Use of gouteng in preparation of a medicine for preventing and / or treating metabolic-associated steatohepatitis (MASH)

Abstract

The application discloses application of Gomphnae in preparation of a medicine for preventing and / or treating metabolic associated steatohepatitis (MASH). The application proves by animal experiments that Gomphnae can significantly improve abnormal body weight, abnormal liver weight, liver steatosis, ballooning, inflammatory infiltration and liver fibrosis in a MASH model of a mouse induced by AMLN diet with high fat, high fructose and high cholesterol, reduce serum ALT and AST levels, and down-regulate expression of inflammation-related genes such as TNF-alpha, IL-1 beta , Collagen I , Collagen III , CTGF fibrosis-related genes in liver tissue, and reduce expression of inflammation or fibrosis-related proteins such as F4 / 80, IL-1β, α-SMA and Collagen I, which can effectively relieve and / or treat MASH through mechanisms such as regulation of lipid metabolism, inhibition of inflammation signal pathways and fibrosis signal pathways. The application provides a medicine selection from a natural plant for prevention and treatment of MASH, and has a good development prospect.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to the application of a natural medicinal plant in pharmaceutical manufacturing, and particularly to the application of *Gynostemma pentaphyllum* in the preparation of a drug for the prevention and / or treatment of metabolic dysfunction-associated steatohepatitis (MASH). Background Technology

[0002] Metabolic dysfunction-associated fatty liver disease (MASLD) is a group of chronic liver diseases characterized by abnormal lipid deposition in the liver, accompanied by cardiometabolic risk factors such as obesity, abnormal glucose metabolism, hypertension, or dyslipidemia. It has become one of the most common chronic liver diseases worldwide. Its disease spectrum includes simple hepatic steatosis and its progressive form—metabolic dysfunction-associated steatohepatitis (MASH). MASH is characterized by hepatocellular steatosis, ballooning degeneration, lobular inflammation, and varying degrees of fibrosis, and can further develop into cirrhosis, hepatocellular carcinoma, and even liver failure.

[0003] In recent years, Resmetirom and Wegovy (semaglutide) have been approved by the FDA for adult patients with moderate to advanced liver fibrosis (MASH), indicating that MASH treatment has entered the drug intervention stage. However, existing drugs still have problems such as limited applicable population, need for long-term use, gastrointestinal adverse reactions, cost, and insufficient accessibility. The occurrence and development of MASH involves multiple stages, including hepatic lipid accumulation, hepatocellular damage, inflammatory response, fibrosis progression, and systemic metabolic disorders. Current treatment methods are still insufficient to fully meet the needs of comprehensive intervention for the multi-stage pathological changes in MASH. Therefore, it is necessary to develop a new drug or drug use that can improve the pathological changes related to hepatic steatosis, inflammatory response, and fibrosis in MASH patients.

[0004] Traditional Chinese medicine and its extracts are characterized by multi-component and multi-stage intervention, and have potential application value in lipid metabolism regulation, anti-inflammation, and anti-fibrosis. (Ganmutong (Ganmutong)...) Clematis loureiroana DC. is a genus of Clematis in the family Ranunculaceae. Clematis Clematis chinensis, also known as silk clematis, is a plant listed in the *Chinese Materia Medica* and is a traditional medicinal herb native to Guangdong and Guangxi provinces. It has a sweet taste and slightly cooling properties, possessing sedative, analgesic, and blood pressure-lowering effects. Clinically, it is mainly used to lower blood pressure and treat various complications caused by hypertension, with relatively few side effects. Current research primarily focuses on the effects of Clematis chinensis or its extracts on lowering blood pressure, myocardial protection, antioxidation, and anti-apoptosis, but has not yet linked it to specific pathological processes in MASH such as fatty degeneration, inflammatory response, and fibrosis.

[0005] To date, no published literature or patents have demonstrated that *Gynostemma pentaphyllum* is used to prevent and treat metabolic dysfunction-associated steatohepatitis (MASH). Existing technologies mainly focus on the effects of total flavonoids from *Gynostemma pentaphyllum* on protecting damaged cells, resisting myocardial ischemia, and preventing arrhythmia, but they have not clearly linked them to the specific and complex pathological process of MASH (steatodegeneration + inflammation + ballooning degeneration), and there is a lack of in-depth efficacy verification and mechanism exploration in animal models of MASH. Summary of the Invention

[0006] To address the shortcomings of existing MASH treatments mentioned above in terms of applicability, safety, long-term medication adherence, and multi-stage pathological intervention, this invention provides the following technical solution: The use of Gynostemma pentaphyllum in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-associated steatohepatitis (MASH).

[0007] Furthermore, the drug has Aristolochia debilis as its main active ingredient.

[0008] Furthermore, the drug uses Gynostemma pentaphyllum as its sole active ingredient.

[0009] Furthermore, the drug also includes pharmaceutically commonly used excipients or auxiliary ingredients.

[0010] Furthermore, the *Ganmutong* mentioned is the original medicinal material of *Ganmutong*.

[0011] Furthermore, the *Gynostemma pentaphyllum* is *Gynostemma pentaphyllum* powder.

[0012] Furthermore, the dosage form of the drug is an oral formulation.

[0013] Furthermore, the oral preparation includes any one of tablets, capsules, granules, powders, oral liquids, or pills.

[0014] Furthermore, the metabolic dysfunction-related steatohepatitis is metabolic dysfunction-related steatohepatitis accompanied by liver fibrosis.

[0015] The beneficial effects of this invention are as follows: 1. This invention is the first to discover that Gynostemma pentaphyllum can be used to prepare drugs for the prevention and / or treatment of metabolic dysfunction-associated steatohepatitis (MASH), thus expanding the pharmaceutical use of Gynostemma pentaphyllum in this indication.

[0016] 2. After intervention with Gynostemma pentaphyllum, the body weight and liver weight of MASH model mice induced by AMLN diet were significantly reduced, the liver tissue structure was significantly improved, and liver lipid deposition, inflammatory cell infiltration, ballooning degeneration and collagen deposition were effectively improved.

[0017] 3. Ganmutong can protect the liver by improving abnormal metabolic indicators such as TC and TG in plasma and liver tissue of MASH model mice and reducing plasma ALT and AST levels.

[0018] 4. Ganmutong can lower... Srebp1, FASN, SCD1 Increasing the expression of lipid synthesis-related genes improves hepatic steatosis; while simultaneously downregulating... TNF-α, MCP-1, IL-1β Inflammation-related genes and Col1α1, Col3α1, CTGF The expression of fibrosis-related genes can reduce liver inflammation and fibrotic changes.

[0019] 5. Ganmutong can reduce the number of F4 / 80 positive cells in the liver tissue of MASH model mice and reduce the intensity of IL-1β positive staining, indicating that Ganmutong can reduce the infiltration of hepatic macrophages and inhibit the protein expression of the pro-inflammatory factor IL-1β, thereby alleviating the MASH-related hepatic immune inflammatory response.

[0020] 6. Ganmutong can reduce the expression levels of α-SMA and COL1α1 fibrosis-related proteins in the liver tissue of MASH model mice. In particular, high-dose Ganmutong can significantly inhibit the abnormal upregulation of α-SMA and COL1α1 proteins, indicating that Ganmutong can inhibit the abnormal expression of some liver fibrosis-related proteins at the protein level, thereby improving MASH-related liver fibrosis.

[0021] 7. Ganmutong can inhibit the NF-κB inflammatory signaling pathway and TGF-β / Smad fibrosis-related molecular changes, thereby improving hepatic steatosis, inflammatory response and liver fibrosis in MASH model mice, achieving the effect of preventing and / or treating metabolic dysfunction-related steatohepatitis. Attached Figure Description

[0022] Figure 1 These are the mass spectrometry analysis results of some chemical components in Aristolochia debilis in this application; Figure 2 The following are images illustrating the therapeutic effects of *Gynostemma pentaphyllum* on MASH model mice in this application: (A: Representative morphological photographs and liver images of mice in each group at the experimental endpoint; B: Quantitative analysis results of body weight and liver weight of mice in each group; C: Representative images of H&E staining, Oil Red O staining, Masson staining, and Sirius Red staining of liver tissue from mice in each group (magnification: 100X); n=10 / group;* P <0.05,** P <0.01, *** P <0.001, **** P <0.0001); Figure 3The effects of *Gynostemma pentaphyllum* on metabolism and liver function in MASH model mice as described in this application are as follows: (A: Plasma levels of total cholesterol (TC) and triglycerides (TG) in each group of mice; B: Liver tissue levels of total cholesterol (TC) and triglycerides (TG) in each group of mice; C: Plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in each group of mice; n=10 / group;* P <0.05,** P <0.01, **** P <0.0001); Figure 4 The effects of *Gynostemma pentaphyllum* on the expression of liver-related genes in MASH model mice in this application are as follows: (A: Expression levels of lipid synthesis-related genes in the liver of each group of mice; B: Expression levels of inflammation-related genes in the liver of each group of mice; C: Expression levels of fibrosis-related genes in the liver of each group of mice; n=10 / group;*) P <0.05,** P <0.01, *** P <0.001, **** P <0.0001); Figure 5 Immunohistochemical staining images of the effect of Aristolochia debilis on the expression of hepatic inflammatory markers in MASH model mice in this application (A: representative images of F4 / 80 immunohistochemical staining in liver tissue of mice in each group; B: representative images of IL-1β immunohistochemical staining in liver tissue of mice in each group; n=10 / group); Figure 6 This application describes the effects of *Gynostemma pentaphyllum* on the expression of liver fibrosis-related proteins in MASH model mice (A: Western blotting results of α-SMA and COL1α1 proteins in the liver of each group of mice; B: Expression levels of α-SMA protein relative to the internal control β-actin protein in the liver of each group of mice; C: Expression levels of COL1α1 protein relative to the internal control β-actin protein in the liver of each group of mice; **** P <0.0001); Figure 7 This application describes the effects of *Gynostemma pentaphyllum* on the expression of proteins in liver-related signaling pathways in MASH model mice. (A: Western blotting results of proteins in the NF-κB and TGF-β / Smad pathways, which are inflammation-related signaling pathways, in the livers of mice in each group; B: The ratio of p-p65 to p65 expression levels in the livers of mice in each group; C: The ratio of p-IκBα to IκBα expression levels in the livers of mice in each group; D: The ratio of p-Smad2 / 3 to Smad2 / 3 expression levels in the livers of mice in each group; ****) P<0.0001). Detailed Implementation

[0023] To facilitate a clearer understanding of the objectives, technical solutions, and advantages of this invention, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of this application and are not intended to limit this application. On the contrary, the purpose of providing these embodiments is to enable a more thorough and comprehensive understanding of the disclosure of this invention. Unless otherwise specified, the reagents and methods used in this invention are conventional reagents and methods in this technical field. Unless otherwise specified, the reagents and materials used in the following embodiments are commercially available.

[0024] Materials Methods 1.1 Material Preparation Ganmutong ( Clematis loureiroana After collection (DC.), the leaves were dried, impurities were removed, washed, pulverized, and passed through a 200-mesh sieve to obtain *Gynostemma pentaphyllum* powder for later use. Chemical composition analysis of *Gynostemma pentaphyllum* was performed using ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), revealing monoterpenes, sesquiterpenes, aldehydes, ketones, alkenes, phenols, and alcohols. It was particularly rich in flavonoids such as norihesperidin, rutin, baicalin, euryptin, quercetin, and morin D. Figure 1 (As shown).

[0025] Resmetirom (lot number: HY-2216) was purchased from MedChemExpress.

[0026] MASH modeling feed (batch number: AMLN, high trans fat (40 kcal%), high cholesterol (2%), high fructose (22%) rat food) and growth feed (item number: LF10C, 10 kcal% fat and low fat rat food) were purchased from Detz Biotechnology (Wuxi) Co., Ltd.

[0027] Hematoxylin (batch number: G1080), eosin (batch number: G1100), modified Masson trichrome staining kit (batch number: G1346), and modified Sirius red staining kit (batch number: G1472) were purchased from Solarbio.

[0028] Saturated Oil Red O staining solution (batch number: G1260) was purchased from Servicebio.

[0029] Alanine aminotransferase (ALT, batch number: C009 2 1), aspartate aminotransferase (AST, batch number: C0102 1), triglycerides (TG, batch number: A110 1 1), and total cholesterol (TC, batch number: A111 1 1) were purchased from Nanjing Jiancheng Biotechnology Institute.

[0030] The inventors designed using NCBI and Primer Blast respectively. Srebp1, FASN, SCD1, TNF-α, MCP-1, IL-1 β, Col3α1, Col1α1, CTGF and RNA18S1 The primers are shown in Table 1 below. The primers in Table 1 were synthesized by Sangon Biotech (Shanghai) Co., Ltd.

[0031] Table 1 Primer sequences for the target gene

[0032] Antibodies against nuclear factor κB inhibitor α (IκBα, batch number: 4814), phosphorylated nuclear factor κB inhibitor α (p-IκBα, batch number: 9246), p65 (batch number: 3033), phosphorylated p65 (p-p65, batch number: 4764), Smad homolog 2 / 3 (Smad2 / 3, batch number: 8685S), phosphorylated Smad homolog 2 / 3 (p-Smad2 / 3, batch number: 8828S), interleukin-1β (IL-1β, batch number: 12703T), HRP-labeled goat anti-rabbit IgG (batch number: 7074P2), and HRP-labeled horse anti-mouse IgG (batch number: 7076S) were purchased from Cell Signaling. Technology; the antibody for β-actin (batch number: AC026) was purchased from ABclonal Company in Wuhan, China; the antibody for mouse growth factor-like motif mucin-like hormone-like receptor (F4 / 80, batch number: sc-377009) was purchased from Santa Cruz Company.

[0033] 1.2 Animal Research Fifty male C57BL / 6J mice aged 6-8 weeks were purchased from Zhuhai Beston Biotechnology Co., Ltd. and housed in an SPF-grade animal experimental center under the following conditions: room temperature 22-25℃, 12-hour day-night cycle, 55% relative humidity, and free access to water and food.

[0034] After one week of acclimatization, 50 mice were randomly divided into five groups: a normal control group, a MASH model group, a low-dose Ganmutong group, a high-dose Ganmutong group, and a positive control group (Resmetirom), with 10 mice in each group. Except for the normal control group, the other groups of mice were fed an internationally recognized high-fat, high-sugar, and high-cholesterol AMLN diet (40% fat, 2% cholesterol, and 22% fructose) for 20 weeks to induce the MASH model.

[0035] Ganmutong was administered concurrently from the start of modeling. The low- and high-dose Ganmutong groups were given 70 mg / kg / day and 140 mg / kg / day by gavage, respectively (Ganmutong powder dissolved in 0.5% carboxymethyl cellulose sodium solution (0.5% CMC-Na)). The positive control group was given Resmetirom 3 mg / kg / day by gavage (dissolved in 0.5% CMC-Na). The normal control group and the model group were given an equal volume of 0.5% CMC-Na. The administration was continued for 20 weeks.

[0036] 1.3 Detection Method (1) Biochemical index detection methods Blood plasma or liver tissue biochemical parameters, including ALT, AST, TG, and TC, should be tested using commercially available kits according to the manufacturer's instructions.

[0037] (2) Histological analysis and Oil Red O staining detection method Fresh mouse liver tissue was collected, fixed in 4% neutral paraformaldehyde solution, dehydrated, cleared, and embedded in paraffin to prepare paraffin sections with a thickness of approximately 5 μm. The sections were stained according to the instructions of hematoxylin-eosin (H&E) staining solution, modified Masson's trichrome staining kit, and modified Sirius red staining kit. Separately, fresh mouse liver tissue was embedded in OCT and frozen to prepare sections with a thickness of approximately 7 μm. These sections were stained according to the instructions of Oil Red O staining kit. The sections were mounted with glycerol to obtain information on lipid droplet accumulation in the liver tissue sections and cells. The sections were observed and photographed under an optical microscope.

[0038] (3) Quantitative real-time PCR detection method 40 mg of mouse liver tissue was ground and total RNA was extracted using an RNA extraction kit (LS1040, Promega). Reverse transcription was then performed using a cDNA reverse transcription kit (11141ES60, Yeasen) according to the manufacturer's instructions. The reverse transcription conditions were: 25℃ for 5 min, 55℃ for 15 min, and 85℃ for 5 min. qRT-PCR was then performed using a qRT-PCR kit (11201ES08, Yeasen). The qRT-PCR program was: 95℃ for 5 min, 95℃ for 10 s, 60℃ for 20 s, and 72℃ for 35 s, for 40 cycles, with a 2... - Ct Relative quantification method for mRNA calculation. Target gene ( Srebp1, FASN, SCD1, TNF-α, MCP-1, IL-1β, Col3α1, Col1α1, CTGF ) and internal reference genes RNA18S1 The primer sequences are shown in Table 1 and were synthesized by Sangon Biotech (Shanghai) Co., Ltd.

[0039] (4) Immunoblotting detection method Total protein was extracted from mouse liver tissue using a lysis buffer containing RIPA, protease inhibitors, and phosphatase inhibitors; protein concentration in mouse liver tissue was determined using a BCA protein assay kit (A55865, Thermo Scientific).

[0040] The mixture of equal volumes of protein sample, RIPA solution, and loading dye in a specific ratio was loaded onto 10% SDS. The target protein was separated on a PAGE gel and migrated to a polyvinylidene fluoride (PVDF) membrane according to a standard procedure. After blocking with 5% BSA for 1 hour, the target protein was incubated overnight at 4°C with primary antibodies. The primary antibodies included COL1α1 (1:1000), α-SMA (1:1000), p65 (1:1000), p-p65 (1:1000), IκBα (1:1000), p-IκBα (1:1000), TGF-β1 (1:1000), Smad2 / 3 (1:1000), and p-Smad2 / 3 (1:1000), with β-actin (1:1000) used as an internal control protein. In addition, the samples were incubated at room temperature for 1 hour with HRP-labeled goat anti-rabbit IgG (1:5000) or horse anti-mouse IgG (1:5000), respectively. The protein bands were then captured by adding an ECL Advanced kit (GE Biosciences) and analyzed for grayscale values ​​using ImageJ software.

[0041] (5) Statistical analysis Statistical analysis was performed using GraphPad Prism 8.0 software. All experiments were repeated at least three times, and results are expressed as mean ± standard deviation (X ± SD). P Values ​​<0.05 are considered statistically significant.

[0042] Example 1: The effect of Aristolochia debilis on improving general physical signs and liver pathological morphology in diet-induced MASH model mice with AMLN. This study induced the MASH model using an AMLN diet (40% fat, 2% cholesterol, 22% fructose). After 20 weeks of administration, mice were euthanized according to animal welfare guidelines, and liver tissue was collected. The liver tissue was stained with H&E, Masson's stain, Sirius Red stain, and Oil Red O stain.

[0043] like Figure 2As shown in Figure A, compared with the normal control group mice, the model group mice were significantly obese, with pale yellow livers and significantly increased liver volume, indicating that the mice exhibited significant lipid metabolism abnormalities and liver lipid accumulation after AMLN dietary induction, proving the successful establishment of the MASH model. Compared with the model group, the low-dose Ganmutong group, the high-dose Ganmutong group, and the positive control group mice showed improved body size, liver color returned to a uniform reddish-brown, and liver volume decreased, suggesting that Ganmutong can improve the gross morphology of the liver and lipid accumulation in MASH mice.

[0044] like Figure 2 As shown in B, analysis of mouse body weight revealed that the high-dose Ganmutong group had significantly lower body weight than the model group ( P <0.001, the positive control group was significantly lower than the model group ( P <0.0001), and there was no statistically significant difference between the two groups ( P >0.05), indicating that Gynostemma pentaphyllum can improve the abnormal body weight in MASH model mice, and the effect of high-dose Gynostemma pentaphyllum on improving the body weight of MASH model mice is comparable to that of the positive control. Analysis of mouse liver weight showed that the liver weight of mice in both the high-dose Gynostemma pentaphyllum group and the positive control group was significantly lower than that in the model group ( P <0.0001). In summary, the high-dose group of Ganmutong can significantly improve the abnormal body weight and liver weight in MASH model mice, and its effect on improving body weight is comparable to that of the positive control drug (Resmetirom).

[0045] like Figure 2 As shown in Figure C, H&E staining of liver tissue revealed significant fatty degeneration, inflammatory cell infiltration, and ballooning degeneration in the liver tissue of the model group mice. Compared with the model group, the pathological changes in the liver tissue of mice in the low-dose Ganmutong group, high-dose Ganmutong group, and positive control group were significantly reduced, and the severity of the aforementioned lesions was significantly improved. Oil Red O staining showed a significant increase in the area of ​​lipid deposition in the liver of the model group; lipid deposition was significantly reduced in the low-dose Ganmutong group, high-dose Ganmutong group, and positive control group, suggesting that Ganmutong can alleviate lipid accumulation in the liver of MASH mice induced by AMLN diet. Masson staining and Sirius Red staining showed obvious collagen deposition and fibrosis in the liver tissue of the model group; the area of ​​collagen deposition was significantly reduced in the liver tissue of mice in the low-dose Ganmutong group, high-dose Ganmutong group, and positive control group, and the degree of fibrosis was significantly improved.

[0046] Figure 2 The experimental data corresponding to B are shown in Table 2.

[0047] Table 2 Summary of mouse body weight and liver weight data

[0048] As shown in Table 2, in terms of mouse body weight, the model group increased by 16.7% compared with the normal control group, the low-dose Ganmutong group decreased by 6.7% compared with the model group, and the high-dose Ganmutong group decreased by 13.1% compared with the model group; in terms of mouse liver weight, the model group increased by 50.0% compared with the normal control group, the low-dose Ganmutong group decreased by 16.0% compared with the model group, and the high-dose Ganmutong group decreased by 28.2% compared with the model group.

[0049] Therefore, it is evident that intervention with Gynostemma pentaphyllum significantly improved abnormal body weight and liver weight in MASH model mice, reduced hepatic lipid accumulation, improved liver tissue structure, and decreased the degree of steatosis, inflammatory infiltration, ballooning degeneration, and fibrosis. These results demonstrate that Gynostemma pentaphyllum can alleviate and / or treat MASH-related liver pathological damage without causing discomfort, death, deformity, or carcinogenesis in the model mice, thus confirming the safety of Gynostemma pentaphyllum and providing potential for its use in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-related steatohepatitis.

[0050] Example 2: The regulatory effect of Aristolochia debilis on blood lipids, liver lipids and transaminase levels in MASH model mice This embodiment further examines the regulatory effects of Gynostemma pentaphyllum on the levels of blood lipids (plasma TC, TG), liver lipids (liver TC, TG), and liver function transaminases (plasma ALT, AST) in MASH model mice.

[0051] This study induced a MASH model using an AMLN diet (40% fat, 2% cholesterol, 22% fructose). After 20 weeks of administration, mice were euthanized according to animal welfare guidelines, and blood and liver tissue were collected. Plasma was separated from the mouse blood to detect the levels of total cholesterol (TC), triglycerides (TG), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). Liver tissue was collected and homogenized to detect the levels of total cholesterol (TC) and triglycerides (TG) in the liver tissue.

[0052] like Figure 3 As shown in Figure A, compared with the normal control group, the plasma total cholesterol (TC) level in the model group mice was significantly increased ( P <0.0001), while plasma triglyceride (TG) levels were significantly reduced ( PThe change was <0.0001), which is consistent with the characteristic lipid profile of the AMLN diet-induced MASH model. Compared with the plasma TC level in the model group mice, the plasma TC levels in the low-dose Ganmutong group, high-dose Ganmutong group, and positive control group were significantly reduced, suggesting that Ganmutong can effectively reduce the abnormally elevated plasma TC level in MASH model mice and improve dyslipidemia. Compared with the plasma TG level in the model group mice, the plasma TG levels in the low-dose Ganmutong group, high-dose Ganmutong group, and positive control group were increased to varying degrees, suggesting that Ganmutong intervention has a certain regulatory effect on the plasma TG level in AMLN diet-induced MASH model mice.

[0053] like Figure 3 As shown in B, the levels of TC and TG in the liver of mice in the model group were significantly higher than those in the normal control group. P <0.0001 indicates that a large amount of lipid accumulation occurred in the liver of mice after AMLN dietary induction, further proving the successful establishment of the MASH model. Compared with the model group, the levels of TC and TG in the liver of mice in the low-dose group, high-dose group, and positive control group of Ganmutong were significantly reduced, and both dose groups of Ganmutong showed obvious dose dependence. Among them, there was no statistically significant difference between the high-dose group and the positive control group in reducing liver TG content ( P The value >0.05 indicates that both drugs are equally effective in improving this indicator. These results demonstrate that *Gynostemma pentaphyllum* can effectively reduce lipid accumulation in the liver tissue of MASH model mice and improve MASH-related hepatic steatosis.

[0054] like Figure 3 As shown in C, compared with the normal control group, the plasma ALT and AST levels in the model group mice were significantly increased ( P The result (<0.0001) indicates that mice induced by the AMLN diet showed significant liver function damage. Compared with the model group, the plasma ALT and AST levels of mice in the low-dose group, high-dose group, and positive control group were significantly reduced, and the reducing effect of Glycyrrhiza uralensis on ALT and AST was dose-dependent. Among them, the high-dose group of Glycyrrhiza uralensis showed the most significant effect in reducing ALT and AST. These results indicate that Glycyrrhiza uralensis can effectively improve hepatocellular damage and abnormal liver function in MASH model mice.

[0055] Figure 3 A to Figure 3 The experimental data corresponding to C are shown in Table 3.

[0056] Table 3 Summary of mouse plasma TC, TG, ALT and AST, and liver TC and TG data

[0057] Table 3 shows that, regarding the plasma TC level in mice, the model group was 69.8% higher than the normal control group, the low-dose Ganmutong group was 18.1% lower than the model group, and the high-dose Ganmutong group was 20.2% lower than the model group. Regarding the plasma TG level in mice, the model group was 64.0% lower than the normal control group, the low-dose Ganmutong group was 144.4% higher than the model group, the high-dose Ganmutong group was 61.1% higher than the model group, and the positive control group was 66.7% higher than the model group. Regarding the plasma ALT level in mice, the model group was 501.7% higher than the control group, the low-dose Ganmutong group was 63.1% lower than the model group, the high-dose Ganmutong group was 76.0% lower than the model group, and the positive control group was 66.5% lower than the model group. Regarding the plasma AST level in mice, the model group was 139.0% higher than the normal control group, the low-dose Ganmutong group was 15.6% lower than the model group, the high-dose Ganmutong group was 45.1% lower than the model group, and the positive control group was 37.5% lower than the model group.

[0058] Regarding the total cholesterol (TC) level in the liver of mice, the model group was 141.2% higher than the normal control group, the low-dose Ganmutong group was 9.8% lower than the model group, and the high-dose Ganmutong group was 24.4% lower than the model group. Regarding the triglyceride (TG) level in the liver of mice, the model group was 159.4% higher than the normal control group, the low-dose Ganmutong group was 51.8% lower than the model group, the high-dose Ganmutong group was 76.0% lower than the model group, and the positive control group was 72.8% lower than the model group.

[0059] Therefore, it is evident that Gynostemma pentaphyllum can improve dyslipidemia, hepatic lipid accumulation, and liver function damage in MASH model mice, indicating its efficacy in alleviating and / or treating MASH-related lipid metabolism disorders and liver function damage. Specifically, high-dose Gynostemma pentaphyllum is comparable to or better than the positive control drug (Resmetirom) in reducing hepatic triglycerides (TG) and improving plasma transaminase levels (ALT, AST).

[0060] Example 3: Regulatory effects of Aristolochia debilis on the expression of genes related to liver lipid metabolism, inflammation, and fibrosis in MASH model mice. This embodiment further employs quantitative real-time PCR (qRT-PCR) to detect the mRNA expression levels of genes related to lipid synthesis, inflammation, and fibrosis in mouse liver tissue. Among these, the lipid synthesis-related genes are... Srebp1, FASN, SCD1 Inflammation-related genes are TNF-α, MCP-1, IL-1β ; fibrosis-related genes are Col3α1, Col1α1, CTGF .

[0061] like Figure 4 As shown in Figure A, compared with the normal control group, the model group mice had lipid synthesis-related genes in their liver tissue. Srebp1, FASN and SCD1The relative expression levels of mRNA were all significantly upregulated. P <0.0001 indicates that after AMLN dietary induction, the transcriptional programs related to fatty acid synthesis and lipid production in the liver of model group mice were significantly activated. Compared with the model group, the low-dose Ganmutong group, the high-dose Ganmutong group, and the positive control group all significantly downregulated the relative mRNA expression levels of the above genes ( P The result <0.0001 indicates that Gynostemma pentaphyllum can inhibit the abnormal upregulation of lipid synthesis-related genes in the liver of MASH model mice. Specifically, the downregulation of these genes in the high-dose Gynostemma pentaphyllum group was similar to that in the positive control group, with no statistically significant difference between the two. P >0.05). The above results indicate that Gynostemma pentaphyllum can significantly regulate the expression of genes related to liver lipid synthesis in MASH model mice, suggesting that it may improve liver lipid metabolism disorders in MASH model mice by inhibiting liver lipid production pathways.

[0062] like Figure 4 As shown in B, compared with the normal control group, the model group mice had higher levels of inflammation-related genes in their liver tissue. TNF-α, MCP-1 and IL-1β The relative expression levels of mRNA were significantly increased (of which TNF-α, MCP-1 of P <0.0001; IL-1β of P The value <0.001 indicates that inflammation-related transcription levels were significantly activated in the liver tissue of MASH model mice. Compared with the model group, the low-dose Ganmutong group, the high-dose Ganmutong group, and the positive control group all showed varying degrees of reduction. TNF-α, MCP-1 and IL-1β The expression of the high-dose licorice group was observed. MCP-1 and IL-1β The downregulation of the gene was slightly better than that of the positive control group, indicating that Ganmutong can inhibit the abnormal upregulation of liver inflammation-related genes in MASH model mice, suggesting that it has the effect of improving MASH liver inflammation-related molecules.

[0063] like Figure 4 As shown in C, compared with the normal control group, the liver tissue of the model group mice had... Col3α1, Col1α1 and CTGF The relative expression levels of mRNA were all significantly increased ( P The value <0.0001 indicates that fibrosis-related molecular responses in the liver tissue of MASH model mice were significantly activated after AMLN dietary induction. Compared with the model group, the low-dose group, high-dose group, and positive control group of Gynostemma pentaphyllum all showed significant downregulation. Col3α1, Col1α1 and CTGF relative mRNA expression levels (targeting) Col3α1Genes, low-dose Ganmutong group relative to model group P <0.001, other genes P <0.0001. Among them, the downregulation of the above-mentioned fibrosis-related genes in the high-dose Ganmutong group was not statistically different from that in the positive control group ( P The result (>0.05) indicates that both drugs are comparable in their ability to downregulate the expression of fibrosis-related genes. These results suggest that *Gynostemma pentaphyllum* can inhibit the abnormal upregulation of liver fibrosis-related genes in MASH model mice, indicating that it has a regulatory effect on MASH-related liver fibrosis molecules.

[0064] Figure 4 A to Figure 4 The experimental data corresponding to C are shown in Table 4.

[0065] Table 4. In mouse liver Srebp1, FASN, SCD1, TNF-α, MCP-1, IL-1β, Col3α1, Col1α1, CTGF Summary table of relative gene expression levels

[0066] As shown in Table 4, for mice Srebp1 In terms of relative gene expression, the low-dose group of Gynostemma pentaphyllum showed an 84.5% reduction compared to the model group, the high-dose group showed a 99.6% reduction compared to the model group, and the positive control group showed a 99.7% reduction compared to the model group; in mice... FASN In terms of relative gene expression levels, the low-dose group of Gynostemma pentaphyllum showed an 89.1% reduction compared to the model group, the high-dose group showed a 99.0% reduction compared to the model group, and the positive control group showed a 98.3% reduction compared to the model group; in mice... SCD1 In terms of relative gene expression, the low-dose group of Gynostemma pentaphyllum showed a 59.4% decrease compared to the model group, the high-dose group showed a 99.4% decrease compared to the model group, and the positive control group showed a 98.6% decrease compared to the model group; in mice... TNF-α In terms of relative gene expression levels, the low-dose group of Ganmutong showed a 78.8% reduction compared to the model group, the high-dose group showed an 88.1% reduction compared to the model group, and the positive control group showed an 84.2% reduction compared to the model group; in mice... MCP-1 In terms of relative gene expression, the low-dose group of Ganmutong showed a 48.0% decrease compared to the model group, the high-dose group showed a 93.6% decrease compared to the model group, and the positive control group showed an 81.9% decrease compared to the model group; in mice... IL-1β In terms of relative gene expression levels, the low-dose group of Gynostemma pentaphyllum showed a 52.0% decrease compared to the model group, the high-dose group showed a 91.0% decrease compared to the model group, and the positive control group showed a 60.3% decrease compared to the model group; in mice... Col3α1 In terms of relative gene expression, the low-dose group of Ganmutong showed a 72.6% reduction compared to the model group, the high-dose group showed a 97.3% reduction compared to the model group, and the positive control group showed a 96.0% reduction compared to the model group; in mice... Col1α1 In terms of relative gene expression levels, the low-dose group of Gynostemma pentaphyllum showed an 89.0% reduction compared to the model group, the high-dose group showed a 99.8% reduction compared to the model group, and the positive control group showed a 99.8% reduction compared to the model group; in mice... CTGF In terms of relative gene expression, the low-dose Ganmutong group was 90.0% lower than the model group, the high-dose Ganmutong group was 99.0% lower than the model group, and the positive control group was 96.4% lower than the model group.

[0067] This indicates that after intervention with Gynostemma pentaphyllum, the abnormal elevation of lipid synthesis-related genes, inflammation-related genes, and fibrosis genes in the liver tissue of MASH model mice induced by AMLN diet was significantly downregulated. This demonstrates that Gynostemma pentaphyllum can improve liver pathological changes in MASH model mice from multiple aspects, including abnormal lipid metabolism, inflammatory response, and fibrosis-related molecular responses. Specifically, the regulatory effect of high-dose Gynostemma pentaphyllum on lipid synthesis-related genes, inflammation-related genes, and fibrosis genes is comparable to or better than that of the positive control drug (Resmetirom). These results further support the possibility of using Gynostemma pentaphyllum in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-related steatohepatitis.

[0068] Example 4: Inhibitory effect of Aristolochia debilis on the expression of F4 / 80 and IL-1β, immune inflammatory markers in the liver of MASH model mice. This study induced a MASH model using an AMLN diet (40% fat, 2% cholesterol, 22% fructose). After 20 weeks of administration, mice were euthanized according to animal welfare guidelines, and liver tissue was collected. The mouse liver tissue was fixed, embedded, and sectioned. Immunohistochemical staining (IHC) was used to detect the protein expression and tissue distribution of the macrophage marker F4 / 80 and the inflammatory factor IL-1β in the liver tissue. The macrophage marker F4 / 80 was used to evaluate macrophage infiltration in the liver tissue; IL-1β was used to evaluate the level of local inflammatory response in the liver.

[0069] like Figure 5As shown in Figure A, only a small number of F4 / 80 positive cells, mainly hepatic macrophages (Kupffer cells), were observed in the liver tissue of normal control mice. Compared with the normal control group, the number of F4 / 80 positive cells in the liver tissue of model mice was significantly increased, and they were mainly distributed in areas of fatty degeneration and around inflammatory foci, indicating that macrophage infiltration in the liver tissue of MASH model mice was significantly increased after AMLN diet induction, and the hepatic immune inflammatory response was activated. Compared with the model group, the number of F4 / 80 positive cells in the liver tissue of mice in the low-dose group, high-dose group, and positive control group was significantly reduced. Among them, the reduction in the number of F4 / 80 positive cells in the liver tissue of mice in the high-dose group was comparable to that in the positive control group. The above results indicate that Glycyrrhiza uralensis can reduce the number of F4 / 80 positive cells in the liver tissue of MASH model mice, suggesting that it has the effect of inhibiting abnormal macrophage infiltration and local immune inflammatory response in the liver.

[0070] like Figure 5 As shown in Figure B, IL-1β positive staining was weak in the liver tissue of normal control mice, with only a small amount of positive expression. Compared with the normal control group, the IL-1β positive staining intensity was significantly enhanced in the liver tissue of model mice, with positive cells widely distributed within the liver lobules and around the inflammatory foci, further indicating that after AMLN induction, the expression of the pro-inflammatory factor IL-1β protein in the liver tissue of MASH model mice increased, and the liver inflammation response was aggravated. Compared with the model group, the IL-1β positive staining intensity and the number of IL-1β positive cells were significantly weakened in the liver tissue of mice in the low-dose Ganmutong group, high-dose Ganmutong group, and positive control group. These results indicate that Ganmutong can reduce the protein expression level of IL-1β in the liver tissue of MASH model mice, suggesting that it has an inhibitory effect on MASH-related liver inflammation.

[0071] This indicates that after AMLN diet induction, the number of F4 / 80 positive cells in the liver tissue of the model group mice increased, accompanied by enhanced IL-1β protein expression, suggesting increased macrophage infiltration and significant activation of the immune inflammatory response in the liver tissue of MASH model mice. After intervention with Gynostemma pentaphyllum, the number of F4 / 80 positive cells decreased, and the intensity of IL-1β positive staining significantly weakened, indicating that Gynostemma pentaphyllum can inhibit the abnormal increase of immune inflammatory markers in the liver of MASH model mice and alleviate inflammation-related changes in the liver. These results further support the use of Gynostemma pentaphyllum in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-related steatohepatitis.

[0072] Example 5: Inhibitory effect of Aristolochia debilis on the expression of COL1α1 and α-SMA, proteins related to liver fibrosis, in MASH model mice. This study induced a MASH model using an AMLN diet (40% fat, 2% cholesterol, 22% fructose). After 20 weeks of administration, mice were euthanized according to animal welfare guidelines, and liver tissue was collected. Total protein was extracted from an appropriate amount of liver tissue, and the expression levels of fibrosis-related proteins Collagen type I alpha1 (COL1α1) and α-smooth muscle actin (α-SMA) in liver tissue were detected by Western blotting. COL1α1 is an extracellular matrix collagen formation-related protein, and its elevated expression indicates enhanced matrix response related to liver fibrosis; α-SMA is an important marker of hepatic stellate cell activation and fibrosis progression, and its elevated expression indicates enhanced activation of fibrosis-related cells in the liver.

[0073] like Figure 6 A to Figure 6 As shown in C, compared with the normal control group, the expression levels of α-SMA and COL1α1 proteins in the liver tissue of mice induced by AMLN diet were significantly increased ( P The value <0.0001 indicates that after AMLN dietary induction, the expression of fibrosis-related proteins in the liver tissue of MASH model mice was upregulated, and the molecular response related to liver fibrosis was significantly activated. Compared with the model group, the relative expression level of α-SMA protein in the liver tissue of mice in the low-dose Ganmutong group did not change significantly, but the expression level of COL1α1 protein was significantly reduced, suggesting that low-dose Ganmutong can inhibit the abnormal expression of some liver fibrosis-related proteins, but its regulatory effect on α-SMA protein expression is not obvious.

[0074] Compared with the model group, the relative expression levels of α-SMA and COL1α1 proteins in the liver tissue of mice in the high-dose Ganmutong group and the positive control group were significantly reduced. P <0.0001). Among them, the high-dose Ganmutong group showed a greater downregulation of α-SMA protein expression than the positive control group. Meanwhile, the downregulation trend of COL1α1 protein expression in the high-dose Ganmutong group was not statistically different from that in the positive control group, suggesting that both groups were comparable in their effect on downregulating COL1α1 protein expression. These results further suggest that high-dose Ganmutong can inhibit the abnormal expression of liver fibrosis-related molecules in MASH model mice at the protein level.

[0075] Figure 6 B and Figure 6 The experimental data corresponding to C are shown in Table 5.

[0076] Table 5 Summary of relative expression levels of α-SMA and COL1α1 proteins

[0077] As shown in Table 5, regarding the relative expression level of mouse α-SMA protein, the model group increased by 243.0% compared with the normal control group, the low-dose Ganmutong group showed no significant change compared with the model group, the high-dose Ganmutong group decreased by 93.9% compared with the model group, and the positive control group decreased by 78.7% compared with the model group. Regarding the relative expression level of mouse COL1α1 protein, the model group increased by 87.0% compared with the normal control group, the low-dose Ganmutong group decreased by 4.3% compared with the model group, the high-dose Ganmutong group decreased by 42.2% compared with the model group, and the positive control group decreased by 41.2% compared with the model group.

[0078] This indicates that after AMLN diet induction, the expression of α-SMA and COL1α1 proteins increased in the liver tissue of MASH model mice, suggesting that the molecular response related to liver fibrosis was activated in the model mice. After intervention with *Gynostemma pentaphyllum*, especially high-dose intervention, the expression of α-SMA and COL1α1 proteins showed a significant decreasing trend. Furthermore, the regulatory effect of high-dose *Gynostemma pentaphyllum* on the expression of α-SMA and COL1α1 proteins was comparable to or better than that of the positive control drug (Resmetirom), indicating that *Gynostemma pentaphyllum* can inhibit the abnormal upregulation of liver fibrosis-related proteins in MASH model mice. This result corroborates the results of fibrosis-related gene expression in Example 3, further supporting the use of *Gynostemma pentaphyllum* in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-related steatohepatitis and its liver fibrosis-related pathological changes.

[0079] Example 6: Regulatory effect of Aristolochia debilis on the expression of NF-κB and TGF-β / Smad signaling pathway proteins in the liver of MASH model mice This study induced a MASH model using an AMLN diet (40% fat, 2% cholesterol, 22% fructose). After 20 weeks of administration, mice were euthanized according to animal welfare guidelines, and liver tissue was collected. Total protein was extracted from an appropriate amount of liver tissue. Western blotting was used to detect the expression levels of key proteins in the inflammation-related signaling pathway NF-κB and the fibrosis-related signaling pathway TGF-β / Smad in mouse liver tissue. Specifically, NF-κB signaling pathway-related indicators included NF-κBp65 (p65), phosphorylated NF-κBp65 (p-p65), IκBα, and phosphorylated IκBα (p-IκBα); TGF-β / Smad signaling pathway-related indicators included Smad2 / 3 and phosphorylated Smad2 / 3 (p-Smad2 / 3).

[0080] like Figure 7 A to Figure 7 As shown in C, compared with the normal control group, the p-p65 / p65 and p-IκBα / IκBα ratios in the liver tissue of the model group mice were significantly increased ( P<0.0001 indicates that the phosphorylation levels of NF-κB p65 and IκBα in the liver tissue of MASH model mice increased after AMLN induction, suggesting that NF-κB signaling pathway-related inflammatory signals were in an activated state. Compared with the model group, the p-p65 / p65 and p-IκBα / IκBα ratios in the liver tissue of mice in the low-dose Ganmutong group, high-dose Ganmutong group, and positive control group were all significantly decreased; among them, the p-IκBα / IκBα ratio in the low-dose Ganmutong group and positive control group showed a decreasing trend compared with the model group, but the difference was not statistically significant, while the high-dose Ganmutong group was significantly lower than the model group ( P <0.0001), and close to the level of the normal control group. The above results indicate that Gynostemma pentaphyllum can reduce the abnormal phosphorylation level of NF-κB inflammatory signaling-related proteins in the liver tissue of MASH model mice. Among them, the downregulation effect of high-dose Gynostemma pentaphyllum on the phosphorylation level of p65 and IκBα is more obvious, suggesting that it has a regulatory role in the abnormal activation of the NF-κB inflammatory signaling pathway.

[0081] like Figure 7 As shown in D, compared with the normal control group, the p-Smad2 / 3 / Smad2 / 3 ratio in the liver tissue of the model group mice was significantly increased ( P <0.0001 indicates that after AMLN induction, the phosphorylation level (p-Smad2 / 3) of Smad2 / 3 downstream of the TGF-β / Smad signaling pathway increased in the liver tissue of MASH model mice, and fibrosis-related signaling molecule responses were abnormally altered. Compared with the model group, the p-Smad2 / 3 / Smad2 / 3 ratio in the liver tissue of mice in the low-dose group, high-dose group, and positive control group all showed a decreasing trend, with the high-dose group showing a greater decrease than the low-dose group, showing a dose-related decreasing trend; the positive control group also showed a significant reduction in this ratio ( P <0.0001). The above results indicate that Gynostemma pentaphyllum can regulate the abnormal phosphorylation level of Smad2 / 3 in the liver tissue of MASH model mice, suggesting that it has a regulatory role in the alteration of downstream fibrosis-related signaling molecules associated with the TGF-β / Smad pathway.

[0082] Figure 7 B to Figure 7 The experimental data corresponding to D are shown in Table 6.

[0083] Table 6 Summary of p-p65 / p65, p-IκBα / IκBα, and p-Smad2 / 3 / Smad2 / 3 ratios

[0084] As shown in Table 6, regarding the p-p65 / p65 ratio in mice, the model group increased by 57.0% compared to the normal control group, the low-dose Ganmutong group decreased by 45.2% compared to the model group, the high-dose Ganmutong group decreased by 57.3% compared to the model group, and the positive control group decreased by 21.0% compared to the model group. Regarding the p-IκBα / IκBα ratio in mice, the model group increased by 48.0% compared to the normal control group, the low-dose Ganmutong group decreased by 2.7% compared to the model group, the high-dose Ganmutong group decreased by 30.4% compared to the model group, and the positive control group decreased by 5.4% compared to the model group. Regarding the p-Smad2 / 3 / Smad2 / 3 ratio in mice, the model group increased by 57.0% compared to the normal control group, the low-dose Ganmutong group decreased by 45.9% compared to the model group, the high-dose Ganmutong group decreased by 58.0% compared to the model group, and the positive control group decreased by 71.3% compared to the model group.

[0085] This indicates that after AMLN diet induction, the p-p65 / p65 and p-IκBα / IκBα ratios in the liver tissue of the model group mice increased, as did the p-Smad2 / 3 / Smad2 / 3 ratio, suggesting that abnormal changes occurred in the NF-κB-related inflammatory signaling pathway and the TGF-β / Smad-related fibrosis signaling pathway in the liver tissue of MASH model mice. After intervention with Gynostemma pentaphyllum, the p-p65 / p65 and p-IκBα / IκBα ratios decreased, and the p-Smad2 / 3 / Smad2 / 3 ratio also decreased. This indicates that Gynostemma pentaphyllum can regulate abnormal changes in downstream proteins of the NF-κB and TGF-β / Smad signaling pathways in the liver tissue of MASH model mice, suggesting that it has an inhibitory effect on the abnormal activation of inflammation and fibrosis-related signals. Furthermore, low-dose and high-dose Gynostemma pentaphyllum showed better regulatory effects on the p-p65 / p65 ratio than the positive control drug (Resmetirom), and high-dose Gynostemma pentaphyllum showed better regulatory effects on the p-IκBα / IκBα ratio than the positive control drug (Resmetirom). Combined with the aforementioned liver histopathology, blood biochemistry, and expression results of inflammation and fibrosis-related genes, these results further support the use of Gynostemma pentaphyllum in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-related steatohepatitis and its inflammatory and fibrosis-related pathological changes.

[0086] The above examples demonstrate that: First, after intervention with Gynostemma pentaphyllum, especially high-dose Gynostemma pentaphyllum intervention, the body weight and liver weight of MASH model mice induced by AMLN diet were significantly reduced, and abnormal liver color and volume were significantly improved. Pathological changes in liver tissue, such as lipid deposition, inflammatory cell infiltration, ballooning degeneration, and collagen deposition, were significantly alleviated. Second, lipid metabolism indicators TC and TG in mouse plasma and liver tissue, as well as liver function indicators ALT and AST, were effectively improved, proving that Gynostemma pentaphyllum has an ameliorative effect on MASH-related liver lipid accumulation and liver function damage. Third, Gynostemma pentaphyllum can reduce... Srebp1, FASN, SCD1 Increasing the expression of lipid synthesis-related genes improves hepatic steatosis and reduces... TNF-α, MCP-1, IL-1β Inflammation-related genes and Col3α1, Col1α1, CTGF Fourth, Ganmutong can reduce the number of F4 / 80 positive cells and IL-1β protein expression in liver tissue, alleviate hepatic macrophage infiltration and inflammatory response, and downregulate the expression of α-SMA and COL1α1 fibrosis-related proteins, thus improving liver fibrosis-related changes. Fifth, Ganmutong can downregulate the ratio of p-p65 / p65 and p-IκBα / IκBα, which are related to the NF-κB inflammatory signaling pathway, and regulate the ratio of p-Smad2 / 3 / Smad2 / 3, which are downstream related proteins of the TGF-β / Smad fibrosis signaling pathway, suggesting that its ameliorative effect on MASH may be related to the inhibition of the activation of inflammatory signaling pathways and fibrosis signaling pathways. In conclusion, Ganmutong can alleviate and / or treat hepatic steatosis, inflammatory response, and fibrotic damage in MASH model animals induced by AMLN diet, thus providing potential for the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-related steatohepatitis.

[0087] Finally, it should be noted that the above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention should be considered equivalent substitutions and are included within the protection scope of the present invention.

Claims

1. Application of Gynostemma pentaphyllum in the preparation of drugs for the prevention and / or treatment of metabolic dysfunction-associated steatohepatitis (MASH).

2. The application according to claim 1, characterized in that, The drug has Aristolochia debilis as its main active ingredient.

3. The application according to claim 2, characterized in that, The drug has Gynostemma pentaphyllum as its sole active ingredient.

4. The application according to claim 2 or 3, characterized in that, The drug also includes pharmaceutically commonly used excipients or auxiliary ingredients.

5. The application according to claim 1, characterized in that, The Ganmutong mentioned is the original medicinal material of Ganmutong.

6. The application according to claim 1, characterized in that, The guanosine monosodium glutamate is guanosine monosodium glutamate powder.

7. The application according to claim 1, characterized in that, The drug is an oral formulation.

8. The application according to claim 7, characterized in that, The oral preparations include any one of tablets, capsules, granules, powders, oral liquids, or pills.

9. The application according to claim 1, characterized in that, The metabolic dysfunction-related steatohepatitis mentioned above is metabolic dysfunction-related steatohepatitis accompanied by liver fibrosis.

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