorrhiza 15 g, Ligusticum chuanxiong 10 g.

[0062] Tiaozhi Yigan Decoction II (8 herbs, comparative traditional Chinese medicine composition 1): Atractylodes lancea 12 g, Cyperus rotundus 10 g, Rubus idaeus 15 g, Lycium barbarum 10 g, Viscum coloratum 15 g, Salvia miltiorrhiza 15 g, Ligusticum chuanxiong 10 g, Astragalus membranaceus 15 g.

[0063] Wuzi Cangfu Guixiong Erchen Decoction (comparative traditional Chinese medicine composition 2): Lycium barbarum 10 g, Cuscuta chinensis 15 g, Rubus idaeus 10 g, Atractylodes lancea 10 g, Cyperus rotundus 10 g, Angelica sinensis 10 g, Ligusticum chuanxiong 10 g, Poria cocos 12 g, Citrus reticulata 10 g, Pinellia ternata 10 g, Crataegus pinnatifida 15 g, Aurantii Fructus Immaturus 10 g.

[0064] 1.4 Decoction preparation plan

[0065] First, soak each of the above-mentioned prescriptions in 1000mL of water for 2 hours. Then, heat the induction cooker at 1300W and boil the liquid. Once the liquid begins to boil, reduce the power to 120W and continue heating for 1.5 hours. Filter the liquid to obtain liquid a. Then, add another 1000mL of water and boil the liquid for 1.5 hours using the same method. Filter the liquid to obtain liquid b. Mix liquid a and liquid b and concentrate the liquid to 100mL using a rotary evaporator. Finally, refrigerate the liquid for later use.

[0066] 1.5 Animal grouping and intervention

[0067] After one week of acclimatization feeding, SPF-grade C57BL / 6N mice were randomly divided into 5 groups (n=10 per group): normal control group, fatty liver model group, Tiaozhi Yigan Tang I group (50g / kg / d), Tiaozhi Yigan Tang II group (50g / kg / d), and Wuzi Cangfu Guixiong Erchen Tang group (50g / kg / d). The normal control group was fed a standard diet, while the other groups were fed a high-fat diet. After 12 weeks of intervention, the mice were fasted for 12 hours, then euthanized by decapitation. Blood was collected, centrifuged (3,500 rpm, 4℃, 10 min), and stored at -80℃. Liver tissue was then rapidly dissected from the mice. A small portion of the liver tissue sample was fixed in 4% paraformaldehyde solution and stored at 4℃ for morphological observation; the remaining liver tissue sample was flash-frozen in liquid nitrogen and stored at -80℃ for subsequent indicator determination.

[0068] 2. Results and Analysis

[0069] 2.1 Effects of the Lipid-Regulating and Liver-Benefiting Decoction on Body Weight in Mice with Fatty Liver

[0070] Weight monitoring results of mice after 12 weeks of feeding are as follows Figure 1 As shown, compared with the normal diet control group, mice treated with a high-fat diet (HFD) showed a significant increase in body weight, with a significant difference starting in the fifth week (p<0.01); while after treatment with Tiaozhi Yigan Decoction and Er Chen Decoction, the weight gain trend of mice in the Tiaozhi Yigan Decoction group and the Wuzi Cangfu Guixiong Er Chen Decoction group slowed down significantly, with a significant difference starting in the fourth week compared with the model group.

[0071] 2.2 Effects of the Lipid-Regulating and Liver-Benefiting Decoction on Serum Detection Indicators in Fatty Liver Mice

[0072] Compared with the normal control group, the alanine aminotransferase (ALT) and total cholesterol (T-CHO) levels in the model group were significantly increased (p<0.01). After treatment with Tiaozhi Yigan Decoction I, ALT and T-CHO levels significantly decreased (p<0.05). After treatment with Tiaozhi Yigan Decoction II, ALT levels decreased but the difference was not significant, while T-CHO levels increased. After treatment with Wuzi Cangfu Guixiong Erchen Decoction, ALT levels decreased but the difference was not significant, while T-CHO levels increased. Figures 2-3 The results showed that Tiaozhi Yigan Decoction I could effectively reduce ALT and T-CHO levels in fatty liver mice. Compared with Tiaozhi Yigan Decoction II and Wuzi Cangfu Guixiong Erchen Decoction, Tiaozhi Yigan Decoction I had a more significant therapeutic effect on fatty liver caused by a high-fat diet.

[0073] 2.3 Effects of the Lipid-Regulating and Liver-Benefiting Decoction on Liver Lipids in Fatty Liver Mice

[0074] Compared with the normal control group, the triglyceride (TG) level in the model group was significantly increased (p<0.05). After treatment with Tiaozhi Yigan Decoction, the TG level in the Tiaozhi Yigan Decoction group was significantly decreased (p<0.05). The therapeutic effect of Tiaozhi Yigan Decoction II was better than that of Tiaozhi Yigan Decoction I. After treatment with Wuzi Cangfu Guixiong Erchen Decoction, the TG level decreased, but the difference was not statistically significant. Figure 4 The T-CHO level in the model group was significantly higher than that in the normal control group (p<0.0001). After treatment with Tiaozhi Yigan Decoction I, the T-CHO level in the Tiaozhi Yigan Decoction group significantly decreased (p<0.001). However, after treatment with Tiaozhi Yigan Decoction II and Wuzi Cangfu Guixiong Er Chen Decoction, the T-CHO level in these groups actually increased compared to the model group. Figure 5 The treatment results of Tiaozhi Yigan Decoction and Wuzi Cangfu Guixiong Er Chen Decoction show that Tiaozhi Yigan Decoction I can effectively reduce TG and T-CHO levels in fatty liver mice, and Tiaozhi Yigan Decoction II can effectively reduce TG levels in fatty liver mice. Compared with Wuzi Cangfu Guixiong Er Chen Decoction, Tiaozhi Yigan Decoction has a more significant therapeutic effect on fatty liver caused by a high-fat diet.

[0075] 2.4 Effects of the Lipid-Regulating and Liver-Benefiting Decoction on Pathological Changes in Liver Tissue of Mice with Fatty Liver

[0076] After HE staining of liver tissue from mice in each group, the control group showed intact and orderly hepatocyte morphology with no obvious fat vacuoles. The model group mice exhibited fatty degeneration and ballooning fatty degeneration, hepatocyte damage and necrosis, inflammatory cell infiltration, and numerous fat vacuoles. The Tiaozhi Yigan Decoction group significantly altered this lesion, with a marked reduction in fat vacuoles, demonstrating a significant therapeutic effect. Compared to the model group, the Wuzi Cangfu Guixiong Erchen Decoction group showed a reduction in fat vacuoles and a certain degree of therapeutic effect, but compared to the Tiaozhi Yigan Decoction group, the Tiaozhi Yigan Decoction showed better therapeutic effect, with Tiaozhi Yigan Decoction I showing better therapeutic effect than Tiaozhi Yigan Decoction II. The experimental results are as follows: Figure 6 and Figure 7 As shown.

[0077] 2.5 Effects of the Lipid-Regulating and Liver-Benefiting Decoction on Liver Changes in Fatty Liver Mice

[0078] To observe the overall size and color of the livers of C57BL / 6N mice, the modeling effect of a high-fat diet, and the therapeutic effects of Tiaozhi Yigan Decoction I, Tiaozhi Yigan Decoction II, and Wuzhi Cangfu Guixiong Erchen Decoction groups, the livers of mice in each group were immediately photographed and saved when taken out, as Figure 8 shown. The livers of mice in the normal control group were normal in size, bright red in color, and soft in texture; compared with the normal control group, the livers of the model group were abnormally enlarged, lighter yellow in color, and brittle in texture, with an oily luster and granular feeling visible on the liver surface to the naked eye; compared with the model group, the livers of mice in the Tiaozhi Yigan Decoction I group were close to those of the normal control group in color, the liver volume decreased, and the fatty degeneration improved; the livers of mice in the Tiaozhi Yigan Decoction II group also decreased in size to some extent, but were yellow in color, with an oily luster on the surface, and had a granular texture; the Wuzhi Cangfu Guixiong Erchen Decoction group was bright red in color, the liver volume was smaller than that of the model group, and slightly larger than that of the Tiaozhi Yigan Decoction I group. This shows that both Tiaozhi Yigan Decoction I and Wuzhi Cangfu Guixiong Erchen Decoction have therapeutic effects, and the therapeutic effect of Tiaozhi Yigan Decoction I is better.

[0079] Example 2: Therapeutic effect of Tiaozhi Yigan Decoction on polycystic ovary syndrome complicated with fatty liver 1. Experimental materials and methods

[0080] 1.1 Experimental animals

[0081] Forty C57BL / 6N mice, female, weighing 18 - 20 g, SPF grade, were purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd., with the animal license number SYXK(Zhe)-2021-0007.

[0082] All animals were housed in a barrier-level animal room during the experiment, with constant temperature and humidity, 12 hours of light and dark each day, standard feed, and free access to water.

[0083] 1.2 Animal feed

[0084] The high-fat feed (D12108C) was purchased from Research Diets, Inc., USA, which contained 40% fat and 1.25% cholesterol.

[0085] 1.3 Drug preparation

[0086] Tiaozhi Yigan Decoction I (10 herbs, the traditional Chinese medicine composition of the present invention): Atractylodes lancea 12 g, Cyperus rotundus 10 g, Cuscuta chinensis 15 g, Rubus idaeus 15 g, Lycium barbarum 10 g, Viscum coloratum 15 g, Dipsacus asperoides 15 g, Eucommia ulmoides 10 g, Salvia miltiorrhiza 15 g, Ligusticum chuanxiong 10 g.

[0087] Metformin (200 mg / kg): One tablet of metformin was taken and ground and mixed evenly with 20 mL of sodium carboxymethylcellulose.

[0088] Ethinylestradiol cyproterone acetate tablets (Diane, 0.2 mg / kg): Dissolve one Diane tablet in 100 mL of water and refrigerate for later use. 0.009 mL / g.

[0089] Androgen DHT (50 μg / μL): 2 mg DHT was diluted with 40 μL DMSO to 50 μg / μL, and then prepared with 1.96 mL corn oil to 1 μg / μL.

[0090] 1.4 Decoction Method

[0091] First, soak each of the above-mentioned prescriptions in 1000mL of water for 2 hours. Then, heat the induction cooker at 1300W and boil the liquid. Once the liquid begins to boil, reduce the power to 120W and continue heating for 1.5 hours. Filter the liquid to obtain liquid a. Then, add another 1000mL of water and boil the liquid for 1.5 hours using the same method. Filter the liquid to obtain liquid b. Mix liquid a and liquid b, then heat and concentrate the liquid to 100mL. Finally, refrigerate the liquid for later use.

[0092] 1.5 Animal grouping and intervention

[0093] After one week of acclimatization, mice were randomly divided into four groups of 10 mice each. The four groups were: normal control group, polycystic ovary syndrome fatty liver model group, lipid-regulating and liver-benefiting decoction group (using lipid-regulating and liver-benefiting decoction formula I) (60g / kg / d), positive drug group (metformin tablets 200mg / kg / d by gavage + ethinylestradiol and cyproterone acetate tablets 0.2mg / kg / d), model group and drug treatment group were subcutaneously injected with DHT 2μg / g / d, normal control group was fed standard diet, and the other groups were fed high-fat diet with a fat energy ratio of 40%.

[0094] Six weeks after intervention, mice were fasted for 12 hours, then euthanized by decapitation. Blood was collected, allowed to stand for a period of time, and then centrifuged to separate serum (3,500 rpm, 4°C, 10 min). The serum was stored at -80°C. Liver and ovarian tissues were rapidly dissected from the mice. A small portion of the liver tissue sample was fixed in 4% paraformaldehyde solution and stored at 4°C for morphological observation. The remaining liver and ovarian tissue samples were flash-frozen in liquid nitrogen and stored at -80°C for subsequent assays.

[0095] 2. Results and Analysis

[0096] 2.1 Effects of Tiaozhi Yigan Decoction on Hepatic Triglycerides (TG) in Fatty Liver Mice with Polycystic Ovary Syndrome

[0097] Compared with the normal control group, the TG level in the liver tissue of the model group was significantly increased (p<0.05). After treatment with the lipid-regulating and liver-benefiting decoction, the TG level in the lipid-regulating and liver-benefiting decoction group was significantly decreased (p<0.05). After treatment with the positive control group, the TG level in the positive control group decreased, but the difference was not statistically significant. Figure 9 ).

[0098] 2.2 Effects of Tiaozhi Yigan Decoction on the Histopathological Changes of Liver Tissue in Mice with Fatty Liver Due to Polycystic Ovary Syndrome

[0099] In the normal control group, hepatocytes were morphologically normal, arranged regularly and densely, and evenly distributed, with no obvious fat vacuoles. In the model group, the liver tissue of mice showed structural lesions, including fatty degeneration and ballooning fatty degeneration, hepatocyte damage and necrosis, inflammatory cell infiltration, and a large number of fat vacuoles. The number of fat vacuoles in the lipid-regulating and liver-benefiting decoction group was significantly reduced compared with the normal control group, indicating a significant therapeutic effect. Compared with the model group, the positive drug group showed a reduction in fat vacuoles, and the therapeutic effect of the lipid-regulating and liver-benefiting decoction was better than that of the positive drug group. Figure 10 and Figure 11 ).

[0100] 2.3 Effects of Tiaozhi Yigan Decoction on Ovarian Histopathological Changes in Mice with Fatty Liver Due to Polycystic Ovary Syndrome

[0101] In the control group, the ovarian tissue of mice showed normal granulosa cell layer structure, with visible corpora lutea and follicles at different developmental stages. In the model group, the ovarian tissue of rats exhibited polycystic changes, with reduced or absent corpora lutea, numerous fluid-filled large cystic follicles and atretic follicles, and disordered arrangement and reduced number of granulosa cell layers. Compared to the model group, the ovarian tissue of mice in the lipid-regulating and liver-benefiting decoction group showed fewer cystic and atretic follicles, and the granulosa cell layer structure tended towards normal. The positive control group also showed some improvement. Figure 12 ).

[0102] 2.4 Effects of Tiaozhi Yigan Decoction on MDA Content in Liver Tissue of Mice with Fatty Liver Due to Polycystic Ovary Syndrome

[0103] Compared with the normal control group, the malondialdehyde (MDA) level in the liver tissue of mice in the polycystic ovary syndrome fatty liver model group was significantly increased (p<0.01). After treatment with Tiaozhi Yigan Decoction, the MDA level in both the Tiaozhi Yigan Decoction group and the positive drug group was significantly decreased (p<0.05), and the treatment effect of the Tiaozhi Yigan Decoction group was better than that of the positive drug group. Figure 13 ).

[0104] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A traditional Chinese medicine composition for the prevention or treatment of fatty liver disease, characterized in that: The traditional Chinese medicine composition, by weight, is made from the following raw materials: 5-30 parts of Atractylodes lancea, 5-30 parts of processed Cyperus rotundus, 5-30 parts of Cuscuta chinensis, 5-30 parts of Rubus idaeus, 5-30 parts of Lycium barbarum, 5-30 parts of Viscum album, 5-30 parts of stir-fried Dipsacus asper, 5-30 parts of salt-processed Eucommia ulmoides, 5-30 parts of Salvia miltiorrhiza, and 5-30 parts of Ligusticum chuanxiong. The fatty liver disease is simple fatty liver or polycystic ovary syndrome complicated with fatty liver.

2. The traditional Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition, by weight, is made from the following raw materials: 5-15 parts of Atractylodes lancea, 5-15 parts of processed Cyperus rotundus, 10-20 parts of Cuscuta chinensis, 10-20 parts of Rubus idaeus, 5-15 parts of Lycium barbarum, 10-20 parts of Viscum album, 10-20 parts of stir-fried Dipsacus asper, 5-15 parts of salt-processed Eucommia ulmoides, 10-20 parts of Salvia miltiorrhiza, and 5-15 parts of Ligusticum chuanxiong.

3. The traditional Chinese medicine composition according to claim 2, characterized in that: The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 12 parts of Atractylodes lancea, 10 parts of processed Cyperus rotundus, 15 parts of Cuscuta chinensis, 15 parts of Rubus idaeus, 10 parts of Lycium barbarum, 15 parts of Viscum album, 15 parts of stir-fried Dipsacus asper, 10 parts of salt-processed Eucommia ulmoides, 15 parts of Salvia miltiorrhiza, and 10 parts of Ligusticum chuanxiong.

4. The method for preparing the traditional Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The preparation method includes the following steps: crushing Salvia miltiorrhiza, Cuscuta chinensis, Rubus idaeus, Loranthus parasiticus, stir-fried Dipsacus asper, Atractylodes lancea, processed Cyperus rotundus, Lycium barbarum, salt-processed Eucommia ulmoides, and Ligusticum chuanxiong into powder, adding water to decoct, mixing the filtrate, and concentrating to obtain a concentrated solution.

5. The preparation method according to claim 4, characterized in that, The decoction is prepared twice. After the first decoction is prepared for 1-3 hours, the liquid is filtered to obtain the first filtrate. The residue is then added to water and decocted for another 1-3 hours, and filtered to obtain the second filtrate. The first and second filtrates are combined.

6. The preparation method according to claim 5, characterized in that: The ratio of ingredients to liquid was 1:4 for both the first and second decoctions, and the decoction time was 1.5 hours for both. Distilled water was used for both the first and second decoctions.

7. The preparation method according to claim 4, characterized in that: The concentration is a vacuum concentration.

8. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3, or the traditional Chinese medicine composition prepared by the preparation method according to any one of claims 4 to 7, in the preparation of a medicament for the prevention or treatment of fatty liver disease, characterized in that: The fatty liver disease mentioned refers to simple fatty liver or fatty liver complicated by polycystic ovary syndrome.

Images