Traditional Chinese medicine compound preparation for treating non-alcoholic steatohepatitis and application thereof
By using traditional Chinese medicine compound preparations with ingredients such as ginkgo leaves, Panax notoginseng, Coptis chinensis, etc., the problem of lack of effective treatment of NASH in the prior art was solved, and the effect of significantly improving the clinical characteristics and biochemical indicators of the NASH mouse model was achieved.
Patent Information
- Application Number
- CN202510095754.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-22
- Publication Date
- 2025-04-11
AI Technical Summary
There is no effective treatment for non-alcoholic steatohepatitis (NASH) in the prior art, resulting in a lack of options for the treatment of the disease.
A Chinese medicine compound preparation, including Ginkgo leaves, Panax notoginseng, Coptis chinensis, Alisma, Mianma Guanzhong, Artemisia chen, Polygonum multiflorum, Curcuma Curcuma and Atractylodes macrocephala, was prepared by specific extraction methods to treat NASH.
This Chinese medicine compound preparation significantly improved the weight, liver appearance, liver index, serum biochemical index and liver histopathological changes in NASH mouse models, and improved insulin resistance, providing a new option for the treatment of NASH.
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Figure CN119792475A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of biomedicine, and in particular relates to a Chinese medicinal compound preparation for treating non-alcoholic fatty hepatitis and an application thereof. Background Art
[0002] Nonalcoholic Steatohepatitis (NASH) is a severe form of nonalcoholic fatty liver disease (NAFLD), with clinical manifestations of diffuse macrovesicular steatosis of hepatocytes. It is a key cause of advanced liver disease (such as cirrhosis) and has become the second most common indication for liver transplantation worldwide, seriously endangering people's lives and health. However, due to its complex pathogenesis, there are currently no marketed drugs for the treatment of NASH. Therefore, finding a suitable, more effective and safe drug to treat NASH is an urgent problem to be solved in clinical practice. Summary of the invention
[0003] The technical problem to be solved by the present invention is to provide a new option for patients with non-alcoholic fatty hepatitis.
[0004] The technical solution of the present invention is as follows: The invention provides a traditional Chinese medicine compound preparation for treating nonalcoholic fatty hepatitis, comprising the following ingredients: ginkgo leaf, notoginseng, coptis root, oriental water chestnut, cyperus rotundus, capillaris, polygonum cuspidatum, curcuma and atractylodes.
[0005] Furthermore, the dosage of each ingredient is as follows: 3-12 parts by weight of Ginkgo biloba leaves, 4.5-18 parts by weight of Panax notoginseng, 2.5-10 parts by weight of Coptis chinensis, 3-12 parts by weight of Alisma orientalis, 3-18 parts by weight of Cynomorium mongolicum, 3-12 parts by weight of Artemisia capillaris, 5-20 parts by weight of Polygonum cuspidatum, 3-18 parts by weight of Curcuma zedoaria and 5-20 parts by weight of Atractylodes macrocephala.
[0006] Preferably, the dosage of each ingredient is as follows: 6 parts by weight of Ginkgo biloba leaves, 9 parts by weight of Panax notoginseng, 5 parts by weight of Coptis chinensis, 6 parts by weight of Alisma orientalis, 9 parts by weight of Cynomorium miannanense, 6 parts by weight of Artemisia capillaris, 10 parts by weight of Polygonum cuspidatum, 9 parts by weight of Curcuma zedoaria and 10 parts by weight of Atractylodes macrocephala.
[0007] The present invention also provides a method for preparing the Chinese medicine compound preparation, comprising the following steps: a. Mix ginkgo leaves, Panax notoginseng, Coptis chinensis and Polygonum cuspidatum, extract twice with 70% ethanol as extract, combine the extracts, remove excess ethanol, and obtain filtrate 1; b. Mix the drug residue extracted in step a with Rhizoma Alismatis, Rhizoma Cynomorii, Artemisia Capillaris, Rhizoma Curcumae, and Rhizoma Atractylodis Macrocephalae, add water for extraction, extract twice, combine the extracts, and obtain filtrate 2; c. Combine filtrate 1 and filtrate 2, concentrate, dry and crush.
[0008] Furthermore, in both step a and step b, the extraction is carried out by a heating, condensing and reflux method; in step a, a rotary evaporator is used to remove excess ethanol.
[0009] Specifically, in step a, the liquid-to-material volume-to-mass ratio (mL / g) is 1:6 to 1:20.
[0010] Preferably, in step a, the liquid-to-material volume-to-mass ratio (mL / g) is 1:6.
[0011] Specifically, in step a, each extraction time is 1 to 3 hours.
[0012] Preferably, in step a, each extraction time is 1.5 h.
[0013] Specifically, in step b, the liquid-to-material volume-to-mass ratio (mL / g) is 1:6-1:12.
[0014] Preferably, in step b, the liquid-to-material volume-to-mass ratio (mL / g) is 1:8.
[0015] Specifically, in step b, each extraction time is 1 to 3 hours.
[0016] Preferably, in step b, each extraction time is 1.5 h.
[0017] The present invention also provides the use of the above-mentioned Chinese medicine compound preparation or the Chinese medicine compound preparation prepared by the above-mentioned method in preparing a medicine for treating or preventing non-alcoholic fatty liver disease.
[0018] The present invention also provides the use of the above-mentioned Chinese medicine compound preparation or the Chinese medicine compound preparation prepared by the above-mentioned method in the preparation of drugs for improving insulin resistance and glucose intolerance, controlling body weight and food intake, reducing body fat rate, reducing blood sugar, reducing liver damage, regulating lipid metabolism, improving fatty liver, improving blood lipids, improving inflammatory response, improving liver tissue oxidative stress level and / or improving liver steatosis, inflammation, fibrosis or liver damage caused by a high-fat and high-sugar diet.
[0019] The present invention also provides a medicine, food or health product for preventing or treating obesity, glucose and lipid metabolism disorders, and non-alcoholic fatty liver disease, including the above-mentioned Chinese medicine compound preparation or the Chinese medicine compound preparation prepared by the above-mentioned method.
[0020] Beneficial effects of the present invention: Based on clinical experience, the present invention formulates a Chinese medicine compound, which is as follows: Ginkgo biloba, Panax notoginseng, Coptis chinensis, Alisma orientalis, Rhizoma Cynomorii, Artemisia capillaris, Polygonum cuspidatum, Curcuma zedoaria and Atractylodes macrocephala, which has the efficacy of treating non-alcoholic fatty hepatitis. The present invention provides an application of a Chinese medicine compound preparation for treating non-alcoholic fatty hepatitis in the preparation of a liver disease treatment drug. Based on the efficacy of the Chinese medicine compound preparation in clinical treatment of NASH, a mouse NASH model is established by a high-fat and high-cholesterol diet, and the NASH model mice are treated with the Chinese medicine compound preparation of the present invention. The effects of the Chinese medicine compound preparation on the changes in the weight, liver appearance, liver index, serum biochemical index, liver tissue pathology of the NASH mouse model and the effects of the Chinese medicine compound preparation on the insulin resistance of the NASH model animal are analyzed. It is the first time to find that the compound preparation has a significant therapeutic effect on the NASH model of mice established by a high-fat and high-cholesterol diet. The present invention provides a new option for the treatment of non-alcoholic fatty hepatitis and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 Effects of Chinese herbal compound O on insulin resistance in NASH model mice. A: OGTT blood glucose curve; B: area under the OGTT curve, n=7-8; C: HOMA-IR; n=8-10. ##P<0.01 VS. CON group, ####P<0.0001 VS. CON group, *P<0.05 VS. HFHC group, **P<0.01 VS. HFHC group, ns indicates no statistical significance.
[0022] Figure 2 Effects of Chinese herbal compound O on body weight, fat-to-lean ratio, blood glucose, liver index, AST and ALT. A: Body weight change curve; B: Fat-to-lean ratio of mice; C: Blood glucose level; D: Liver index; E: Serum ALT level; F: Serum AST level. #P<0.05 VS. CON group, ##P<0.01 VS. CON group, ####P<0.0001 VS. CON group, *P<0.05 VS. HFHC group, **P<0.01 VS. HFHC group, ***P<0.001 VS. HFHC group, n=8-10, ns indicates no statistical significance.
[0023] Figure 3Effects of Chinese herbal compound O on liver and serum biochemical indexes in NASH mouse model. A: liver TC level; B: liver TG level; C: serum TC level; D: serum TG level; E: serum high-density lipoprotein level; F: serum low-density lipoprotein level, n=3. ##P<0.01 VS. CON group, ####P<0.0001 VS. CON group, *P<0.05 VS. HFHC group, **P<0.01 VS. HFHC group, ***P<0.001 VS. HFHC group, n=8-10, ns indicates no statistical significance.
[0024] Figure 4 Effects of Chinese herbal compound O on H&E and Oil Red O staining of liver in NASH mouse model. DETAILED DESCRIPTION
[0025] Traditional Chinese medicine compound O (the traditional Chinese medicine compound preparation of the present invention) is a clinical experience prescription. Based on its clinical efficacy in treating NASH, the present invention establishes a NASH model in mice through a high-fat and high-cholesterol diet to comprehensively evaluate its improving effect on hepatocyte fatty degeneration during the development of NASH.
[0026] The following examples are preferred embodiments of the present invention and further illustrate the present invention in detail, but do not limit the embodiments of the present invention in any way. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principle of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.
[0027] The experimental animals used in the following examples are: Male C57BL / 6 mice (weight 20-25 g, age 8 weeks) were housed in the SPF animal room of the Experimental Animal Center of Guangdong Pharmaceutical University (20-25°C, 65-70%, 12 h light-dark alternation), and the animals were given free access to water and standard feed.
[0028] The experimental materials used in the following examples are: After 1 week of adaptive feeding, the mice were fed with a high-fat, high-cholesterol diet to induce a non-alcoholic fatty liver disease mouse model: 10 mice were randomly selected as the control group (CON) and fed with ordinary feed, and the remaining 50 mice were fed with a high-fat, high-cholesterol diet. At the end of the 8th week of modeling, the mice were divided into the model group (HFHC), the low-dose group of Chinese medicine compound (OL), the medium-dose group (OM), the high-dose group (OH), and the atorvastatin group (ATV), with 10 mice in each group. The drug was administered for 8 weeks. The body weight of the mice was recorded every week, and the blood glucose of the mice was recorded every four weeks.
[0029] Example 1 Extraction and drying of medicine ① Prepare the raw materials: weigh 6g of Ginkgo biloba leaves, 9g of Panax notoginseng, 5g of Coptis chinensis, 6g of Alisma orientalis, 9g of Cynomorium mongolicum, 6g of Artemisia capillaris, 10g of Polygonum cuspidatum, 9g of Curcuma zedoaria, and 10g of Atractylodes macrocephala (3-12g of Ginkgo biloba leaves, 4.5-18g of Panax notoginseng, 2.5-10g of Coptis chinensis, 3-12g of Alisma orientalis, 3-18g of Cynomorium mongolicum, 3-12g of Artemisia capillaris, 5-20g of Polygonum cuspidatum, 3-18g of Curcuma zedoaria, and 5-20g of Atractylodes macrocephala are also within the effective range and can also exert their medicinal effects). Ginkgo biloba leaves, Panax notoginseng, Coptis chinensis, and Polygonum cuspidatum are first mixed, and then heated, condensed, and refluxed for extraction. The extraction conditions are 70% ethanol as the extract, the liquid-to-material volume-to-mass ratio (mL / g) is 1:6, and the extraction is performed twice, each extraction time is 1.5h, and the first and second extracts are combined after the extraction is completed, and then the excess ethanol is removed by rotary evaporator to obtain filtrate 1; ② Mix the drug residue extracted in step ① with orientalis, cyperus rotundus, capillaris, zedoaria, and atractylodes, add water for extraction, and extract under the conditions of liquid-to-solid volume-to-mass ratio (mL / g) of 1:8, heat condensation reflux extraction twice, each time for 1.5 h, combine the extracts to obtain filtrate 2; ③ Combine filtrate 1 and filtrate 2, concentrate, dry and crush to obtain.
[0030] Example 2 Effect of Traditional Chinese Medicine Compound O on Insulin Resistance in NASH Mouse Model The mice were given the drug for 8 weeks, and their body weight was recorded every week, and their blood sugar was recorded every four weeks. After the 7th week of administration, the glucose tolerance of the mice was tested: fasting for 6 hours, gavage with 1g / kg glucose, and the blood sugar level of the mice was tested at 0 min, 30 min, 60 min, 90 min, and 120 min after gavage. The blood sugar of the mice was tested after fasting for 6 hours before sampling, and the fasting serum insulin content of the mice was tested by blood sampling from the tail vein, and the insulin resistance index (HOMA-IR) was calculated.
[0031] like Figure 1 Compared with the CON group, the glucose tolerance, area under the insulin tolerance curve and insulin resistance index of the HFHC group were significantly increased; compared with the HFHC group, after the Chinese medicine compound O intervened in the NASH model mice, the glucose tolerance and insulin resistance index of each group of mice were significantly reduced, indicating that the Chinese medicine compound O can significantly improve the insulin resistance of the NASH mouse model.
[0032] Example 3 Effects of Chinese medicine compound O on clinical characteristics of NASH mouse model According to the experimental steps of the Nanjing Jiancheng Aspartate Aminotransferase (AST) and Alanine Aminotransferase (ALT) kit instructions, the activity of AST and ALT in serum was detected. The weight of mice was recorded using a weight scale, the weight of mouse liver was weighed using a metering scale, and the liver index was calculated. The fat and lean meat ratio of mice was detected using a live mouse component analyzer and the data was recorded. The fasting blood glucose value of the mouse tail tip was detected using a Roche blood glucose meter and the data was recorded.
[0033] like Figure 2 As shown, compared with the CON group, the body weight, fat to lean ratio, blood sugar, liver index, and serum ALT levels of mice in the HFHC group were significantly increased; compared with the HFHC group, the Chinese medicine compound O could significantly reduce the fat to lean ratio, hypoglycemia, liver index, serum ALT and serum AST levels of NASH mice after treatment; the effect of the Chinese medicine compound O was similar to that of the atorvastatin group (ATV), indicating that the Chinese medicine compound O has a significant therapeutic effect on NASH model mice.
[0034] Example 4 Effects of Chinese Herbal Compound O on Liver and Serum Biochemical Indexes in NASH Model Animals According to the instructions of Nanjing Jiancheng triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) kits, TG and TC levels in liver tissue and serum were detected; serum HDL-C and LDL-C levels were detected.
[0035] like Figure 3 Compared with the CON group, the HFHC group mice had significantly increased liver and serum TG and TC levels, and significantly increased serum LDL-C levels; compared with the HFHC group, the Chinese medicine compound O could significantly reduce the liver and serum TG and TC levels, and significantly reduce the serum LDL-C level after treating NASH model mice, indicating that the Chinese medicine compound O has a significant regulatory effect on lipid metabolism in NASH model mice.
[0036] Example 5 Effect of Traditional Chinese Medicine Compound O on Liver Pathology and Oil Red O Staining in NASH Mice After animal dissection and sampling, the fresh liver tissue was immersed in tissue fixative and fixed for 48 hours; it was dehydrated using gradient ethanol dehydration method and then immersed in wax, embedded in paraffin, placed on a paraffin slicer for sectioning and H&E staining.
[0037] The frozen sections were dehydrated by sucrose dehydration method, embedded with OTC embedding agent, and placed on a cryostat for sectioning. The sections were fixed in formaldehyde-calcium solution for 10 min, washed with 60% isopropanol, stained with Oil Red O staining solution for 30 min, separated with 60% isopropanol to background color, counterstained with hematoxylin, sealed with glycerol gelatin, and observed under a light microscope for Oil Red staining.
[0038] like Figure 4 The livers of mice in each group were stained with H&E and Oil Red O. The results showed that compared with the CON group, the liver tissues of the HFHC group animals showed liver microvesicles and macrovesicles fatty degeneration, hepatic cord disorder, inflammatory cell infiltration, and multiple lipid droplets in the hepatocytes. The Chinese medicine compound O was able to reduce liver microvesicles and fatty degeneration in a dose-dependent manner after treating NASH model mice, and had a significant therapeutic effect on the NASH model of mice induced by a high-fat and high-cholesterol diet.
Claims
1. A Chinese medicinal compound preparation for treating non-alcoholic fatty hepatitis, characterized in that: It includes the following ingredients: Ginkgo biloba leaves, Panax notoginseng, Coptis chinensis, Alisma orientalis, Cynomorium fortunei, Artemisia capillaris, Polygonum cuspidatum, Curcuma zedoaria and Atractylodes macrocephala.
2. The Chinese medicine compound preparation according to claim 1, characterized in that: The dosage of each ingredient is as follows: 3-12 parts by weight of Ginkgo biloba leaves, 4.5-18 parts by weight of Panax notoginseng, 2.5-10 parts by weight of Coptis chinensis, 3-12 parts by weight of Alisma orientalis, 3-18 parts by weight of Cynomorium mongolicum, 3-12 parts by weight of Artemisia capillaris, 5-20 parts by weight of Polygonum cuspidatum, 3-18 parts by weight of Curcuma zedoaria and 5-20 parts by weight of Atractylodes macrocephala.
3. The Chinese medicine compound preparation according to claim 2, characterized in that: The dosage of each ingredient is as follows: 6 parts by weight of Ginkgo biloba leaves, 9 parts by weight of Panax notoginseng, 5 parts by weight of Coptis chinensis, 6 parts by weight of Alisma orientalis, 9 parts by weight of Cynomorium miannanense, 6 parts by weight of Artemisia capillaris, 10 parts by weight of Polygonum cuspidatum, 9 parts by weight of Curcuma zedoaria and 10 parts by weight of Atractylodes macrocephala.
4. The method for preparing the Chinese medicine compound preparation according to any one of claims 1 to 3, characterized in that: The steps include: a. Mix ginkgo leaves, Panax notoginseng, Coptis chinensis and Polygonum cuspidatum, extract twice with 70% ethanol as extract, combine the extracts, remove excess ethanol, and obtain filtrate 1; b. Mix the drug residue extracted in step a with Rhizoma Alismatis, Rhizoma Cynomorii, Artemisia Capillaris, Rhizoma Curcumae, and Rhizoma Atractylodis Macrocephalae, add water for extraction, extract twice, combine the extracts, and obtain filtrate 2; c. Combine filtrate 1 and filtrate 2, concentrate, dry and crush.
5. The preparation method according to claim 4, characterized in that: In both step a and step b, the extraction is performed by a heating, condensing and reflux method; in step a, a rotary evaporator is used to remove excess ethanol.
6. The preparation method according to claim 4, characterized in that: In step a, the liquid-to-solid volume-to-mass ratio (mL / g) is 1:6-1:20, and the extraction time for each time is 1-3 hours; in step b, the liquid-to-solid volume-to-mass ratio (mL / g) is 1:6-1:12, and the extraction time for each time is 1-3 hours.
7. The preparation method according to claim 6, characterized in that: In step a, the liquid-to-solid volume-to-mass ratio (mL / g) is 1:6, and the extraction time for each extraction is 1.5 h; in step b, the liquid-to-solid volume-to-mass ratio (mL / g) is 1:8, and the extraction time for each extraction is 1.5 h.
8. Use of the Chinese herbal compound preparation according to any one of claims 1 to 3 or the Chinese herbal compound preparation prepared by the method according to any one of claims 4 to 7 in the preparation of a medicament for treating or preventing non-alcoholic fatty liver disease.
9. Use of the Chinese medicine compound preparation according to any one of claims 1 to 3 or the Chinese medicine compound preparation prepared by the method according to any one of claims 4 to 7 in the preparation of drugs for improving insulin resistance and glucose intolerance, controlling body weight and food intake, reducing body fat percentage, reducing blood sugar, reducing liver damage, regulating lipid metabolism, improving fatty liver, improving blood lipids, improving inflammatory response, improving oxidative stress level in liver tissue and / or improving hepatic steatosis, inflammation, fibrosis or liver damage caused by a high-fat and high-sugar diet.
10. A medicine, food or health product for preventing or treating obesity, glucose and lipid metabolism disorders, and non-alcoholic fatty liver disease, characterized in that: The invention comprises the Chinese medicine compound preparation according to any one of claims 1 to 3 or the Chinese medicine compound preparation prepared by the method according to any one of claims 4 to 7.