A traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease

Abstract

This invention provides a traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease (NAFLD), relating to the field of biomedical technology. The composition comprises *Panax japonicus*, *Pinellia ternata*, *Sargentodoxa cuneata*, *Poria cocos*, *Citrus reticulata* leaves, *Schisandra chinensis*, *Rehmannia glutinosa*, and *Glycyrrhiza uralensis*. This composition targets the pathogenesis of NAFLD characterized by spleen deficiency, phlegm and blood stasis, and qi and yin deficiency, employing a combined approach of tonifying and purging, addressing both the root cause and symptoms, and possessing the functions of invigorating qi and nourishing yin, resolving phlegm and promoting blood circulation, and soothing the liver and unblocking the meridians. The composition reduces TC, TG, LDL-C, ALT, and AST levels in NAFLD model mice, increases HDL-C levels, and decreases IL-6 and TNF-α levels in liver tissue, thereby improving liver tissue damage and reducing hepatic fat deposition, thus exerting a therapeutic effect on NAFLD. This invention provides a new drug option for treating or improving NAFLD.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease. Background Technology

[0002] Non-alcoholic fatty liver disease (NAFLD) is a chronic, progressive liver disease characterized by hepatic steatosis and liver damage, occurring without significant alcohol intake. NAFLD patients often present with elevated blood triglycerides and decreased high-density lipoprotein cholesterol, and are often accompanied by obesity and type 2 diabetes. With the increasing prevalence of obesity and type 2 diabetes, the incidence and prevalence of NAFLD are rising. Approximately 6% to 30% of NAFLD patients may progress from simple steatosis to severe non-alcoholic steatohepatitis (NASH). Furthermore, NAFLD, metabolic syndrome, and type 2 diabetes are mutually reinforcing, collectively promoting the development of atherosclerotic cardiovascular disease, chronic kidney disease, liver decompensation, and malignant tumors such as hepatocellular carcinoma.

[0003] Currently, the core foundation of NAFLD treatment is lifestyle intervention, including achieving at least 5%-10% weight loss through dietary adjustments and regular exercise. While lifestyle interventions are effective, long-term patient adherence is extremely poor, making it difficult to maintain weight loss, and the degree of improvement in liver inflammation and fibrosis varies significantly among individuals. For patients with cardiovascular risk, statins and angiotensin-converting enzyme inhibitors are commonly used to control blood lipids and blood pressure to reduce the risk of cardiovascular events, but these drugs have very limited effects on improving liver pathology itself. Currently, there is a severe lack of specific drugs for liver damage. Although investigational drugs such as thyroid hormone receptor beta agonists (Resmetirom) have been recommended by some guidelines for patients at specific stages in recent years, their high cost, limited applicable population, and lack of long-term data due to their short time on the market remain significant challenges.

[0004] Traditional Chinese medicine (TCM) possesses the characteristics of multi-target holistic regulation and mild medicinal properties, giving it unique advantages in the prevention and treatment of non-alcoholic fatty liver disease (NAFLD). TCM compound formulas are widely used clinically to treat fatty liver with good results. This is mainly achieved by strengthening the spleen and soothing the liver to regulate lipid metabolism at its root, resolving phlegm and dampness to clear accumulated fat in the liver, and simultaneously inhibiting hepatic oxidative stress and local inflammatory damage. This effectively reduces transaminase levels, alleviates the degree of hepatic steatosis, and slows the progression of liver fibrosis. This invention aims to provide a TCM composition with definite efficacy and high safety for the treatment or improvement of NAFLD. Summary of the Invention

[0005] (a) Technical problems to be solved

[0006] In view of the shortcomings of the prior art, the present invention provides a traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease.

[0007] (II) Technical Solution

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] A traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease, the composition comprising the following traditional Chinese medicinal materials in parts by weight: 10-30 parts of Panax notoginseng, 5-20 parts of Pinellia ternata, 5-25 parts of Sargentodoxa cuneata, 5-25 parts of Poria cocos, 5-20 parts of Citrus reticulata leaves, 5-20 parts of Schisandra chinensis, 5-25 parts of Rehmannia glutinosa, and 1-15 parts of Glycyrrhiza uralensis.

[0010] Preferably, the traditional Chinese medicine composition consists of the following traditional Chinese medicinal materials in parts by weight: 15-25 parts of Panax notoginseng, 5-15 parts of Pinellia ternata, 10-20 parts of Sargentodoxa cuneata, 10-20 parts of Poria cocos, 5-15 parts of Citrus reticulata leaves, 5-15 parts of Schisandra chinensis, 10-20 parts of Rehmannia glutinosa, and 5-10 parts of Glycyrrhiza uralensis.

[0011] Preferably, the traditional Chinese medicine composition consists of the following traditional Chinese medicinal materials in parts by weight: 20 parts of Panax notoginseng, 9 parts of Pinellia ternata, 15 parts of Sargentodoxa cuneata, 15 parts of Poria cocos, 6 parts of Citrus reticulata leaves, 6 parts of Schisandra chinensis, 12 parts of Rehmannia glutinosa, and 6 parts of Glycyrrhiza uralensis.

[0012] Furthermore, the traditional Chinese medicine composition provided by the present invention is used to prepare a drug for treating or improving non-alcoholic fatty liver disease. The drug is made from the above-mentioned parts by weight of traditional Chinese medicinal materials as raw materials, supplemented with pharmaceutically acceptable excipients, and prepared into any one of the following oral dosage forms: oral liquid, granules, suspension, powder, decoction, pills, tablets, and capsules.

[0013] The medicinal properties of each Chinese medicinal material in the herbal composition of this invention are described below:

[0014] Panax japonicus (also known as pearl ginseng) is the rhizome of Panax japonicus C. A. Mey. var. major (Burk.) CY Wu et KM Feng, a plant in the Araliaceae family. It has a bitter and sweet taste and is slightly cold in nature. It enters the liver, lung, and stomach meridians. Its functions include nourishing lung yin, removing blood stasis and relieving pain, and stopping bleeding. It is mainly used for deficiency of both qi and yin, feverish thirst, consumptive cough, hemoptysis, hematemesis, epistaxis, metrorrhagia, and traumatic bleeding.

[0015] Prepared Pinellia: A processed product of the tuber of Pinellia ternata (Thunb.) Breit., a plant in the Araceae family; pungent in taste and warm in nature. It enters the spleen, stomach, and lung meridians. Its function is to dry dampness and resolve phlegm. It is mainly used for cough and asthma with excessive phlegm, dizziness and palpitations due to phlegm retention, vertigo due to wind-phlegm, and headache due to phlegm syncope.

[0016] Poria cocos root: The pine root from the sclerotium of the fungus Poria cocos (family Polyporaceae); sweet in taste and neutral in nature. It enters the liver and heart meridians. Its function is to calm the liver and soothe the mind. It is mainly used for palpitations, forgetfulness, facial paralysis, muscle spasms due to wind-heat, aphasia, and mental disturbances.

[0017] Sargentodoxa cuneata (Oliv.) Rehd. et Wils., a plant belonging to the genus Sargentodoxa in the family Lardizabalaceae, is a bitter and neutral plant. It enters the liver and large intestine meridians. Its functions include clearing heat and detoxifying, promoting blood circulation, dispelling wind and relieving pain. It is mainly used for intestinal abscesses and abdominal pain, carbuncles and sores caused by heat toxins, amenorrhea, dysmenorrhea, traumatic swelling and pain, and rheumatic pain.

[0018] Schisandra chinensis: The dried, ripe fruit of Schisandra chinensis (Turcz.) Baill., a plant in the Magnoliaceae family; it has a sour and sweet taste and is warm in nature. It enters the lung, heart, and kidney meridians. Its functions include astringing and consolidating, replenishing qi and generating fluids, tonifying the kidneys and calming the mind. It is mainly used for chronic cough and wheezing, nocturnal emission and spermatorrhea, enuresis and frequent urination, chronic diarrhea, spontaneous sweating and night sweats, thirst due to fluid depletion, internal heat and thirst, palpitations and insomnia.

[0019] Orange leaves: The leaves of Citrus reticulata Blanco and its cultivated varieties, belonging to the Rutaceae family; bitter and pungent in taste, neutral in nature. It enters the Liver meridian. Its functions include soothing the liver, regulating qi, resolving phlegm, and reducing swelling and toxicity. It is mainly used for hypochondriac pain, mastitis, lung abscess, cough, chest tightness, and hernia.

[0020] Rehmannia glutinosa (raw): The dried tuberous root of Rehmannia glutinosa Libosch., a plant in the Scrophulariaceae family; sweet in taste and cold in nature. It enters the heart, liver, and kidney meridians. Its functions are to clear heat and cool the blood, nourish yin and generate fluids. It is mainly used for heat entering the blood level, febrile rashes, hematemesis and epistaxis, yin deficiency due to febrile diseases, red tongue with thirst, constipation due to fluid depletion, yin deficiency fever, steaming bone fever, and internal heat with thirst.

[0021] Prepared licorice root: A processed product of licorice root after being processed with honey; it is sweet in taste and neutral in nature. It enters the heart, lung, spleen, and stomach meridians. Its functions are to tonify the spleen and stomach, replenish qi, and restore the pulse. It is mainly used for spleen and stomach weakness, fatigue, palpitations, and irregular pulse.

[0022] (III) Beneficial Effects

[0023] This invention, based on the principles of traditional Chinese medicine (TCM) syndrome differentiation and treatment, comprehensively considers the patient's symptoms and etiology, and provides a TCM composition for treating or improving non-alcoholic fatty liver disease. The composition comprises *Panax japonicus*, *Pinellia ternata*, *Sargentodoxa cuneata*, *Poria cocos*, *Citrus reticulata* leaves, *Schisandra chinensis*, *Rehmannia glutinosa*, and *Glycyrrhiza uralensis*. *Panax japonicus* is the principal ingredient, chosen for its sweet, nourishing, and clearing properties. It effectively replenishes qi and yin, clears lung heat, and stops bleeding while promoting blood circulation. Its potent formula nourishes the qi of the middle jiao (spleen and stomach) and moistens the yin of the liver, ensuring that when qi is abundant, dampness and phlegm are eliminated, and when yin is sufficient, the liver meridians are nourished, thus removing blood stasis without harming the body's vital energy. The formula uses Pinellia ternata, Sargentodoxa cuneata, and Citrus reticulata leaves as assistant herbs. Pinellia ternata effectively dries dampness, resolves phlegm, and dissipates lumps and nodules, directly targeting the phlegm-dampness in the middle jiao, preventing dampness from accumulating and phlegm from forming. Sargentodoxa cuneata, bitter and neutral in nature, enters the blood level and effectively clears heat, detoxifies, invigorates blood, and unblocks the meridians, specifically resolving long-standing stagnant heat in the liver meridians and improving liver microcirculation to aid in the dissipation of lipoturbidity. Citrus reticulata leaves specifically target the qi level and effectively soothe the liver, promote qi circulation, dissipate nodules, and reduce swelling, clearing the key mechanism of liver and gallbladder stagnation, so that when qi flows smoothly, dampness is transformed and when qi flows freely, stasis is eliminated. The three herbs combined effectively eliminate the mutually binding evil of phlegm, stasis, and qi, assisting the principal herb in dispersing stasis and turbidity while ensuring that the elimination of evil does not harm the body's vital energy. Poria cocos, Schisandra chinensis, and Rehmannia glutinosa are added as adjuvant herbs. Poria cocos drains dampness, strengthens the spleen, and helps resolve phlegm, while also entering the heart and liver meridians to calm the mind, soothe the nerves, and relieve palpitations. Schisandra chinensis astringes lung qi and nourishes kidney yin, and its sourness helps to astringe the liver to prevent excessive dispersion. Rehmannia glutinosa, sweet and cold in nature, is moistening and effective in cooling the blood, nourishing yin, moistening dryness, and generating fluids. It can counteract the warming and drying effects of Pinellia ternata and Citrus reticulata leaves, assist the principal herb in nourishing yin and strengthening the body, and clear heat stagnation in the blood. These three herbs act as adjuvants, assisting the principal and assistant herbs in resolving phlegm, promoting blood circulation, and unblocking the meridians, while also replenishing qi, nourishing yin, and generating fluids, and compensating for yin and blood deficiency caused by liver meridian stagnation. Prepared licorice root, sweet and neutral, harmonizes the effects of the herbs, mitigating their harshness, and assists Ginseng in replenishing qi and harmonizing the middle jiao, making the overall formula more coordinated. This invention's herbal composition combines qi-tonifying and yin-nourishing herbs with phlegm-resolving and blood-activating herbs, simultaneously dispersing and tonifying, achieving the combined effects of replenishing qi and nourishing yin, resolving phlegm and activating blood, and soothing the liver and unblocking the meridians. This tonifies spleen deficiency, resolves phlegm and dampness, soothes liver stagnation, clears heat stagnation, and unblocks the meridians. It tonifies without stagnation and attacks without being drastic, treating both the root cause and the symptoms, thus achieving effective treatment for non-alcoholic fatty liver disease.

[0024] Based on the aforementioned TCM compatibility theory, this invention further explores and verifies the therapeutic effect of this TCM composition on non-alcoholic fatty liver disease (NAFLD). Experimental results show that the TCM composition of this invention reduces total cholesterol, triglycerides, low-density lipoprotein cholesterol, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels in NAFLD model mice, while increasing high-density lipoprotein cholesterol (HDL) levels, decreasing IL-6 and TNF-α levels in liver tissue, and improving liver tissue damage, thereby reducing hepatic fat deposition and exerting a therapeutic effect on NAFLD. This invention provides a new drug option for the treatment or improvement of NAFLD. Attached Figure Description

[0025] Figure 1 HE staining results of liver tissue from mice in each group.

[0026] Figure 2 Oil Red O staining results for liver tissues of mice in each group. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0028] Example 1

[0029] A traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease, the composition comprising the following traditional Chinese medicinal materials in parts by weight: 20 parts of Panax notoginseng, 9 parts of Pinellia ternata, 15 parts of Sargentodoxa cuneata, 15 parts of Poria cocos, 6 parts of Citrus reticulata leaves, 6 parts of Schisandra chinensis, 12 parts of Rehmannia glutinosa, and 6 parts of Glycyrrhiza uralensis.

[0030] The above-mentioned traditional Chinese medicine composition is used to prepare a drug for treating or improving non-alcoholic fatty liver disease. The drug is made from the above-mentioned parts by weight of traditional Chinese medicinal materials as raw materials, supplemented with pharmaceutically acceptable excipients, and prepared into any one of the following oral dosage forms: oral liquid, granules, suspension, powder, decoction, pills, tablets, and capsules.

[0031] Example 2

[0032] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition consists of the following parts by weight of traditional Chinese medicinal materials: 10 parts of Ginseng, 5 parts of Pinellia ternata, 5 parts of Sargentodoxa cuneata, 5 parts of Poria cocos, 5 parts of Citrus reticulata leaves, 5 parts of Schisandra chinensis, 5 parts of Rehmannia glutinosa, and 1 part of Glycyrrhiza uralensis.

[0033] Example 3

[0034] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition consists of the following parts by weight of traditional Chinese medicinal materials: 30 parts of Ginseng, 20 parts of Pinellia ternata, 25 parts of Sargentodoxa cuneata, 25 parts of Poria cocos, 20 parts of Citrus reticulata leaves, 20 parts of Schisandra chinensis, 25 parts of Rehmannia glutinosa, and 15 parts of Glycyrrhiza uralensis.

[0035] Example 4

[0036] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is composed of the following traditional Chinese medicine materials in parts by weight: 15 parts of Ginseng, 5 parts of Pinellia ternata, 10 parts of Sargentodoxa cuneata, 10 parts of Poria cocos, 5 parts of Citrus reticulata leaves, 5 parts of Schisandra chinensis, 10 parts of Rehmannia glutinosa, and 5 parts of Glycyrrhiza uralensis.

[0037] Example 5

[0038] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is composed of the following traditional Chinese medicine materials in parts by weight: 25 parts of Ginseng, 15 parts of Pinellia ternata, 20 parts of Sargentodoxa cuneata, 20 parts of Poria cocos, 15 parts of Citrus reticulata leaves, 15 parts of Schisandra chinensis, 20 parts of Rehmannia glutinosa, and 10 parts of Glycyrrhiza uralensis.

[0039] Example 6

[0040] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is composed of the following traditional Chinese medicine materials in parts by weight: 18 parts of Ginseng, 7 parts of Pinellia ternata, 12 parts of Sargentodoxa cuneata, 12 parts of Poria cocos, 9 parts of Citrus reticulata leaves, 9 parts of Schisandra chinensis, 14 parts of Rehmannia glutinosa, and 5 parts of Glycyrrhiza uralensis.

[0041] Example 7

[0042] The difference between this embodiment and Embodiment 1 is that the traditional Chinese medicine composition is composed of the following traditional Chinese medicine materials in parts by weight: 22 parts of Ginseng, 12 parts of Pinellia ternata, 18 parts of Sargentodoxa cuneata, 16 parts of Poria cocos, 12 parts of Citrus reticulata leaves, 9 parts of Schisandra chinensis, 12 parts of Rehmannia glutinosa, and 8 parts of Glycyrrhiza uralensis.

[0043] Experimental Example 1

[0044] Effects of the herbal composition of this invention on lipid deposition in a non-alcoholic fatty liver cell model

[0045] 1. Materials and Methods

[0046] 1.1 Drug Preparation

[0047] Experimental drug 1: 20g of *Panax notoginseng*, 9g of prepared *Pinellia ternata*, 15g of *Sargentodoxa cuneata*, 15g of *Poria cocos*, 6g of *Citrus reticulata* leaves, 6g of *Schisandra chinensis*, 12g of *Rehmannia glutinosa*, and 6g of prepared *Glycyrrhiza uralensis

[0048] Experimental drug 2: 9g of prepared Pinellia ternata, 15g of Sargentodoxa cuneata, 15g of Poria cocos, 6g of Citrus reticulata leaves, 6g of Schisandra chinensis, 12g of Rehmannia glutinosa, and 6g of prepared Glycyrrhiza uralensis.

[0049] Experimental drug 3: 20g of Panax notoginseng, 15g of Poria cocos, 6g of Schisandra chinensis, 12g of Rehmannia glutinosa, and 6g of Glycyrrhiza uralensis (processed).

[0050] Experimental drug 4: Gynostemma pentaphyllum 20g, Pinellia ternata 9g, Sargentodoxa cuneata 15g, Poria cocos 15g, Citrus reticulata leaf 6g, Schisandra chinensis 6g, Rehmannia glutinosa 12g, Glycyrrhiza uralensis 6g

[0051] Experimental drug 5: 20g of *Panax notoginseng*, 9g of dried tangerine peel, 15g of honeysuckle vine, 15g of *Poria cocos*, 6g of tangerine peel, 6g of schisandra fruit, 12g of *Rehmannia glutinosa*, and 6g of prepared licorice root.

[0052] Experimental drug 6: 20g of *Panax notoginseng*, 9g of *Pinellia ternata*, 15g of *Sargentodoxa cuneata*, 15g of *Poria cocos*, 6g of *Citrus reticulata* leaves, 6g of *Cornus officinalis*, 12g of *Scrophularia ningpoensis*, and 6g of *Glycyrrhiza uralensis* (processed).

[0053] Weigh each raw material according to the above weight ratio, each part by weight is 1g. After washing, soak in 8 times the amount of water for 30 minutes, then decoct for 30 minutes. Filter the decoction, add 4 times the amount of water and decoct for another 30 minutes. Combine the two decoctions and concentrate the drug content to about 1g / mL (equivalent to 1g of raw drug per mL). Store at 4℃ for later use. When using, prepare a drug-containing culture medium of the corresponding concentration.

[0054] 1.2 Cell Culture

[0055] Human L02 cells were selected as the research object. After the L02 cells were revived, they were seeded in RPMI 1640 medium containing 10% fetal bovine serum and 1% penicillin-streptomycin, and then cultured in a constant temperature incubator at 37°C and 5% CO2. When the cell confluence reached 80-90%, they were passaged or plated.

[0056] 1.3 Cell grouping, modeling and treatment

[0057] The optimal concentration of drugs 1-6 was determined to be 50 mg / mL based on previous experiments. Drugs 1-6 were then prepared into drug-containing culture media with a concentration of 50 mg / mL.

[0058] Logarithmic growth phase L02 cells were harvested, digested with trypsin, and then... (The sentence is incomplete and requires more context to translate accurately.) 5 Cells were seeded per well in 6-well plates and then divided into a blank control group, a model group, and drug groups 1-6, with 6 replicates in each group. Except for the blank control group, all other groups were induced with 2 mmol / L oleic acid for 24 h to establish a lipid precipitation model. Subsequently, each group was added with 200 μL of the corresponding drug-containing medium or blank medium and then incubated in a 37°C, 5% CO2 incubator for 24 h.

[0059] 1.4 Observation of lipid deposition in cells

[0060] Cells were collected 24 hours after treatment with different drugs, then washed with PBS solution and stained with Oil Red O. The formation of lipid droplets in each group of cells was observed under a microscope. The area of ​​lipid droplets in liver tissue was calculated using ImageJ, and the percentage of lipid droplet area was calculated.

[0061] Lipid droplet area percentage (%) = (Lipid droplet area / Total field of view area) × 100%

[0062] 1.5 Detection of triglyceride levels in cells

[0063] Cells were collected 24 hours after treatment with different drugs, and the triglyceride (TG) content in each group of cells was detected using a biochemical reagent kit.

[0064] 2 Results

[0065] As shown in Table 1, this experiment successfully constructed an in vitro NAFLD cell model using oleic acid induction. Compared with the control group, the proportion of lipid droplets and TG levels in the model group cells were significantly increased (P<0.01). After intervention with drugs 1-6, the proportion of lipid droplet area and TG levels in the NAFLD cell model decreased significantly to varying degrees (P<0.01). Among them, drug 1 (the traditional Chinese medicine composition of this invention) was significantly more effective than drugs 2-6 in reducing the proportion of lipid droplet area and TG levels in the NAFLD cell model (P<0.05 or P<0.01). The above results indicate that the medicinal ingredients in the traditional Chinese medicine composition of this invention, in a specific combination, complement each other and synergistically enhance each other to reduce lipid deposition and improve NAFLD. The absence of some medicinal ingredients or replacement of some drugs cannot achieve a good effect.

[0066] Table 1. Percentage of lipid droplets and TG levels in each group

[0067]

[0068] Note: Compared with the blank group, ** P<0.01; compared with the model group, ## P<0.01; compared with drug 1, & P<0.05, && P<0.01.

[0069] Experimental Example 2

[0070] 1. Materials and Methods

[0071] 1.1 Laboratory Animals

[0072] Six-week-old male Apoe- / - mice of SPF grade, weighing (22±2) g, were selected and housed in an environment with 20-25℃, 12h light-dark cycle, 50%-70% relative humidity, and sufficient food and water. The experiment began after 7 days of acclimatization feeding.

[0073] 1.2 Drugs and Preparation

[0074] The herbal extract of this invention consists of 20 parts of Panax notoginseng, 9 parts of Pinellia ternata, 15 parts of Sargentodoxa cuneata, 15 parts of Poria cocos, 6 parts of Citrus reticulata leaves, 6 parts of Schisandra chinensis, 12 parts of Rehmannia glutinosa, and 6 parts of Glycyrrhiza uralensis. Each part by weight is 1g. Weigh each raw material according to the above parts by weight, wash them, soak them in 8 times the amount of water for 30 minutes, then decoct for 30 minutes, filter the decoction, add 4 times the amount of water and decoct for another 30 minutes, combine the two decoctions, concentrate the drug content to about 1.27g / mL (equivalent to 1.27g of raw drug per mL), and store at 4℃ for later use.

[0075] Western medicine reference drug: Rosuvastatin tablets, 10mg / tablet, prepared with distilled water to form a 0.13mg / mL suspension, to be used immediately after preparation.

[0076] 1.3 Construction of a mouse model of non-alcoholic fatty liver disease

[0077] The control group mice were given a normal diet, while the other groups of mice were fed a high-fat diet (high-fat diet with 4.7 kcal / g calories, fat accounting for 40%-45% of calories, and cholesterol 0.15%) for a total of 6 weeks.

[0078] 1.4 Grouping and Administration

[0079] Forty Apoe- / - mice were randomly divided into four groups (n=8 per group): a blank control group, a model group, a positive control group, a low-dose group (hereinafter referred to as the low-dose group), and a high-dose group (hereinafter referred to as the high-dose group) of the traditional Chinese medicine composition of this invention. All mice were fed a high-fat diet and administered the medicine via gavage for 6 weeks. The positive control group received rosuvastatin suspension at 1.3 mg / kg, the low-dose group received the original traditional Chinese medicine solution at 1.27 g / kg, and the high-dose group received the original traditional Chinese medicine solution at 2.54 g / kg. The blank control group and the model group received 0.1 mL / 10 g of sterile water via gavage. All groups received the medicine once daily.

[0080] 1.5 Observation Indicators and Detection Methods

[0081] 1.5.1 Biochemical Detection

[0082] After the last administration, mice in each group were anesthetized with 0.5% sodium pentobarbital, and blood was collected from the heart and anticoagulated. Plasma was prepared by centrifugation. The levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) in the plasma of each group of mice were detected using biochemical reagent kits.

[0083] 1.5.2 Liver function tests

[0084] Mouse plasma was prepared according to the method in 1.5.1, and the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the plasma of each group of mice were detected using an automated biochemical analyzer.

[0085] 1.5.3 ELISA Detection

[0086] Liver tissue was homogenized with PBS at low temperature, and the levels of TNF-α and IL-6 in the mouse liver tissue homogenate were measured using an ELISA kit.

[0087] 1.5.4 HE staining

[0088] Mouse liver tissue was collected on ice. A portion of the mouse liver tissue was fixed in 4% paraformaldehyde for 24 h, washed, dehydrated, and embedded to prepare 5 μm thick paraffin sections. The sections were stained with hematoxylin and eosin sequentially, and the pathological changes in the liver tissue were observed under a microscope. The non-alcoholic fatty liver disease activity score (NAS) was used for evaluation, specifically based on steatosis (0-3 points), liver inflammation (0-3 points), and hepatocyte ballooning degeneration (0-2 points).

[0089] 1.5.5 Oil Red O staining

[0090] Mouse liver tissue was collected on ice, and a portion of the liver tissue was flash-frozen in liquid nitrogen to prepare 10 μm thick frozen sections. The sections were stained with 0.5% Oil Red O staining solution and counterstained with hematoxylin. The formation of lipid droplets in the liver tissue was observed under a microscope, and the area of ​​lipid droplets in the liver tissue was calculated using ImageJ. The area of ​​lipid droplets in the blank control group was normalized, and the relative area of ​​lipid droplet formation in each group of mice was calculated.

[0091] 1.6 Statistical Analysis

[0092] Statistical analysis was performed using IBM SPSS Statistics 19.0 software. One-way ANOVA was used for comparisons between groups. If the variances were homogeneous, the LSD test was used; if the variances were unequal, the Gameshoer test was used. Results are expressed as mean ± standard deviation. Format. P < 0.05 is considered statistically significant.

[0093] 2 Results

[0094] 2.1 Effects of the herbal composition of the present invention on biochemical indicators in mice with non-alcoholic fatty liver disease

[0095] As shown in Table 2, compared with the blank control group, the levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol in the plasma of mice in the model group were significantly increased, while the level of high-density lipoprotein cholesterol was significantly decreased (P<0.05). Compared with the model group, the levels of total cholesterol, triglycerides, and low-density lipoprotein cholesterol in the plasma of mice in the positive control group, low-dose group, and high-dose group were significantly decreased, while the level of high-density lipoprotein cholesterol was significantly increased (P<0.05).

[0096] Table 2. Biochemical indicators of mice in each group

[0097]

[0098] Note: # Compared with the model group, P<0.05; * Compared with the sham surgery group, P<0.05.

[0099] 2.2 Effects of the herbal composition of the present invention on liver function in mice with non-alcoholic fatty liver disease

[0100] Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are important indicators of liver function. As shown in Table 3, compared with the blank control group, the plasma ALT and AST levels of mice in the model group were significantly increased (P<0.05); compared with the model group, the plasma ALT and AST levels of mice in the positive control group, low-dose group, and high-dose group were significantly decreased (P<0.05).

[0101] Table 3 Plasma ALT and AST levels in mice of each group

[0102]

[0103] Note: # Compared with the model group, P<0.05; * Compared with the sham surgery group, P<0.05.

[0104] 2.3 Effects of the herbal composition of the present invention on liver inflammation in mice with non-alcoholic fatty liver disease

[0105] Chronic inflammation is considered a key driver of NAFLD progression. Hepatic lipid accumulation activates immune cells, promoting the release of pro-inflammatory cytokines such as TNF-α and IL-6, thereby establishing a persistent inflammatory microenvironment. As shown in Table 4, compared with the blank control group, the levels of inflammatory markers TNF-α and IL-6 in the liver homogenate of mice in the model group were significantly increased (P<0.05); compared with the model group, the levels of inflammatory markers TNF-α and IL-6 in the liver homogenate of mice in the positive control group, low-dose group, and high-dose group were all significantly decreased (P<0.05).

[0106] Table 4. TNF-α and IL-6 content in liver homogenates of mice in each group

[0107]

[0108] Note: # Compared with the model group, P<0.05; * Compared with the sham surgery group, P<0.05.

[0109] 2.4 Effects of the herbal composition of the present invention on liver pathological changes in mice with non-alcoholic fatty liver disease

[0110] like Figure 1As shown, compared with the blank control group, the model group mice exhibited significant pathological changes in their livers, including severe hepatic steatosis, ballooning degeneration of cells, and inflammatory cell infiltration. Compared with the model group, the pathological changes in the livers of the positive control group, low-dose group, and high-dose group were all alleviated. The NAS scoring system was used to evaluate the pathological changes in the livers of each group of mice, as shown in Table 5. The results showed that compared with the blank control group, the score of the model group was significantly lower (P<0.05), while compared with the model group, the scores of the positive control group, low-dose group, and high-dose group were all significantly higher (P<0.05).

[0111] Table 5. Liver pathological scores of mice in each group.

[0112]

[0113] Note: # Compared with the model group, P<0.05; * Compared with the sham surgery group, P<0.05.

[0114] 2.5 Effect of the herbal composition of the present invention on lipid droplet formation in the liver of mice with non-alcoholic fatty liver disease

[0115] like Figure 2 As shown in Table 6, compared with the blank control group, obvious lipid droplet formation was observed in the liver of mice in the model group (P<0.05), while the lipid droplet area was significantly reduced in the positive control group, low-dose group and high-dose group compared with the model group (P<0.05).

[0116] Table 6. Lipid droplet formation in the liver of mice in each group.

[0117]

[0118] Note: # Compared with the model group, P<0.05; * Compared with the sham surgery group, P<0.05.

[0119] 3. Conclusion

[0120] Based on the above results, the herbal composition of this invention can significantly reduce total cholesterol, triglycerides, and low-density lipoprotein cholesterol levels in mice with non-alcoholic fatty liver disease (NAFLD), while increasing high-density lipoprotein cholesterol levels. It can also significantly reduce the levels of alanine aminotransferase and aspartate aminotransferase in the plasma of NAFLD mice, and decrease the levels of TNF-α and IL-6 in liver tissue. Simultaneously, it can significantly improve the pathological changes in the liver of NAFLD mice and reduce the formation of lipid droplets in the liver. These results indicate that the herbal composition of this invention has a significant therapeutic effect on NAFLD mice and can serve as a potential drug for treating or improving NAFLD.

[0121] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A traditional Chinese medicine composition for treating or ameliorating nonalcoholic fatty liver disease, characterized in that, The traditional Chinese medicine composition consists of the following parts by weight of Chinese medicinal materials: 10-30 parts of Panax notoginseng, 5-20 parts of Pinellia ternata, 5-25 parts of Sargentodoxa cuneata, 5-25 parts of Poria cocos, 5-20 parts of Citrus reticulata leaves, 5-20 parts of Schisandra chinensis, 5-25 parts of Rehmannia glutinosa, and 1-15 parts of Glycyrrhiza uralensis. 2.The traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease according to claim 1, wherein, The traditional Chinese medicine composition consists of the following parts by weight of Chinese medicinal materials: 15-25 parts of Panax notoginseng, 5-15 parts of Pinellia ternata, 10-20 parts of Sargentodoxa cuneata, 10-20 parts of Poria cocos, 5-15 parts of Citrus reticulata leaves, 5-15 parts of Schisandra chinensis, 10-20 parts of Rehmannia glutinosa, and 5-10 parts of Glycyrrhiza uralensis. 3.The traditional Chinese medicine composition for treating or improving non-alcoholic fatty liver disease according to claim 1, wherein, The traditional Chinese medicine composition consists of the following parts by weight of Chinese medicinal materials: 20 parts of Gynostemma pentaphyllum, 9 parts of Pinellia ternata, 15 parts of Sargentodoxa cuneata, 15 parts of Poria cocos, 6 parts of Citrus reticulata leaves, 6 parts of Schisandra chinensis, 12 parts of Rehmannia glutinosa, and 6 parts of Glycyrrhiza uralensis.

4. The use of the traditional Chinese medicine composition according to any one of claims 1 to 3 in the preparation of a drug for treating or improving non-alcoholic fatty liver disease, characterized in that, The drug is made from the above-mentioned proportions of Chinese medicinal herbs by weight, supplemented with pharmaceutically acceptable excipients, and prepared into any one of the following oral dosage forms: oral liquid, granules, suspension, powder, decoction, pills, tablets, and capsules.

Images