Traditional Chinese medicine composition for improving glucose and lipid metabolism, body fat level and non-alcoholic fatty liver disease and its use

Abstract

The present invention belongs to the technical field of traditional Chinese medicine, and specifically relates to a traditional Chinese medicine composition and use for improving glucose and lipid metabolism, body fat levels and non-alcoholic fatty liver disease. The technical problem to be solved by the present invention is to provide a new option for treating obesity, glucose and lipid metabolism disorders and non-alcoholic fatty liver disease. The technical solution of the present invention is a traditional Chinese medicine composition for improving glucose and lipid metabolism, body fat levels and non-alcoholic fatty liver disease, comprising the following ingredients: ginseng, malt, poria, buzia leaves, prunella vulgaris and sophora japonica flowers. The traditional Chinese medicine composition of the present invention has the effects of soothing the liver, regulating qi and strengthening the spleen, clearing the liver, purging fire and dispersing stagnation, and is used for weight loss, lipid reduction, blood sugar reduction, improving body fat rate, treating liver steatosis and inflammation, and improving liver fibrosis. It has a significant effect, and can also prevent the occurrence of cardiovascular and cerebrovascular diseases induced by glucose and lipid metabolism disorders through lipid reduction, anti-inflammatory, anti-oxidation and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and specifically relates to a traditional Chinese medicine composition and use thereof for improving glucose and lipid metabolism, body fat levels and non-alcoholic fatty liver disease. Background Art

[0002] Non-alcoholic fatty liver disease (NAFLD) is a metabolic stress-induced liver injury closely related to insulin resistance and genetic susceptibility. The disease spectrum includes non-alcoholic simple fatty liver (NAFL), non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma. It has become the leading chronic liver disease in my country. The pathogenesis of NAFLD is not fully understood, but it is currently believed to stem from an overload of the liver's ability to process primary energy metabolic substrates, carbohydrates, and fatty acids, leading to the accumulation of toxic lipids. Nearly 60% of patients with simple hepatic steatosis may progress to non-alcoholic steatohepatitis, characterized by inflammation and hepatocellular damage. 41% of patients with NAFLD also develop fibrosis. Of these, 22% of patients with advanced fibrosis may progress to cirrhosis, and 2-3% of patients with cirrhosis may develop hepatocellular carcinoma (HCC) within three years. This chronic liver disease has been dubbed a "silent killer." Patients with NAFLD often suffer from comorbidities such as obesity (51.3%), hyperlipidemia (69.2%), hypertension (39.3%), and type 2 diabetes mellitus (T2DM) (22.5%). Furthermore, NAFLD significantly increases the risk of cardiovascular disease (CVD), with atherosclerosis having the highest prevalence (55.5%). The harmful effects of NAFLD on the body not only lead to liver disease, disability, and death, but are also closely associated with a high incidence of metabolic syndrome, type 2 diabetes, arteriosclerotic cardiovascular disease, and colorectal cancer, leading to increased morbidity and mortality from diabetes and cardiovascular disease. Therefore, strengthening the prevention of NAFLD and preventing its further development is of great value and significance.

[0003] The Traditional Chinese Medicine (TCM) concept of "preventive treatment" has advantages in preventing and treating such complex diseases. Therefore, there is an urgent need to find foods, health products, and medicines that are effective and safe in preventing the occurrence and development of NAFLD. The TCM names for NAFLD are based on analysis of its symptoms, etiology, and pathogenesis, and are generally classified as "liver problems," "flank pain," and "accumulation." Spleen deficiency and dysfunctional transportation and transformation are the root of the pathogenesis. The spleen is the "foundation of acquired constitution," responsible for transportation and transformation, distributing the essence of water and grain, and is the source of qi and blood production. In modern society, work and life are stressful, leading to emotional imbalance, liver qi stagnation, liver qi invading the spleen, the spleen failing to function properly, and abnormal distribution of essence, which in turn transforms into water, dampness, and phlegm. Turbid pathogens accumulate in the middle burner for a long time, becoming stagnant and turning into heat, resulting in internal dampness and heat. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a new option for improving glucose and lipid metabolism, body fat levels and non-alcoholic fatty liver disease.

[0005] The technical solution of the present invention is a traditional Chinese medicine composition for improving sugar and lipid metabolism, body fat level and non-alcoholic fatty liver disease, comprising the following ingredients: ginseng, malt, poria, bu zai leaves, prunella vulgaris and sophora japonica flowers. Traditional Chinese medicine believes that the location of non-alcoholic fatty liver disease is in the liver, which is closely related to the spleen and stomach. The main causes of the disease are improper diet, emotional disorders, etc., which result in liver failure to regulate qi and spleen failure to function properly, internal accumulation of dampness and heat, and obstruction of the liver meridians. Treatment should be aimed at soothing the liver, regulating qi and strengthening the spleen, clearing the liver, purging fire and dispersing nodules. In the formula, malt soothes the liver and regulates qi, strengthens the spleen and harmonizes the stomach; bu zai leaves, prunella vulgaris and sophora japonica flowers are used together to clear the liver, purge fire, and dissipate nodules and reduce swelling; ginseng replenishes qi and strengthens the spleen, and poria strengthens the spleen and eliminates dampness. The combination of the two medicines enhances the spleen and eliminates dampness. The whole formula is reasonably combined, and has significant effects of soothing the liver, regulating qi and strengthening the spleen, clearing the liver, purging fire and dispersing nodules.

[0006] Furthermore, the weight proportions of the ingredients are as follows: 2-30 parts of ginseng, 1-30 parts of malt, 3-40 parts of Poria cocos, 3-50 parts of Buza leaves, 3-30 parts of Prunella vulgaris and 2-30 parts of Sophora japonica flowers.

[0007] Preferably, the weight proportion of the ingredients is as follows: 3 parts of ginseng, 6 parts of malt, 6 parts of Poria cocos, 7.5 parts of Buza leaves, 4.5 parts of Prunella vulgaris and 3 parts of Sophora japonica flowers.

[0008] The present invention also provides a preparation method of the traditional Chinese medicine composition, comprising the following steps: preparing the following raw materials in parts by weight: 2 to 30 parts of ginseng, 1 to 30 parts of malt, 3 to 40 parts of poria cocos, 3 to 50 parts of buzia leaves, 3 to 30 parts of selfheal and 2 to 30 parts of sophora japonica flowers; adding water, heating and refluxing extraction for 2 times, filtering, combining the two extracts, concentrating and drying.

[0009] Furthermore, during the extraction process, the mass-to-volume ratio of the material to the liquid (g / mL) was 1:3 to 1:20.

[0010] Preferably, during the first extraction, the mass-to-volume ratio of the material to the liquid (g / mL) is 1:10.

[0011] Preferably, during the first extraction, the mass-to-volume ratio of the material to the liquid (g / mL) is 1:8.

[0012] Specifically, each extraction time is 1 to 3 hours.

[0013] Preferably, each extraction time is 1 hour.

[0014] The present invention also provides a traditional Chinese medicine composition obtained by the above preparation method.

[0015] The present invention also provides use of the above-mentioned traditional Chinese medicine composition in preparing a medicine for treating or preventing non-alcoholic fatty liver disease.

[0016] Among them, the application is the use of the above-mentioned traditional Chinese medicine composition in the preparation of drugs for controlling body weight and food intake, reducing body fat percentage, improving glucose intolerance and insulin resistance, improving fatty liver, improving blood lipids, lowering blood sugar, reducing liver damage, improving inflammatory response, improving liver tissue oxidative stress level and / or improving liver damage such as hepatic steatosis, inflammation, fibrosis, etc. caused by a high-fat and high-sugar diet.

[0017] Furthermore, the inflammatory factors involved in the inflammatory response are TNF-α, IL-1β, MCP-1 and IL-10.

[0018] The present invention also provides a medicine for preventing or treating obesity, glucose and lipid metabolism disorders, and non-alcoholic fatty liver disease, comprising the above-mentioned traditional Chinese medicine composition.

[0019] In particular, the above-mentioned medicine also includes pharmaceutically acceptable adjuvants.

[0020] Further, the auxiliary agent is dextrin, CMC-Na or starch.

[0021] Furthermore, the dosage form of the drug is tablets, injections, capsules, pills or granules.

[0022] Beneficial Effects of the Invention: Based on Sijunzi Decoction, the present invention continuously optimizes the formula based on clinical practice. The formula comprises six Chinese medicinal herbs with both medicinal and edible properties: ginseng, malt, poria cocos, buzia leaves, prunella vulgaris, and sophora japonica flowers. The Chinese medicinal composition of the present invention has the effects of soothing the liver, regulating qi, and strengthening the spleen, clearing the liver, purging heat, and dispersing stagnation. It is significantly effective in improving glucose and lipid metabolism, body fat levels, and non-alcoholic fatty liver disease. It can be developed into a drug for regulating glucose and lipid metabolism disorders, promoting weight loss, and preventing and treating non-alcoholic fatty liver disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 . Body weight statistics of each group (n=6, x±s).

[0024] Figure 2 . Fasting blood glucose statistics of each group (n=6, x±s).

[0025] Figure 3 . Appearance of the liver of mice in each group.

[0026] Figure 4 Representative images of H&E-stained liver sections from each group (n=3, x±s).

[0027] Figure 5 Representative images of Oil Red O-stained liver sections from each group (n=3, x±s).

[0028] Figure 6 F4 / 80 immunohistochemical staining of liver sections in each group (n=3, x±s).

[0029] Figure 7 . Picrosirius red staining of liver sections in each group (n=3, x±s). DETAILED DESCRIPTION

[0030] The present invention is based on Sijunzi Decoction and continuously optimizes the prescription in combination with clinical practice to form a fixed prescription, which is composed of 6 Chinese medicinal herbs with medicinal and edible properties, namely ginseng, malt, poria, buzia leaves, selfheal, and sophora japonica flowers. To further confirm its therapeutic efficacy, a NAFLD mouse model consistent with clinical characteristics was established by inducing a high-fat, high-sugar diet based on the pathological characteristics and TCM pathogenesis of NAFLD. The model was tested for lipid regulation (TG, TC, HDL-C, LDL-C) and glucose homeostasis (OGTT, HOMA-IR) indices, liver damage indices (ALT, AST) and oxidative stress indices (MDA, SOD), inflammation (TNF-α, MCP-1, IL-6, IL-1β), and fibrosis-related indices (HA, Col-IV, LN, α-SMA, CoI1α1, Tgf-β1, FN-1). Combined with pathological staining (H&E staining, Oil Red O staining, picrosirius red staining, and immunohistochemistry staining), the pharmacological effects of the compound on improving glucolipid metabolism, body fat levels, and non-alcoholic fatty liver disease were objectively evaluated, providing a scientific basis for its development as a potential drug or health product for improving glucolipid metabolism, body fat levels, and the prevention and treatment of non-alcoholic fatty liver disease.

[0031] Reagents and materials used in the following examples:

[0032] Traditional Chinese medicine (TCM) was purchased from Guangdong Medicinal Materials and Chinese Herbal Medicine Factory, silymarin was purchased from Maboshi Pharmaceutical Factory in Germany, a high-fat diet (60% fat, D12492, Deitz, USA), D-glucose was purchased from Guangdong Guangshi Reagent Technology Co., Ltd., and D-fructose was purchased from Shanghai MacLean Biochemical Technology Co., Ltd. TC, TC, LDL-C, HDL-C, insulin, blood glucose, IL-1β, IL-6, and TNF-α assay kits were purchased from Jiangsu Enzyme Immunoassay Co., Ltd. MDA, SOD, ALT, and AST assay kits were purchased from Nanjing Jiancheng Bioengineering Research Institute Co., Ltd. Six-week-old C57BL / 6J male mice, weighing 18–20 g, were purchased from Zhuhai Baishitong Biotechnology Co., Ltd.

[0033] Example 1 Preparation of the Chinese medicine composition of the present invention

[0034] 3 parts ginseng, 6 parts malt, 7 parts poria cocos, 7.5 parts buzia leaves, 4.5 parts prunella vulgaris, and 3 parts sophora japonica flowers. According to the above prescription, weigh the corresponding weights of the medicinal materials and extract them twice with water, heating under reflux, first with 10 times the amount and then with 8 times the amount, each time for 1 hour. Filter, combine the two extracts, concentrate under reduced pressure, and dry to obtain an extract powder.

[0035] Example 2 Effect Verification of the Chinese Medicine Composition of the Present Invention

[0036] The mice were divided into a blank control group (Con), a model group (Mod), a positive drug silymarin group (Sily, 100 mg / kg), a modified prescription (i.e., the Chinese medicine composition of the present invention) low-dose group (Modified prescription-L, 2.6 g / kg), and a modified prescription high-dose group (Modified prescription-H, 5.2 g / kg) according to their body weight using a random number table method.

[0037] After one week of adaptive feeding, six-week-old male C57BL / 6J mice were fed a standard diet and drinking water. The Con group received a high-fat diet and high-sugar drinking water (23.1 g / L d-fructose + 18.9 g / L d-glucose) for 12 weeks to establish a NAFLD model. Twelve weeks later, mice in the Sily, Modified Fang-L, and Modified Fang-H groups were simultaneously administered the corresponding drug at a volume of 0.1 mL / 10 g. The drugs were dissolved in 0.5% CMC-Na. The remaining groups received the same dose of 0.5% CMC-Na for 12 weeks. After 12 weeks of drug administration, all groups were fasted for 12 hours. Blood was then collected by enucleation under isoflurane anesthesia. Following blood collection, the mouse hearts were perfused with 30 mL of normal saline, and the livers were subsequently removed.

[0038] Index detection:

[0039] 1. Oral glucose tolerance test

[0040] One week before sampling, six mice were randomly selected from each group and fasted for 12 hours before the experiment. During this fasting period, mice were deprived of food but not water. Based on their body weight, they were injected with a 20% glucose solution at a dose of 2.0 g / kg. After disinfecting the mouse tail with an alcohol swab, blood was drawn using a lancet. Blood glucose levels were measured and recorded at 0 min (fasting blood glucose), 15 min, 30 min, 60 min, 90 min, and 120 min. The area under the curve (AUC) was calculated using GraphPad Prism 9.0 software.

[0041] 2. Determination of body fat percentage and liver index of mice

[0042] Before sampling, based on the principle of nuclear magnetic resonance, a small nuclear magnetic analyzer was used to obtain the body fat-related values ​​of the mice.

[0043] After the autopsy, six mice were randomly selected from each group. Their body weight and liver wet weight were measured using an electronic balance. The organ index of each organ was calculated using the organ index formula: organ index = mouse organ fresh weight / mouse body weight × 100%.

[0044] 3. Four serum lipids, liver function indicators alanine aminotransferase, aspartate aminotransferase test

[0045] After drug intervention in NAFLD model mice, the levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol in the mice's serum were determined by testing four blood lipids, and the levels of serum alanine aminotransferase and aspartate aminotransferase were detected using kits.

[0046] 4. Serum fasting insulin and fibrosis index detection

[0047] After drug intervention, serum fasting insulin (FINS) levels in NAFLD model mice were measured by ELISA. The insulin resistance index was calculated using the homeostatic insulin resistance index (HOMA-IR): HOMA-IR = FINS × FBG / 22.5. Serum levels of fibrosis markers HA, Col-IV, and LN were also measured by ELISA.

[0048] 5. Detection of TG, MDA and SOD in liver tissue

[0049] After drug intervention in NAFLD model mice, the levels of total cholesterol, triglyceride, malondialdehyde and superoxide dismutase in the mouse liver tissue homogenate were determined according to the kit instructions.

[0050] 6. Histopathological examination

[0051] Liver tissue was collected and fixed in 4% paraformaldehyde for 24 hours. The tissue was then dehydrated using graded ethanol and xylene, embedded in paraffin, and sectioned at 4 μm thickness. Hematoxylin and eosin (HE) and picrosirius red staining were performed according to the manufacturer's instructions. Frozen liver sections were dehydrated in sucrose solution and then stained with Oil Red O by incubation for 30 minutes. The sections were then observed and photographed under a microscope.

[0052] 7. Liver histomorphological evaluation (NAS score)

[0053] The HE-stained images of liver tissue sections of mice in each group were scored according to the NAS scoring system developed by the Pathology Working Group of the American Association for Clinical Research on Nonalcoholic Fatty Liver Disease. Histological parameters included liver ballooning, steatosis, inflammation, and fibrosis, and the sum of the scores was summarized as the NAS score.

[0054] 8. Immunohistochemical staining

[0055] Liver tissue was fixed in 4% paraformaldehyde and embedded in paraffin blocks. 4-μm liver sections were prepared according to standard immunohistochemistry protocols. Liver sections were dehydrated and incubated with primary and secondary antibodies. The area of ​​positive staining was measured in a high-power field on each slide and quantified using Image J software.

[0056] 9. Quantitative Real-Time PCR

[0057] Total RNA was extracted from liver tissue and then reverse transcribed and quantitative real-time PCR (q-PCR) was performed using a real-time PCR system (LightCycler 480 Instrument II, Roche Diagnostics, Basel, BS, Switzerland) according to standard protocols. GAPDH was used as an internal control. Primer sequences are shown in Table 1.

[0058] Table 1. Primer sequences

[0059]

[0060] 10. Statistical analysis

[0061] GraphPad Prism 9.4.1 software was used for data analysis and graphing. Two-group comparisons were performed with a two-tailed unpaired t-test, and multiple group comparisons were performed with one-way ANOVA with Tukey's multiple comparison test. Data are expressed as mean ± standard error (x ± s). P <0.05 indicated that the difference was statistically significant.

[0062] Experimental results

[0063] 1. Body weight and food intake

[0064] The body weight of mice in each group Figure 1 , Table 3, food intake is shown in Table 2. Compared with the blank control group, the body weight of the model group increased significantly, and the food intake decreased significantly; compared with the model group, the body weight of the high-dose group of the modified formula decreased significantly without affecting the food intake.

[0065] Table 2. Food intake statistics of each group (n=6, )

[0066]

[0067] Note: Compared with the blank group: #### P <0.001.

[0068] Table 3. Weight statistics of each group (n=6, )

[0069]

[0070] Note: Compared with the blank group: ### P <0.001

[0071] 2. Liver appearance, liver index and body fat percentage

[0072] The appearance of the liver of mice in each group showed ( Figure 2 ), the livers of mice in the blank control group were dark brown and soft in texture; the liver volume of the model group was significantly increased, the liver surface was earthy white and the toughness was enhanced; the liver volume of the modified prescription group was significantly reduced compared with the model group, and the liver surface color of the modified prescription-H group turned light brown and the toughness was weakened; the liver volume of the Sily-positive control group was significantly reduced compared with the model group, and the liver surface color did not change much.

[0073] The results of liver index and body fat percentage measurement of mice in each group showed (Table 4) that compared with mice in the Con group, the liver index of mice in the Mod group was significantly increased, the fat content was significantly increased, and the muscle content was significantly decreased; compared with the Mod group, the liver index of the modified prescription and Sily groups were significantly decreased; the fat percentage of mice in the modified prescription group was significantly decreased, and the muscle percentage was significantly increased.

[0074] Table 4. Liver index and body fat percentage statistics of each group (n=6, )

[0075]

[0076] Note: Compared with the blank group: ### P <0.001; compared with the model group: * P <0.05, ** P <0.01.

[0077] 3. Oral glucose tolerance and insulin resistance

[0078] The results of OGTT and insulin resistance of mice in each group showed that ( Figure 3 (Table 5). Compared with the Con group, the Mod group showed significantly decreased oral glucose tolerance and increased insulin resistance. All other treatment groups showed significantly improved oral glucose tolerance and decreased insulin resistance compared with the Mod group. This suggests that the modified formula or Sily treatment can effectively improve glucose tolerance and insulin resistance in NAFLD mice.

[0079] Table 5. Statistics of glucose area under the curve, serum insulin content, and insulin resistance in each group (n=6, )

[0080]

[0081] Note: Compared with the blank group: ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

[0082] 4. Serum lipid levels and liver function damage

[0083] Results of the four blood lipid profiles (Table 6) showed that compared with the Con group, the Mod group had significantly increased serum TC, TC, and LDL-C levels, and significantly decreased HDL-C levels. After intervention with Sily and different doses of the modified formula, the mice had significantly decreased serum TC, TC, and LDL-C levels, while their HDL-C levels were significantly increased. This suggests that the modified formula can effectively improve the blood lipid profile of NAFLD mice, reducing serum TC, TG, and LDL-C levels and increasing HDL-C levels.

[0084] Table 6. Statistics of four blood lipids in each group (n=6, )

[0085]

[0086] Note: Compared with the blank group: ## P <0.01, ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

[0087] The results of liver function index measurement showed (Table 7) that the serum AST and ALT levels of mice in the Mod group were significantly higher than those in the Con group, indicating that the liver function of mice in the Mod group was damaged. Sily and different doses of the modified prescription administration groups could significantly reduce the serum AST and ALT levels of NAFLD mice induced by high fat and high sugar, and improve liver damage, among which the modified prescription group was better.

[0088] Table 7. Liver function index statistics of each group (n=6, )

[0089]

[0090] Note: Compared with the blank group: ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

[0091] 5. Pathological staining and NAS scoring

[0092] The results of H&E staining of liver tissue sections of mice in each group showed that ( Figure 4 ), the liver cells of the mice in the Mod group were swollen and rounded compared with those in the normal control group, with lipid droplet infiltration, ballooning and more intralobular inflammation; the liver plates of the mice in the Sily and different doses of the modified formula groups were arranged relatively neatly, the liver lobule structure was basically normal, with a small amount of lipid droplet infiltration and ballooning, the liver lobule inflammation was improved, and there was no obvious inflammatory cell accumulation foci.

[0093] H&E-stained liver sections from each group of mice were observed microscopically, and five randomly selected fields of view were evaluated for NAS pathology. The results, as shown in Table 8, showed significant increases in the liver steatosis index, ballooning index, and inflammatory cell infiltration index in the Mod mice, demonstrating typical NAFLD characteristics. NAS pathology scores in all treatment groups were significantly lower than those in the Mod group, demonstrating significant improvement. This suggests that Sily and the modified formula can improve NAFLD pathological features such as hepatic lipid accumulation, ballooning, and inflammatory cell infiltration.

[0094] Table 8. NAS scores of each group (n=6, )

[0095]

[0096] Note: Compared with the blank group: # P <0.05, ## P <0.01, ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

[0097] 6. Improvement of liver lipids

[0098] like Figure 5As shown, the Mod group mice showed extensive liver steatosis and lipid deposition (Table 9), with significantly elevated liver TG and TC levels (Table 10), indicating severe liver lipid accumulation. Administration of Sily and different doses of the modified formula significantly improved liver lipid deposition and decreased liver TG and TC levels, indicating that the modified formula is effective in ameliorating hepatic steatosis in high-glucose and high-fat-induced NAFLD mice, with superior efficacy compared to the silymarin group.

[0099] Table 9. Statistics of lipid droplet area in each group (n=3, x±s)

[0100]

[0101] Note: Compared with the blank group: ### P <0.001; compared with the model group: ** P <0.01.

[0102] Table 10. Statistical table of TG content in liver tissue of each group (n=3, )

[0103]

[0104] Note: Compared with the blank group: # P <0.05, ## P <0.01, ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

[0105] The results of gene expression related to lipid synthesis (Fasn, Srebp-1c, Scd1, Ppar-γ, ACC), transport (Cd36) and fatty acid oxidation (Cpt1a, Ppar-α, Atgl) showed (Table 11) that the modified formula and Sily treatment could significantly reduce the mRNA levels of Fasn, Srebp-1c, Scd1, Cd36, ACC and Ppar-γ in NAFLD mice, and reverse the abnormal downregulation of mRNA levels of Atgl, Cpt1α and Ppar-α, indicating that the modified formula can effectively improve the liver lipid metabolism disorder induced by high glucose and high fat in NAFLD mice.

[0106] Table 11. Statistics of mRNA levels of genes related to liver lipid metabolism in each group (n=6, )

[0107]

[0108] Note: Compared with the blank group: ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

[0109] 7. Improvement of inflammation

[0110] F4 / 80 immunohistochemistry results showed that ( Figure 6 , Table 12). Compared with the Con group, the number of F4 / 80-positive cells in the livers of mice in the Mod group increased significantly, and the level of hepatic macrophage infiltration was significantly enhanced. Compared with the Mod group, the number of F4 / 80-positive cells in the livers of mice in the modified formula group decreased significantly, indicating that the modified formula can significantly inhibit macrophage infiltration in the livers of mice. Real-time quantitative PCR was used to detect the gene expression of inflammatory factors in the livers of mice in each group. The results are shown in Table 13. Compared with the Mod group, long-term oral administration of the modified formula significantly inhibited the gene expression of TNF-α, IL-1β, and MCP-1 in NAFLD mice, showed a trend of suppressing the gene expression of the pro-inflammatory factor IL-6, and significantly activated the gene expression of the anti-inflammatory factor IL-10 (Table 13), indicating that the modified formula has an ameliorative effect on the hepatic inflammatory response in NAFLD mice.

[0111] Table 12. Statistics of F4 / 80 immunohistochemical staining positive areas in liver sections of each group (n=3, )

[0112]

[0113] Note: Compared with the blank group: ### P <0.001; compared with the model group: ** P <0.01.

[0114] Table 13. Statistics of mRNA levels of genes related to liver inflammatory factors in each group (n=6, )

[0115]

[0116] Note: Compared with the blank group: ## P <0.01, ### P <0.001; compared with the model group: *P <0.05, ** P <0.01, *** P <0.001.

[0117] 8. Improvement of fibrosis

[0118] like Figure 7 As shown in Table 14, the livers of mice in the Mod group showed significant fibrotic lesions compared to those in the Con group, with a significant increase in collagen fibers, concentrated around the portal areas and around degenerated hepatocytes. Treatment with Sily and different doses of the modified formula significantly reduced collagen fiber deposition in NAFLD mice. Furthermore, the modified formula significantly reduced serum levels of fibrosis markers LN and HA (Table 15). Compared with the Con group, the mRNA expression of pro-fibrotic markers was significantly upregulated in the Mod group. Administration of the modified formula significantly inhibited the gene expression of α-SMA, Col1a1, FN-1, and Tgf-β1 in the livers of NAFLD mice (Table 16). This indicates that the modified formula significantly improves liver fibrosis in induced NAFLD mice.

[0119] Table 14. Statistics of collagen fibers in liver sections of each group (n=3, )

[0120]

[0121] Note: Compared with the blank group: ### P <0.001; compared with the model group: *** P <0.001.

[0122] Table 15. Statistics of serum fibrosis factor levels in each group (n=6, )

[0123]

[0124] Note: Compared with the blank group: # P<0.05, ## P <0.01, ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

[0125] Table 16. Statistics of mRNA levels of liver fibrosis-related genes in each group (n=6, )

[0126]

[0127] Note: Compared with the blank group: ### P <0.001; compared with the model group: ** P <0.01, *** P <0.001.

[0128] 9. Improvement of oxidative stress

[0129] The results of the oxidative stress levels in the liver tissues of each group showed (Table 17) that compared with the Con group, the MDA level in the liver of the mice in the Mod group was significantly increased, the SOD level was significantly decreased, and the oxidative stress in the liver of the mice in the Mod group was enhanced; treatment with Sily and different doses of the modified prescription significantly reduced the MDA level in the liver of the NAFLD mice induced by high sugar and high fat, increased the SOD level, and improved oxidative stress.

[0130] Table 17. Statistics of oxidative stress levels in liver tissues of each group (n=6, )

[0131]

[0132] Note: Compared with the blank group: # P <0.05, ## P <0.01, ### P <0.001; compared with the model group: * P <0.05, ** P <0.01, *** P <0.001.

Claims

1. A Chinese medicine composition for preventing or treating non-alcoholic fatty liver disease, characterized in that: The invention is composed of the following ingredients: 2-30 parts of ginseng, 1-30 parts of malt, 3-40 parts of poria cocos, 3-50 parts of buzia leaves, 3-30 parts of selfheal and 2-30 parts of sophora japonica flowers.

2. The Chinese medicine composition according to claim 1, characterized in that: The weight proportions of the ingredients are as follows: 3 parts of ginseng, 6 parts of malt, 6 parts of Poria cocos, 7.5 parts of Buza leaves, 4.5 parts of Prunella vulgaris and 3 parts of Sophora japonica flowers.

3. The method for preparing the Chinese medicine composition according to claim 1, characterized in that: The method comprises the following steps: preparing the following raw materials in parts by weight: 2-30 parts of ginseng, 1-30 parts of malt, 3-40 parts of poria cocos, 3-50 parts of buzia leaves, 3-30 parts of selfheal and 2-30 parts of sophora japonica flowers; adding water, heating and refluxing for extraction twice, filtering, combining the two extracts, concentrating and drying.

4. The method for preparing the Chinese medicine composition according to claim 3, wherein: During the extraction process, the mass-to-volume ratio of the material to the liquid (g / mL) was 1:3 to 1:20; each extraction time was 1 to 3 hours.

5. The method for preparing the Chinese medicine composition according to claim 4, wherein: During the first extraction, the mass-to-volume ratio of the material to the liquid (g / mL) was 1:10; during the second extraction, the mass-to-volume ratio of the material to the liquid (g / mL) was 1:8; each extraction time was 1 h.

6. Use of the traditional Chinese medicine composition according to claim 1 or 2 or the traditional Chinese medicine composition prepared by the method according to any one of claims 3 to 5 in the preparation of a medicament for treating or preventing non-alcoholic fatty liver disease.

7. A drug for preventing or treating non-alcoholic fatty liver disease, characterized in that: The invention relates to a Chinese medicine composition according to claim 1 or 2 or a Chinese medicine composition prepared by the method according to any one of claims 3 to 5.

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