Traditional Chinese medicine composition and application thereof in medicine for improving non-alcoholic fatty liver

Abstract

The invention discloses a traditional Chinese medicine composition and application thereof in medicine for improving non-alcoholic fatty liver, and the traditional Chinese medicine composition comprises pulvis fellis suis, pericarpium citri reticulatae, hawthorn and clostridium butyricum. On one hand, potential targets and pathways of drug diseases are determined in network pharmacological analysis, and on the other hand, in an in-vivo experiment, the drug provided by the invention improves obesity and impaired glucose tolerance caused by HFD, reduces the blood fat level, alleviates inflammation, protects the liver, alleviates liver fat vacuole deposition and fatty degeneration, and has a good application prospect. It is proved that the bile acid-hawthorn and pericarpium citri reticulatae-clostridium butyricum combination has prevention and treatment and protection effects on the HFD-induced non-alcoholic 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 and its application in drugs for improving non-alcoholic fatty liver disease. Background Technology

[0002] Nonalcoholic fatty liver disease (NAFLD) is characterized by the accumulation of large amounts of ectopic triglycerides in the liver in the absence of other liver diseases or excessive alcohol consumption. NAFLD is the most common liver disease, affecting 20%-30% of adults and over 80% of obese individuals worldwide. NAFLD can progress to nonalcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma. As a component of metabolic syndrome, NAFLD is closely associated with insulin resistance, type 2 diabetes, coronary heart disease, and atherosclerosis. Furthermore, NAFLD is a risk factor for hepatocellular carcinoma. To date, the underlying molecular mechanisms of NAFLD pathogenesis remain largely unclear. Treatment for NAFLD mainly includes a healthy, balanced diet, physical exercise, and some synthetic drugs containing metformin, orlistat, and vitamin E. However, these regimens have certain side effects and cannot meet the needs. Therefore, there is an urgent need to develop novel drugs for the prevention and treatment of NAFLD.

[0003] NAFLD has multi-target characteristics, but currently, most commonly used acidic polysaccharides are derived from single Chinese herbal medicines, such as Cornus officinalis (CN113999328B), Eucommia ulmoides (CN113717295B), Lonicera japonica (CN115304687A), Arctium lappa root (CN111084784A), and Tremella fuciformis (CN114073711A). Their efficacy is unsatisfactory, necessitating the combination of acidic polysaccharides from Chinese herbal medicines with specific activities for NAFLD treatment. Numerous methods for preventing and treating fatty liver have been reported in the literature, including various Chinese herbal formulas such as hawthorn, turmeric, Polygonum multiflorum, Atractylodes lancea, Alisma plantago-aquatica, Poria cocos, Paeonia lactiflora, Angelica sinensis, and Gentiana scabra. However, the success rate of these treatments in actual clinical use is not high, and most Chinese herbal formulas are too expensive for practical clinical application. Summary of the Invention

[0004] The problem solved by this invention is to find and provide a simple traditional Chinese medicine composition that can effectively improve non-alcoholic fatty liver disease based on network pharmacology analysis. This traditional Chinese medicine composition has the advantages of simple formulation, specific efficacy, precise regulation, and significant effect.

[0005] This invention addresses the above problems through a combination of methods, and the technical solution is as follows:

[0006] A traditional Chinese medicine composition comprising pig bile powder, dried tangerine peel, hawthorn, and Clostridium butyricum.

[0007] The technical principle of this invention: The drug combination involved in this invention has the advantages of definite efficacy, high safety, abundant target sites, few side effects, and low price. Bile acids are an important component of bile, and are a series of sterols produced during cholesterol metabolism in animals. Bile acids are mainly present in the enterohepatic circulation system and have multiple biological activities through recycling. In the enterohepatic circulation, bile acids emulsify fats and activate lipase activity to hydrolyze fats, thereby forming chylomicrons with fatty acids, allowing fatty acids to be absorbed through the membrane, thus promoting the efficient digestion and absorption of fats. In addition, bile acids returning to the liver can stimulate bile secretion, clear the bile ducts, and improve metabolism.

[0008] Hawthorn can increase fatty acid oxidation and lower blood lipid levels, helping to reduce fat accumulation in the liver. Its antioxidant components can scavenge free radicals, reduce oxidative stress, and thus reduce hepatocyte damage and inflammation. By promoting normal insulin function, it helps improve glucose metabolism and alleviate NAFLD symptoms. Dried tangerine peel also regulates lipid metabolism, has anti-inflammatory effects, and promotes bile secretion. Combined use with bile acids can promote enterohepatic circulation and improve metabolism. Clostridium butyricum has good resistance to high temperatures, acids, and bile salts, and exhibits some resistance to the high temperature and pressure during feed processing and the digestive tract environment. In vitro experiments have shown that Clostridium butyricum can secrete digestive enzymes such as amylase and protease, increasing the activity of amylase, lipase, and protease in the intestinal chyme of piglets to aid digestion and improve the apparent digestibility of dry matter, crude protein, and calcium and phosphorus in feed. Animal experiments have shown that adding Clostridium butyricum can alleviate the decrease in amylase, lipase, and protease activity in intestinal chyme caused by Escherichia coli infection. In July 2009, my country's Ministry of Agriculture and Rural Affairs approved the use of Clostridium butyricum as a microbial additive (Announcement No. 1231). In 2013, based on the "Catalogue of Feed Additive Varieties (2008)," Clostridium butyricum was officially included in the new catalogue of feed additive products.

[0009] Furthermore, the raw material, pig bile powder, was purchased from Hangzhou Baoji Biotechnology, hawthorn and dried tangerine peel were purchased from a pharmacy, and Clostridium butyricum was provided by Hubei Languzhong Microbial Technology Co., Ltd.

[0010] Furthermore, in the aforementioned traditional Chinese medicine composition, the mass ratio of pig bile powder, dried tangerine peel, and hawthorn is 2:1:1.

[0011] Furthermore, the porcine bile powder contains ≥16.0% porcine deoxycholic acid.

[0012] Furthermore, the hawthorn and dried tangerine peel are pulverized and sieved through a 200-mesh sieve for later use.

[0013] Furthermore, the *Clostridium butyricum* was prepared at a concentration of 1×10⁻⁶. 8 CFU / each / day for drinking water use.

[0014] This invention also provides the application of a traditional Chinese medicine composition in a drug for improving non-alcoholic fatty liver disease in mice.

[0015] Furthermore, the application of the aforementioned traditional Chinese medicine composition in improving non-alcoholic fatty liver disease in mice involved using deoxycholic acid, the main component of porcine bile powder, along with hawthorn and tangerine peel, to conduct network pharmacological analysis on non-alcoholic fatty liver disease and hyperlipidemia. The analysis revealed that this drug combination can regulate lipid metabolism through target pathways such as TNF-α, PPAR family, TP53, and STAT3, thereby preventing and treating non-alcoholic fatty liver disease.

[0016] Furthermore, the application of the aforementioned traditional Chinese medicine composition in improving non-alcoholic fatty liver disease in mice, wherein the combination of pig bile powder, hawthorn, tangerine peel, and Clostridium butyricum inhibits obesity and impaired glucose tolerance in mice with HFD-induced non-alcoholic fatty liver disease.

[0017] Furthermore, the application of the aforementioned traditional Chinese medicine composition in improving non-alcoholic fatty liver disease in mice, wherein the combination of pig bile powder, hawthorn, tangerine peel, and Clostridium butyricum inhibits the increase in blood lipids in mice with HFD-induced non-alcoholic fatty liver disease.

[0018] Furthermore, the application of the aforementioned traditional Chinese medicine composition in the treatment of non-alcoholic fatty liver disease in mice, wherein the combination of pig bile powder, hawthorn, tangerine peel, and Clostridium butyricum improves hepatic steatosis and degeneration.

[0019] The traditional Chinese medicine composition of this invention has been demonstrated through network pharmacology analysis to have the effect of preventing and treating non-alcoholic fatty liver disease. 2084 disease targets were obtained from three databases: Genecards, Disgenet, and OMIM. These targets were mapped with 254 potential targets of the compound's effective components, resulting in 213 potential targets with common effects. Protein-protein interactions (PPIs) were explored using the String database for these 213 targets, identifying tumor necrosis factor-α (TNF-α), peroxisome proliferator-activated receptor (PPARG), and signal transduction and transcription activator (STAT3) as core targets. KEGG and GO enrichment analyses were performed on the 213 overlapping targets, revealing key pathways such as HIF-1 signaling, lipid and atherosclerosis, fluid shear stress and atherosclerosis, as well as major biological processes such as inflammatory response, positive regulation of nitric oxide biosynthesis, and positive regulation of gene expression, and molecular functions such as protein binding, RNA polymerase transcription factor activity, enzyme activity, and protein binding.

[0020] This invention uses male C57BL / 6J mice to establish a non-alcoholic fatty liver disease animal model, and conducts in vivo studies on the above-mentioned traditional Chinese medicine composition. The results are as follows:

[0021] (1) After the model mice were given the combination of drugs of the present invention, their body weight and liver index were significantly reduced, and their glucose tolerance was improved.

[0022] (2) In mice with non-alcoholic fatty liver disease, administration of the herbal combination of the present invention significantly reduced serum total cholesterol (TC) and triglycerides (TG). In model mice, administration of the herbal combination of the present invention significantly reduced serum cholesterol and triglycerides, decreased low-density lipoprotein (LDL) levels, increased high-density lipoprotein (HDL) levels, and improved liver function.

[0023] (3) HE staining of liver sections revealed that the drug of the present invention has a positive effect on maintaining the structural integrity of the liver, effectively improves liver damage caused by non-alcoholic fatty liver, significantly improves the tightness of hepatocyte arrangement, reduces the number and size of fat vacuoles, improves cells and structure and reduces the infiltration of inflammatory cells. Attached Figure Description

[0024] Figure 1 This is a network pharmacological analysis diagram of bile acids and traditional Chinese medicine. Figure 1 a is a Venn diagram of mouse NAFLD targets from three databases: Genecards, Disgenet, and OMIM. Figure 1 b is a Venn diagram of targets for hyperlipidemia in mice from the Genecards, Disgenet, and OMIM databases. Figure 1 c is a Venn diagram of common targets for NAFLD and hyperlipidemia in mice from the Genecards, Disgenet, and OMIM databases. Figure 1 d is the Venn diagram of the intersection of bile acids, hawthorn, tangerine peel and mouse NAFLD and hyperlipidemia; Figure 1 e and f are PPT diagrams showing how 213 drug-disease targets were explored using the String database to identify protein-protein interactions (PPIs). Figure 1 g is a KEGG enrichment analysis plot; Figure 1 h is the GO enrichment analysis plot.

[0025] Figure 2a , Figure 2b , Figure 2c The diagrams show the changes in body weight, liver index, and oral glucose tolerance test results for each group of mice. Figure 2a This invention indicates that the herbal combination can alleviate weight gain caused by a high-risk diet (HFD). Figure 2b This invention demonstrates that the herbal combination can improve the elevated liver index in mice induced by HFD. Figure 2c This indicates that the herbal combination of the present invention can repair glucose tolerance impairment in mice to a certain extent.

[0026] Figure 3 This is a graph showing blood lipid levels 10 weeks after experimental drug administration. Figure 3 a) The herbal combination of the present invention can inhibit the increase of serum triglycerides in HFD-induced NAFLD mice; Figure 3 b is that the herbal combination of the present invention can inhibit the increase of serum cholesterol in HFD-induced NAFLD mice; Figure 3 c is that the combination of traditional Chinese medicines in this invention can inhibit the decrease of serum high-density lipoprotein in HFD-induced NAFLD mice; Figure 3 d indicates that the herbal combination of the present invention can inhibit the increase of serum low-density lipoprotein in HFD-induced NAFLD mice.

[0027] Figure 4 This is an image of a mouse liver tissue section stained with hematoxylin and eosin (HE). Detailed Implementation

[0028] The present invention can be better understood from the following embodiments and accompanying drawings. The specific addition ratios and their results described in the embodiments are for illustrative purposes only and are not intended to limit the scope of protection of the present invention. After reading the contents of this invention, those skilled in the art can make various alterations or modifications to the present invention, and these equivalent changes and modifications also fall within the scope defined by the claims of the present invention.

[0029] Example 1

[0030] Network pharmacology was used to analyze and predict the effects of combined drugs on non-alcoholic fatty liver disease (NAFLD) in mice.

[0031] 1. Identification of TCM component targets: Using TCMSP, dried tangerine peel was collected, screened for OB≥30% and DL≥0.18, yielding 5 components and 63 targets. Hawthorn was screened using the BATMAN 2.0 database, selecting only known targets, resulting in 28 components and 354 targets. The structure and SMILE numbers of "Hyodeoxycholic acid" were collected using PubChem, followed by target prediction using Swiss targets prediction, yielding 100 targets. Uniprot annotation of protein names resulted in 33 components and 467 deduplicated targets. A Cytoscape network was constructed, and the network topology was calculated, showing 503 nodes and 725 edges.

[0032] 2. Disease Target Acquisition: The databases used for disease identification were Genecards, Disgenet, and OMIM, with searches conducted using the keywords "non-alcoholic fatty liver disease" and "hyperlipidemia." Screening criteria: For "non-alcoholic fatty liver disease," Genecards used a Relevance score ≥ 1, yielding 578 targets. Disgenet used a score_gda ≥ 0.1, yielding 70 targets. OMIM only selected approved targets, yielding 154 targets. For "hyperlipidemia," Genecards used a Relevance score ≥ 5, yielding 1852 targets. Disgenet used a score_gda ≥ 0.1, yielding 140 targets. OMIM only selected approved targets, yielding 2 targets. Finally, the union of the targets for these two diseases was calculated, resulting in 2297 targets.

[0033] Target intersection determination: The intersection of the targets of traditional Chinese medicine components and the targets of diseases was determined, and a total of 213 common targets were obtained.

[0034] PPI analysis: Protein-protein interactions (PPIs) were explored for these 213 targets using the String database, with a confidence interval of 0.9. Visual analysis was then performed using Cytoscape software, adjusting the size and staining based on the degree value; darker colors corresponded to larger circles with higher values.

[0035] GO and KEGG enrichment analysis: KEGG and GO enrichment analysis was performed on 213 intersecting target sites, see [link to relevant documentation]. Figure 1 As shown, KEGG pathway enrichment analysis revealed that the HIF-1 signaling pathway, lipid and atherosclerosis, and fluid shear stress and atherosclerosis were the main pathways. GO enrichment analysis mainly involved biological processes (BP) such as inflammatory response, positive regulation of nitric oxide biosynthesis, and positive regulation of gene expression. The main cellular components (CC) involved were cytoplasm, membrane rafts, plasma membrane components, and plasma membranes. The main molecular functions (MF) involved were protein binding, RNA polymerase transcription factor activity, enzyme activity, and protein binding.

[0036] Example 2

[0037] To prepare a medicated feed for mice, pig bile powder, hawthorn, dried tangerine peel, and Clostridium butyricum were added to a high-fat diet for mice in specific proportions. The pig bile powder was mixed with hawthorn and dried tangerine peel in a mass ratio of 2:1:1, and 1×10⁻⁶ Clostridium butyricum was added to the feed. 8 CFU / each / day for drinking water use.

[0038] Example 3

[0039] Animal experiments demonstrating the in vivo therapeutic effect of the traditional Chinese medicine composition described in this invention on non-alcoholic fatty liver disease.

[0040] 1. Test drug

[0041] Nonalcoholic fatty liver disease was tested using the drug combination method described in Example 2.

[0042] 2. Laboratory animals

[0043] C57BL / 6J mice, male, provided by Changsha Tianqin Biotechnology Co., Ltd.

[0044] 3. Experimental Methods

[0045] 3.1 The high-fat diet for mice was purchased from Changsha Tianqin Biotechnology Co., Ltd. The formula consisted of 18% lard, 20% white sugar, 3% egg yolk, and the remainder was the basic feed.

[0046] 3.2 Grouped administration

[0047] After two weeks of acclimatization, mice were used to establish a bile acid drug model. The animals were randomly divided into a control group, a model group, and the bile acid drug group of this invention. Drug administration began in the seventh week of modeling and continued until the sixteenth week, for a total of 10 weeks.

[0048] 3.3 Specimen Collection

[0049] In week 16, a glucose tolerance test was conducted. At the end of week 16, the animals were fasted for 12 hours, weighed, and blood was collected from the eyeballs of mice in each group. Immediately after blood collection, all mice were euthanized by cervical dislocation. The blood was centrifuged at 3500 rpm for 15 minutes at 4 degrees Celsius. The liver tissue was collected, fixed, and cryopreserved. Half of the mouse liver was preserved in 4% paraformaldehyde solution, dehydrated, and embedded in paraffin to form a paraffin block. Subsequently, the liver tissue was sectioned to a thickness of 4 μm and examined by routine H&E staining.

[0050] 3.4 Indicator Testing

[0051] The levels of TC, TG, HDL, and LDL in mouse blood biochemical parameters were detected using a kit in serum samples, and liver sections were prepared for HE staining.

[0052] HE staining:

[0053] ① Dehydration: The liver tissue blocks fixed with 4% paraformaldehyde were trimmed with a scalpel, rinsed with running water overnight, and then dehydrated with a gradient of alcohol. The alcohol concentration and time required for dehydration were: 70% alcohol (40 min) → 85% alcohol (40 min) → 90% alcohol (40 min) → 95% alcohol (20 min) → 95% alcohol (20 min) → 100% alcohol (50 min) → 100% alcohol (50 min).

[0054] ②Clearing: The tissue block was treated with xylene. The xylene time required for clearing was: xylene I (10 min) → xylene II (10 min);

[0055] ③ Paraffin infiltration: Immerse the tissue block in melted paraffin. The required infiltration time is: Paraffin I (60 min) → Paraffin II (60 min).

[0056] ④ Embedding: Gently place the fully paraffin-impregnated tissue block into the paraffin embedding cassette, and adjust the position of the tissue block so that it is located in the center;

[0057] ⑤ Sectioning: Cut the completely cooled tissue paraffin block into 4µm thick sections using a microtome;

[0058] ⑥ Mounting: First, apply an appropriate amount of protein glycerin to the glass slide, then attach the cut slices to the glass slide and dry;

[0059] ⑦ HE staining: Reagents and times required for staining: Xylene I (10 min) → Xylene II (5 min) → 100% alcohol (3 min) → 90% alcohol (3 min) → 85% alcohol (3 min) → 70% alcohol (3 min) → distilled water (3 min) → hematoxylin staining solution (2 min) → rinse with running water to reverse blue (10 min) → eosin staining solution (30 s) → 95% alcohol (4 min) → 95% alcohol (4 min) → 100% alcohol (4 min) → Xylene I (10 min) → Xylene II (5 min);

[0060] ⑧ Covering: Add a drop of neutral resin and carefully cover with a coverslip;

[0061] ⑨ Microscopic examination: Place the prepared slides under a light microscope for examination, take pictures under a 40x microscope, and observe histological and pathological changes.

[0062] 4. Results

[0063] The effects of the traditional Chinese medicine composition of this invention on body weight, liver index, and glucose tolerance in mice with non-alcoholic fatty liver disease (NAFLD): This composition significantly improves obesity, excessive liver index, and impaired glucose tolerance in mice. Studies show that the traditional Chinese medicine composition of this invention can effectively control body weight gain in NAFLD mice, such as... Figure 2a As shown, after drug administration, the body weight of mice in the model group was higher than that in the control group and the drug-treated group, and there was a significant difference in body weight between the model group and the other groups (P < 0.05). Figure 2b As shown, the liver index of mice in the model group was higher than that in the control group and the drug-treated group, and the differences were statistically significant (P < 0.05). Figure 2c As shown, after oral administration of glucose, the blood glucose levels in the control group initially increased and then gradually returned to normal within 120 minutes, while the blood glucose levels in the model group remained high and were still at a high level at 120 minutes, significantly higher than the control group (P < 0.05). The blood glucose levels in the H1 group recovered better than those in the model group after 120 minutes. This indicates that the traditional Chinese medicine combination of the present invention can repair impaired glucose tolerance.

[0064] The drug of this invention can inhibit the trend of elevated blood lipids in mice caused by non-alcoholic fatty liver disease induced by a high-fat diet.

[0065] A high-fat diet is one of the main factors leading to NAFLD. Long-term intake of high-fat foods can cause an imbalance in lipid metabolism, resulting in abnormally high blood lipids and fat accumulation in the liver, eventually developing into fatty liver disease. The drug of this invention effectively reduces serum cholesterol, triglycerides, and other blood lipid indicators by regulating lipid metabolism, thereby reducing fat accumulation in the liver and combating NAFLD caused by a high-fat diet.

[0066] Hyperlipidemia is the basis for the development of non-alcoholic fatty liver disease, and elevated blood lipids are one of the main manifestations of hyperlipidemia. The results are as follows: Figure 3 As shown, compared with the normal group, the model group animals showed significantly elevated serum triglycerides and cholesterol levels. After 10 weeks of continuous administration, compared with the control group, as... Figure 3 a. Triglyceride (TG) levels were 1.4 times higher than in the control group; Figure 3 b, The model group showed a 2.7-fold increase in total cholesterol (TC); Figure 3 c. Low-density lipoprotein (LDL) was 4 times that of the control group; Figure 3 d. High-density lipoprotein (HDL) was 0.69 times that of the blank group. There was no significant difference between the drug group and the blank group (P>0.05), which proves that the herbal composition of the present invention can significantly reduce serum triglyceride (TG), cholesterol (TC) and serum low-density lipoprotein (LDL) levels. The drug group can inhibit the decrease in serum HDL level in mice caused by HFD, and the drug group recovers to normal level.

[0067] In the control group, the liver tissue of mice was intact, with hepatocytes arranged neatly, clear nuclei, and no pathological changes such as fatty degeneration or swelling. In the model group, hepatocytes were disordered, with nuclei pushed to one side, and cytoplasmic fat vacuoles of varying sizes, indicating inflammatory cell infiltration. In the group containing the herbal composition of this invention, the hepatocytes of mice were neatly arranged, with normal nuclei, a small number of fat vacuoles, and a small number of inflammatory cell infiltrations. The results are shown in the figure. Figure 4 ,Depend on Figure 4 It is known that the herbal combination of the present invention can alleviate fat deposition and inflammatory cell infiltration.

[0068] 5. Conclusion

[0069] Studies show that the traditional Chinese medicine combination of this invention can effectively control excessive weight gain in NAFLD mice and contribute to weight reduction by regulating lipid metabolism and glucose tolerance. The liver index (the ratio of liver weight to body weight) is an important indicator for assessing the degree of hepatic fat accumulation, and the drug of this invention can significantly reduce the liver index in NAFLD mice, indicating that it can effectively reduce hepatic fat accumulation and hepatomegaly. This may be attributed to the synergistic effect of specific components in the composition, which may promote fatty acid oxidation, inhibit lipid synthesis, and improve the liver's antioxidant capacity, thereby reducing fat accumulation in the liver. Impaired glucose tolerance is one of the hallmarks of metabolic abnormalities in non-alcoholic fatty liver disease. The drug of this invention effectively improves glucose tolerance in non-alcoholic fatty liver mice, indicating that the drug improves glucose metabolism by promoting efficient glucose utilization. This effect may be related to the active ingredients in the traditional Chinese medicine composition acting directly on pancreatic β cells or indirectly affecting glucose metabolism by regulating the gut microbiota.

[0070] This invention's drug, through comprehensive regulation of lipid metabolism, exhibits significant effects in combating dyslipidemia and NAFLD caused by a high-fat diet. It reduces serum triglyceride (TG) and total cholesterol (TC) levels, significantly lowers low-density lipoprotein (LDL-C) levels, and upregulates high-density lipoprotein (HDL-C) levels after administration. Furthermore, this invention's drug has a positive effect on maintaining liver structural integrity, effectively improving liver damage caused by non-alcoholic fatty liver disease, significantly improving the density of hepatocyte arrangement, reducing the number and size of fat vacuoles, improving cell structure, and reducing inflammatory cell infiltration.

[0071] The drug combination of this invention achieves its therapeutic effect mainly through the following pathways:

[0072] Regulation of weight, glucose tolerance, and blood lipid levels: The drug combination of this invention reduces weight, repairs impaired glucose tolerance, and lowers serum triglyceride (TG) and total cholesterol (TC) levels. This may be achieved by influencing fatty acid synthesis and breakdown, reducing endogenous lipid production, and promoting the oxidative breakdown of fatty acids.

[0073] Improves lipoprotein composition: Reduces levels of harmful low-density lipoprotein (LDL) while increasing levels of beneficial high-density lipoprotein (HDL).

[0074] Anti-inflammatory: It reduces hepatic steatosis and degeneration, as well as inflammatory cell infiltration, and improves hepatocellular damage, thereby protecting the liver.

Claims

1. A traditional Chinese medicine composition, characterized in that, Pig gall powder, dried tangerine or orange peel, hawthorn, Clostridium butyricum.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The mass ratio of pig gall powder, dried tangerine or orange peel and hawthorn is 2:1:

1.

3. The traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The content of hyodeoxycholic acid in the pig gall powder is greater than or equal to 16.0%.

4. The traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The hawthorn and dried tangerine or orange peel are crushed and sieved to 200 meshes.

5. The traditional Chinese medicine composition according to claim 1, characterized in that, The Clostridium butyricum is used at 1 x 10 8 CFU / animal / day in drinking water.

6. The application of the traditional Chinese medicine composition according to any one of claims 1-5 in improving non-alcoholic fatty liver disease in mice. 7.The application of the traditional Chinese medicine composition of claim 6 in improving non-alcoholic fatty liver disease in mice, characterized in that, The network pharmacology analysis of the main component of pig gall powder, hawthorn and dried tangerine or orange peel is used to find that the drug combination can regulate lipid metabolism through the target point pathways of TNF-alpha, PPAR family, TP53 and STAT3 to prevent and treat non-alcoholic fatty liver disease. 8.The application of the traditional Chinese medicine composition of claim 6 in improving non-alcoholic fatty liver disease in mice, characterized in that, The pig gall powder, hawthorn, dried tangerine or orange peel and Clostridium butyricum combination inhibits obesity and impaired glucose tolerance of non-alcoholic fatty liver mice induced by HFD. 9.The application of the traditional Chinese medicine composition of claim 6 in improving non-alcoholic fatty liver disease in mice, characterized in that, The pig gall powder, hawthorn, dried tangerine or orange peel and Clostridium butyricum combination inhibits the increase of blood lipids of non-alcoholic fatty liver mice induced by HFD. 10.The application of the traditional Chinese medicine composition of claim 6 in improving non-alcoholic fatty liver disease in mice, characterized in that, The pig gall powder, hawthorn, dried tangerine or orange peel and Clostridium butyricum combination improves liver fat deposition and degeneration.

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