A traditional Chinese medicine composition, a preparation method therefor and application thereof

Abstract

This invention belongs to the technical field of traditional Chinese medicine compositions, specifically relating to a traditional Chinese medicine composition, its preparation method, and its application. By weight, the raw materials of the traditional Chinese medicine composition include: 1-31 parts of Gynostemma pentaphyllum, 6-50 parts of Ganoderma lucidum, 12-40 parts of Paeonia lactiflora, 6-34 parts of Citrus aurantium, and 4-28 parts of Allium macrostemon, preferably 10-12 parts of Gynostemma pentaphyllum, 22-28 parts of Ganoderma lucidum, 25-28 parts of Paeonia lactiflora, 15-22 parts of Citrus aurantium, and 18-22 parts of Allium macrostemon. The preparation method of the traditional Chinese medicine composition of this invention ensures the transfer rate and effectiveness of the active ingredients of each raw material while maintaining a low preparation cost. The traditional Chinese medicine composition of this invention can effectively improve atherosclerosis and hyperlipidemia, producing a synergistic effect.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine composition technology, specifically relating to a traditional Chinese medicine composition, its preparation method, and its application. Background Technology

[0002] Atherosclerosis (AS) is one of the most important pathological bases of cardiovascular disease, a major cause of death from myocardial infarction, cerebral infarction, and coronary heart disease, and is related to the development of many other diseases. The main pathological feature of atherosclerosis is the deposition of lipids in the subendothelial layer of certain parts of the artery, accompanied by the proliferation of smooth muscle cells and fibrous components. This gradually develops into localized plaques, causing the arterial wall to thicken and harden. Necrosis and disintegration of the plaque tissue combine with the deposited lipids, forming the pathological changes of atherosclerotic material. Cholesterol and cholesterol esters are the main components of atherosclerotic plaques. When cholesterol levels are too high, especially oxidized low-density lipoprotein (OX-LDL) and triglycerides, it can be phagocytosed by macrophages and deposited under the arterial endothelium, causing subendothelial degeneration. This leads to platelet aggregation on the arterial wall. If arterial wall damage or cholesterol transport disorders are present, lipid plaques are easily formed in the arterial intima. Finally, the vessel wall bulges and protrudes into the lumen, forming atherosclerotic plaques.

[0003] Lipoprotein lipase (LPL) is a key enzyme in lipid metabolism. It is synthesized and secreted into the blood by extrahepatic parenchymal cells to hydrolyze chylomicrons (CM) and triglycerides (TG) on VLDL. LPL deficiency will cause increased levels of CM and VLDL.

[0004] For the treatment of atherosclerosis (AS), lipid-lowering strategies are the most basic treatment for hyperlipidemia. Statins are commonly used and can effectively lower blood lipid levels, slow the progression of AS, stabilize coronary artery plaques, and reduce LDL cholesterol levels. However, studies have shown that the vascular protective function of statins has certain limitations. Some patients taking statins still have a risk of cardiovascular disease. Moreover, high-dose statins can also produce side effects, mainly hepatotoxicity and muscle toxicity, including elevated liver enzymes, muscle pain, and even rhabdomyolysis. Long-term use of statins can cause liver damage.

[0005] Chinese invention patent application CN201210136593.5 discloses a traditional Chinese medicine for treating atherosclerosis and its preparation method, belonging to the field of traditional Chinese medicine preparation technology. It is made from the following raw materials in the indicated weight proportions: 10g-20g of Citrus aurantium, 8g-15g of Bambusa textilis, 10g-20g of Citrus reticulata peel, 10g-30g of Poria cocos, 8g-15g of Acorus tatarinowii, 8g-15g of Curcuma longa, 20g-40g of Trichosanthes kirilowii, 10g-20g of Allium macrostemon, 10g-20g of Pyrrosia lingua, 20g-40g of Spatholobus suberectus, 0.5g-2g of Panax notoginseng, and 0.5g-2g of Panax quinquefolius. The preparation steps are as follows: 1. Grind the Panax notoginseng and American ginseng from the above-mentioned Chinese herbs into powder and pass them through a 120-mesh sieve; 2. Add the remaining Chinese herbs—Citrus aurantium, Bambusa textilis, Citrus reticulata peel, Poria cocos, Acorus tatarinowii, Curcuma longa, Trichosanthes kirilowii, Allium macrostemon, Pyrrosia lingua, and Spatholobus suberectus—to 400-500 ml of water, soak for 30 minutes, and decoct for 15-20 minutes to obtain 100 ml of decoction; 3. Before drinking, add the Panax notoginseng powder and American ginseng powder to the above decoction according to their weight proportions, stir well, and drink. However, this involves a large number of medicinal materials, and the therapeutic effect of the drug composition in this application is unclear.

[0006] Chinese invention patent application CN201710616972.7 discloses a traditional Chinese medicine composition for the prevention and treatment of atherosclerosis and its preparation method. The composition consists of the following components in parts by weight: 2-8 parts of Acorus tatarinowii, 0.5-1.5 parts of deer antler glue, 1-5 parts of Trichosanthes kirilowii, 1-3 parts of Angelica dahurica, 2-6 parts of Saposhnikovia divaricata, 2-8 parts of Spatholobus suberectus, 0.5-1.5 parts of processed pangolin scales, 1-3 parts of Paeonia lactiflora, 2-8 parts of Forsythia suspensa, 2-8 parts of Scrophularia ningpoensis, 2-8 parts of Achyranthes bidentata, 1-3 parts of Asarum heterotropoides, 2-8 parts of Polygonum multiflorum, 5-10 parts of Astragalus membranaceus, 1-3 parts of Cinnamomum cassia, 1-3 parts of Polygonatum sibiricum, 2-6 parts of Fritillaria thunbergii, 1-3 parts of Prunella vulgaris, 1-3 parts of Gleditsia sinensis thorns, 2-8 parts of Trichosanthes kirilowii, and 1-3 parts of Allium macrostemon. This composition has a significant therapeutic effect on patients with atherosclerosis, but it requires some rare and scarce plants and animals, and is not suitable for widespread use.

[0007] Currently, there are many traditional Chinese medicines for the prevention and treatment of atherosclerosis and hyperlipidemia, but they still have shortcomings such as excessively large prescriptions, unclear efficacy, or high prices due to the use of expensive medicinal materials. Therefore, it is necessary to develop a low-cost and effective traditional Chinese medicine composition that can lower blood lipids and prevent and treat atherosclerosis. Summary of the Invention

[0008] To address the problems of high cost and large prescriptions in the existing technologies, this invention provides a traditional Chinese medicine composition, its preparation method, and its application. The traditional Chinese medicine composition provided by this invention has appropriate formulation and can effectively lower blood lipids and prevent atherosclerosis.

[0009] To achieve the above objectives, the present invention provides the following technical solution:

[0010] A traditional Chinese medicine composition, by weight, comprises the following raw materials: 1-31 parts of Gynostemma pentaphyllum, 6-50 parts of Ganoderma lucidum, 12-40 parts of Paeonia lactiflora, 6-34 parts of Citrus aurantium, and 4-28 parts of Allium macrostemon.

[0011] Preferably, the raw materials of the traditional Chinese medicine composition, by weight, include: 5-31 parts of Gynostemma pentaphyllum, 13-47 parts of Ganoderma lucidum, 12-30 parts of Paeonia lactiflora, 6-28 parts of Citrus aurantium and 14-26 parts of Allium macrostemon.

[0012] Preferably, the raw materials of the traditional Chinese medicine composition, by weight, include: 10-12 parts of Gynostemma pentaphyllum, 22-28 parts of Ganoderma lucidum, 25-28 parts of Paeonia lactiflora, 15-22 parts of Citrus aurantium and 18-22 parts of Allium macrostemon.

[0013] This invention also relates to a method for preparing the above-mentioned traditional Chinese medicine composition, comprising the following steps:

[0014] (1) Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon powder were extracted with water to obtain extract 1, which was filtered, concentrated and dried to obtain dried product 1;

[0015] (2) Extract Ganoderma lucidum and Paeonia lactiflora with alcohol to obtain extract 2, filter, concentrate and dry to obtain dried product 2;

[0016] (3) Mix dried substance 1 and dried substance 2 to obtain the product.

[0017] Preferably, the water extraction step in step (1) includes: adding 10-20 times the volume of water to the powders of Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon, extracting 1-3 times, each extraction lasting 1-3 hours, and combining the extracts to obtain extract 1; the concentration temperature is 50-70℃, and the concentration is carried out to a relative density of 1.01-1.25.

[0018] Preferably, the alcohol extraction step in step (2) includes: adding 12-18 times the volume of ethanol solution with a mass concentration of 30%-80% to the powder of Ganoderma lucidum and Paeonia lactiflora, extracting 1-3 times, each extraction for 1-3 hours, and combining the extracts to obtain extract 2; the concentration temperature is 50-70℃, and the concentration is carried out to a relative density of 1.01-1.25.

[0019] More preferably, the water extraction in step (1) is performed twice. The first extraction is performed by adding 14-16 times the volume of water and extracting for 1.3-1.7 hours. The second extraction is performed by adding 11-13 times the volume of water and extracting for 0.8-1.2 hours. The concentration temperature is 55-65°C, and the product is concentrated to a relative density of 1.01-1.15.

[0020] More preferably, the alcohol extraction in step (2) is performed twice. The first extraction is performed by adding 15-17 times the volume of an ethanol solution with a mass concentration of 40%-50% for 1.8-2.2 hours. The second extraction is performed by adding 11-13 times the volume of an ethanol solution with a mass concentration of 65%-75% for 1.3-1.7 hours. The concentration temperature is 55-65°C, and the product is concentrated to a relative density of 1.01-1.15.

[0021] The present invention also relates to the use of the above-described composition in the preparation of medicaments for the prevention and / or treatment of hyperlipidemia and its complications.

[0022] Preferably, the dosage form of the drug is selected from any one of tablets, granules, powders, and capsules; the drug is used to lower blood lipids, improve lipid metabolism disorders, or to prevent and / or treat atherosclerosis.

[0023] The present invention also relates to the application of the above-mentioned traditional Chinese medicine composition in the preparation of health products that help lower blood lipids.

[0024] The present invention also relates to pharmaceuticals or health products comprising the above-mentioned traditional Chinese medicine composition.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] (1) In the traditional Chinese medicine composition of the present invention, Gynostemma pentaphyllum can significantly reduce blood lipids and improve lipid metabolism disorders, thereby reducing the occurrence of atherosclerosis; Paeonia lactiflora can prevent liver oxidative damage and inhibit the upregulation of pro-inflammatory mediators, thus playing a protective role against ischemic brain injury. Ganoderma lucidum can improve the body's immune function, strengthen the body's homeostatic regulation ability, improve microcirculation, and promote the efficacy of the principal drug. Allium macrostemon can promote yang and dissipate stagnation, and regulate qi and relieve stagnation. Citrus aurantium has a strong qi-regulating effect and is good at regulating qi, which can guide the efficacy of the whole prescription to the whole blood vessels and effectively exert the function of lowering blood lipids. The results of the examples show that the traditional Chinese medicine preparation provided by the present invention can provide the effect of lowering blood lipids and can also improve atherosclerosis.

[0027] (2) The method for preparing the traditional Chinese medicine composition of the present invention ensures the transfer rate of the active ingredients of each raw material while keeping the preparation cost within a low range.

[0028] (3) The traditional Chinese medicine composition of the present invention can effectively improve atherosclerosis and produce a synergistic effect. Attached Figure Description

[0029] Figure 1 This is a comparative graph showing the effect of different doses of the traditional Chinese medicine composition on the body weight of mice in Example 1, compared with the high-fat diet group. * P<0.05, *** P<0.001;

[0030] Figure 2 This is a comparative graph showing the effects of different doses of the traditional Chinese medicine composition on liver lipid metabolism in Example 1;

[0031] Figure 3 This is a comparative graph showing the effects of different doses of the traditional Chinese medicine composition on blood lipids in Example 1, compared to the high-fat diet group. * P<0.05, ** P<0.01, *** P<0.001;

[0032] Figure 4 This is a comparative graph showing the effect of different dosages of the traditional Chinese medicine composition on oxidative stress in Example 1, compared to the high-fat diet group. **** P<0.0001;

[0033] Figure 5 This is a comparative graph showing the effects of different doses of the traditional Chinese medicine composition on the entire aortic plaque in Example 1, compared with the high-fat diet group. * P<0.05, ** P<0.01, **** P<0.0001;

[0034] Figure 6 This is a comparative graph showing the effect of different doses of the traditional Chinese medicine composition in Example 1 on the area of ​​plaque in the aortic root, compared with the high-fat diet group. **** P<0.0001;

[0035] Figure 7 This is a comparative graph showing the effect of different doses of the traditional Chinese medicine composition in Example 1 on the stability of aortic root plaques, compared with the high-fat diet group. **** P<0.0001;

[0036] Figure 8 This is a comparative graph showing the effect of different doses of the traditional Chinese medicine composition on the content of foam cells in plaques in Example 1, compared with the high-fat diet group. * P<0.05, ** P<0.01, **** P<0.0001;

[0037] Figure 9 This is a comparative graph showing the effect of different doses of the traditional Chinese medicine composition in Example 1 on the content of smooth muscle cells in plaques, compared with the high-fat diet group. ** P<0.01, *** P<0.001, **** P<0.0001;

[0038] Figure 10 This is a comparative graph showing the effect of different doses of the traditional Chinese medicine composition on the degree of plaque calcification in Example 1, compared with the high-fat diet group. ** P<0.01, ***P<0.001, **** P<0.0001;

[0039] Figure 11 This is a comparative graph showing the effects of different doses of the traditional Chinese medicine composition on apoptosis in plaque cells compared to the high-fat diet group in Example 1. * P<0.05, ** P<0.01, *** P<0.001;

[0040] Figure 12 This is a comparative graph showing the effects of different doses of the traditional Chinese medicine composition on vascular endothelial function in Example 1, compared to the high-fat diet group. * P<0.05, *** P<0.001;

[0041] Figure 13 This is a comparison chart of different doses of the traditional Chinese medicine composition inhibiting the inflammatory response in plaques in Example 1;

[0042] In each figure, C represents the control group (HFD); S represents the atorvastatin treatment group at the usual dose; A1 represents the treatment group at the usual dose of the traditional Chinese medicine composition in Example 1; and A2 represents the treatment group at the high dose of the traditional Chinese medicine composition in Example 1. Detailed Implementation

[0043] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and 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.

[0044] Example 1

[0045] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 110g Gynostemma pentaphyllum, 250g Ganoderma lucidum, 260g Paeonia lactiflora, 180g Citrus aurantium and 200g Allium macrostemon (11 parts Gynostemma pentaphyllum, 25 parts Ganoderma lucidum, 26 parts Paeonia lactiflora, 18 parts Citrus aurantium and 20 parts Allium macrostemon).

[0046] Preparation method of traditional Chinese medicine composition:

[0047] (1) Weigh out Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon and grind them into coarse powder. Add 15 times the amount of water for the first extraction for 1.5 hours, add 12 times the amount of water for the second extraction for 1 hour, combine the extracts to obtain extract 1, filter, concentrate the filtrate under reduced pressure at 60℃ to a relative density of 1.05, dry, and obtain dried product 1.

[0048] (2) Weigh out Ganoderma lucidum and Paeonia lactiflora and crush them into coarse powder. First, add 16 times the amount of 45% ethanol solution and extract for 2 hours. Second, add 12 times the amount of 70% ethanol solution and extract for 1.5 hours. Combine the extracts to obtain extract 2. Filter and concentrate the filtrate under reduced pressure at 60℃ to a relative density of 1.05. Dry to obtain dried product 2.

[0049] (3) Mix the dried material 1 and dried material 2 to make a powder.

[0050] Example 2

[0051] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 120g Gynostemma pentaphyllum, 250g Ganoderma lucidum, 280g Paeonia lactiflora, 150g Citrus aurantium and 220g Allium macrostemon (12 parts Gynostemma pentaphyllum, 25 parts Ganoderma lucidum, 28 parts Paeonia lactiflora, 15 parts Citrus aurantium and 22 parts Allium macrostemon).

[0052] Preparation method of traditional Chinese medicine composition:

[0053] (1) Weigh out Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon and grind them into coarse powder. Add 16 times the amount of water for the first extraction for 1.7 h, add 13 times the amount of water for the second extraction for 0.8 h, combine the extracts to obtain extract 1, filter, concentrate the filtrate under reduced pressure at 65℃ to a relative density of 1.15, dry to obtain dried product 1;

[0054] (2) Weigh out Ganoderma lucidum and Paeonia lactiflora and crush them into coarse powder. First, extract with 17 times the amount of 50% ethanol solution for 2.2 hours. Second, extract with 13 times the amount of 75% ethanol solution for 1.7 hours. Combine the extracts to obtain extract 2. Filter the extract and concentrate it under reduced pressure at 65°C to a relative density of 1.15. Dry the extract to obtain dried product 2.

[0055] (3) Mix the dried material 1 and dried material 2 to make a powder.

[0056] Example 3

[0057] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 310g Gynostemma pentaphyllum, 230g Ganoderma lucidum, 260g Paeonia lactiflora, 60g Citrus aurantium and 140g Allium macrostemon (31 parts Gynostemma pentaphyllum, 23 parts Ganoderma lucidum, 26 parts Paeonia lactiflora, 6 parts Citrus aurantium and 14 parts Allium macrostemon).

[0058] Preparation method of traditional Chinese medicine composition:

[0059] (1) Weigh out Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon and grind them into coarse powder. Add 14 times the amount of water for the first extraction for 1.3 hours, add 11 times the amount of water for the second extraction for 1.2 hours, combine the extracts to obtain extract 1, filter, concentrate the filtrate under reduced pressure at 55℃ to a relative density of 1.05, dry, and obtain dried product 1.

[0060] (2) Weigh out Ganoderma lucidum and Paeonia lactiflora and crush them into coarse powder. First, add 15 times the amount of 40% ethanol solution and extract for 1.8h. Second, add 11 times the amount of 65% ethanol solution and extract for 1.3h. Combine the extracts to obtain extract 2. Filter and concentrate the filtrate under reduced pressure at 55℃ to a relative density of 1.01. Dry to obtain dried product 2.

[0061] (3) Mix the dried material 1 and dried material 2 to make a powder.

[0062] Example 4

[0063] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 210g Gynostemma pentaphyllum, 130g Ganoderma lucidum, 120g Paeonia lactiflora, 280g Citrus aurantium and 260g Allium macrostemon (21 parts Gynostemma pentaphyllum, 13 parts Ganoderma lucidum, 12 parts Paeonia lactiflora, 28 parts Citrus aurantium and 26 parts Allium macrostemon).

[0064] The preparation method of the traditional Chinese medicine composition is the same as that in Example 1.

[0065] Example 5

[0066] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 50g Gynostemma pentaphyllum, 470g Ganoderma lucidum, 120g Paeonia lactiflora, 160g Citrus aurantium and 200g Allium macrostemon (5 parts Gynostemma pentaphyllum, 47 parts Ganoderma lucidum, 12 parts Paeonia lactiflora, 16 parts Citrus aurantium and 20 parts Allium macrostemon).

[0067] The preparation method of the traditional Chinese medicine composition is the same as that in Example 1.

[0068] Comparative Example 1

[0069] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 310g Gynostemma pentaphyllum, 250g Ganoderma lucidum, 260g Paeonia lactiflora and 180g Citrus aurantium (31 parts Gynostemma pentaphyllum, 25 parts Ganoderma lucidum, 26 parts Paeonia lactiflora and 18 parts Citrus aurantium).

[0070] Preparation method of traditional Chinese medicine composition:

[0071] (1) Weigh out Gynostemma pentaphyllum and Citrus aurantium and crush them into coarse powder. Add 15 times the amount of water for the first extraction for 1.5 hours, and add 12 times the amount of water for the second extraction for 1 hour. Combine the extracts to obtain extract 1. Filter the extract and concentrate it under reduced pressure at 60°C to a relative density of 1.05. Dry the extract to obtain dried product 1.

[0072] (2) Weigh out Ganoderma lucidum and Paeonia lactiflora and crush them into coarse powder. First, add 16 times the amount of 45% ethanol solution and extract for 2 hours. Second, add 12 times the amount of 70% ethanol solution and extract for 1.5 hours. Combine the extracts to obtain extract 2. Filter and concentrate the filtrate under reduced pressure at 60℃ to a relative density of 1.05. Dry to obtain dried product 2.

[0073] (3) Mix the dried material 1 and dried material 2 to make a powder.

[0074] Comparative Example 2

[0075] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 250g Ganoderma lucidum, 260g Paeonia lactiflora, 180g Citrus aurantium and 310g Allium macrostemon (25 parts Ganoderma lucidum, 26 parts Paeonia lactiflora, 18 parts Citrus aurantium and 31 parts Allium macrostemon).

[0076] Preparation method of traditional Chinese medicine composition:

[0077] (1) Weigh out the bitter orange peel and the white allium macrostemon and crush them into coarse powder. Add 15 times the amount of water for the first extraction for 1.5 hours, and add 12 times the amount of water for the second extraction for 1 hour. Combine the extracts to obtain extract 1. Filter the extract and concentrate it under reduced pressure at 60°C to a relative density of 1.05. Dry the extract to obtain dried product 1.

[0078] (2) Weigh out Ganoderma lucidum and Paeonia lactiflora and crush them into coarse powder. First, add 16 times the amount of 45% ethanol solution and extract for 2 hours. Second, add 12 times the amount of 70% ethanol solution and extract for 1.5 hours. Combine the extracts to obtain extract 2. Filter and concentrate the filtrate under reduced pressure at 60℃ to a relative density of 1.05. Dry to obtain dried product 2.

[0079] (3) Mix the dried material 1 and dried material 2 to make a powder.

[0080] Comparative Example 3

[0081] A traditional Chinese medicine composition for improving atherosclerosis, the raw materials are: 110g Gynostemma pentaphyllum, 250g Ganoderma lucidum, 260g Paeonia lactiflora, 180g Citrus aurantium and 300g Allium macrostemon (11 parts Gynostemma pentaphyllum, 25 parts Ganoderma lucidum, 26 parts Paeonia lactiflora, 18 parts Citrus aurantium and 30 parts Allium macrostemon).

[0082] The preparation method of the traditional Chinese medicine composition is the same as that in Example 1.

[0083] Comparative Example 4

[0084] A traditional Chinese medicine composition for improving atherosclerosis, using the same raw materials as in Example 1.

[0085] Preparation method of traditional Chinese medicine composition:

[0086] Weigh out Gynostemma pentaphyllum, Citrus aurantium, Allium macrostemon powder, Ganoderma lucidum, and Paeonia lactiflora and crush them into coarse powder. First, extract with 16 times the amount of 45% ethanol solution for 2 hours. Second, extract with 12 times the amount of 70% ethanol solution for 1.5 hours. Combine the extracts, filter, concentrate the filtrate under reduced pressure at 60℃ to a relative density of 1.05, dry, and prepare a powder.

[0087] The Chinese medicine compositions and ratios of Examples 1-9 and Comparative Examples 1-4 are shown in Table 1.

[0088] Table 1 Composition and weight ratio of Chinese medicine compositions in Examples 1-9 and Comparative Examples 1-4

[0089] Example 1 11 25 26 18 20 Example 2 12 25 28 15 22 Example 3 31 23 26 6 4 Example 4 21 13 12 28 26 Example 5 5 47 12 16 20 Comparative Example 1 31 25 26 18 - Comparative Example 2 - 25 26 18 31 Comparative Example 3 11 25 26 40 20 Comparative Example 4 11 25 26 18 20

[0090] Effect test

[0091] Test Example 1 Evaluation of the efficacy of improving atherosclerosis

[0092] 1.1 Experimental animals

[0093] 8-week-old male ApoE- / - mice, experimental animal license number: SCXK(Shanghai)2022-0004. They were housed in a clean-grade animal room at a temperature of 18-26 °C, a relative humidity of 50-60%, with 12 hours of alternating light and dark and ventilation 22 times per hour.

[0094] 1.2 Experimental drugs

[0095] Atorvastatin, purchased from Pfizer Pharmaceuticals Limited; Chinese medicine compositions of Examples 1-5 and Comparative Examples 1-4.

[0096] 2. Grouping and administration

[0097] 8-week-old male ApoE- / - mice were selected and randomly divided into 15 groups, with 15 mice in each group (n = ), 5 mice in each cage. Control group 1 was the high-fat diet group, fed a high-fat diet (HFD, 21% fat, 0.5% cholesterol, MD 15HL, medicience Ltd.) for 19 weeks; Control group 2 was the atorvastatin conventional-dose treatment group (calculated according to the adult dose of 20 mg, the mouse administration dose was 2.6 mg / kg / d), atorvastatin was administered by gavage + fed HFD for 19 weeks; The experimental groups were the Chinese medicine compositions of Examples 1-5 and Comparative Examples 1-4, the liquid medicine was administered by gavage + fed HFD for 19 weeks (calculated according to the adult dose of 20 mg, the mouse administration dose was 1227.2 mg / kg / d). The conversion formula for the equivalent dose of 70 kg adults and mice: mouse dose = adult dose × 9.1 mg / kg was used to calculate the mouse gavage dose. The clinical conventional dose of atorvastatin is 0.5 mg / kg·d, and the clinical conventional dose of the Chinese medicine composition is 236 mg / kg·d.

[0098] 3. Experimental methods

[0099] Eight-week-old male ApoE- / - mice were grouped, modeled, and administered drugs as described above. After 19 weeks of rearing, modeling was completed, and the entire aorta of each group was dissected. Oil Red staining, HE staining, and a calcium detection kit (S1063S, Shanghai Beyotime Biotechnology Co., Ltd.) were used to assess the plaque area, plaque stability, and degree of plaque calcification in the entire aorta of each group of mice. The efficacy of the traditional Chinese medicine compositions of Examples 1-5 and Comparative Examples 1-4 on atherosclerosis was evaluated.

[0100] 3.1 Effect on total aortic plaque area

[0101] After mouse modeling was completed, the mice were sacrificed, and the entire aorta was dissected. The dissected aorta was rinsed in physiological saline, then fixed in 4% paraformaldehyde at room temperature for 10 min, and residual fat adhering to the aortic lumen was removed. The aorta was then immersed in Oil Red O working solution for Oil Red staining. The plaque area of ​​the entire aorta was quantitatively analyzed using ImageJ software, and the percentage (%) of the total aortic plaque area was obtained by dividing by the corresponding aortic wall area.

[0102] 3.2 Effect on plaque stability

[0103] After mouse modeling was completed, the mice were sacrificed, the entire aorta was dissected, and frozen sections of the aortic root were stained with hematoxylin for 2 minutes and then gently rinsed with running water. The frozen sections were then stained with eosin for 30 seconds. The necrotic core area in the plaque was counted and the area of ​​the necrotic core was analyzed to assess the stability of the plaque.

[0104] 3.3 Effect on the degree of plaque calcification

[0105] After mouse modeling was completed, mice were euthanized, and the entire aorta was dissected. Frozen sections of the aortic root were stained with Alizarin Red S for 1 hour, differentiated with McGee-Russell differentiation solution for a few seconds, and then stained with hematoxylin for 10 minutes. Calcium deposition in the plaques was counted to assess plaque stability. The amount of calcium deposition in the aorta was detected using a calcium assay kit (CalciumColorimetric Assay Ki, Sigma-Aldrich Inc.), and the calcium content per unit mass of aorta was calculated.

[0106] The test results are shown in Table 2.

[0107] Table 2. Effects of Traditional Chinese Medicine Compositions on Aortic Atherosclerosis in Mice

[0108]

[0109] Test Example 2: Determination of Active Ingredients and Calculation of Active Ingredient Transfer Rate

[0110] Gynostemma pentaphyllum saponins XVII, paeoniflorin, ganoderic acid A, and naringin are the main active ingredients in the traditional Chinese medicine composition. Their contents were determined using the following methods.

[0111] 1. Determination of Gynostemma pentaphyllum saponin XVII content

[0112] Chromatographic conditions: An Agilent ZORBAX SB-C18 (250 mm × 4.6 mm, 5 μm) column was used; the mobile phase was 0.3% formic acid solution-acetonitrile (30:70); the flow rate was 1.5 ml / min; the detection wavelength was 203 nm; the column temperature was 35 ℃; and the injection volume was 10 μl.

[0113] Preparation of reference solution: Accurately weigh an appropriate amount of Gynostemma pentaphyllum saponin XVII reference standard and add methanol to prepare a reference stock solution with a concentration of 3200.01 μg / ml.

[0114] Preparation of the test solution: Accurately weigh 0.2 g of sample powder (passed through a No. 2 sieve) dried to constant weight at 60℃, place it in a stoppered conical flask, accurately add 25 ml of methanol, sonicate (100 W, 100 Hz) for 30 min, cool, evaporate the filtrate to dryness, dissolve in water, extract with water-saturated n-butanol in small amounts several times until colorless, combine the extracts, wash with ammonia test solution, evaporate to dryness under reduced pressure, add methanol to make up to 100 ml, shake well, filter through a 0.22 μm microporous membrane to obtain the test solution.

[0115] Determination method: Accurately pipette 5 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0116] 2. Determination of paeoniflorin content

[0117] Chromatographic conditions: An Agilent ZORBAX SB-C18 (250 mm × 4.6 mm, 5 μm) column was used; the mobile phase was methanol-0.05 mol / L potassium dihydrogen phosphate solution (40:65); the detection wavelength was 230 nm. The theoretical plate number, calculated based on the paeoniflorin peak, should be no less than 3000.

[0118] Preparation of reference solution: Take an appropriate amount of paeoniflorin reference standard that has been dried in a phosphorus pentoxide vacuum desiccator for 36 hours, accurately weigh it, and add methanol to prepare a solution containing 0.5 mg per ml.

[0119] Preparation of the test solution: Take about 0.5g of crude powder of this product, accurately weigh it, place it in a stoppered conical flask, accurately add 25ml of methanol, weigh it, soak for 4h, sonicate for 20min, cool, weigh it again, make up the weight loss with methanol, shake well, filter, and take the filtrate to obtain the test solution.

[0120] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0121] 3. Determination of Ganoderic A Content

[0122] Chromatographic conditions: The column was a CORTECS T3 column (4.6 mm × 150 mm, 2.7 μm); acetonitrile was used as mobile phase A, and 0.1% formic acid solution was used as mobile phase B, with gradient elution performed according to the specifications in Table 2; the flow rate was 1.0 mL / min; the column temperature was 25℃; and the detection wavelength was 254 nm. The theoretical plate number, calculated based on the Ganoderma lucidum acid A peak, should be no less than 5000.

[0123] Preparation of reference solution: Take an appropriate amount of ganoderic acid A reference standard, accurately weigh it, and add methanol to prepare a solution containing 30 μg per ml.

[0124] Preparation of the test solution: Accurately weigh 2g of sample powder and place it in an Erlenmeyer flask. Add 40mL of methanol and extract by sonication for 30min. Cool and filter into a 50mL volumetric flask. Dilute to the mark with methanol and mix well. Filter through a microporous membrane (0.45μm) and collect the filtrate.

[0125] Assay: Accurately pipette 5 μl of the reference solution and the test solution into the liquid chromatograph and determine.

[0126] 4. Determination of naringin content

[0127] Chromatographic conditions: An Agilent ZORBAX SB-C18 (250 mm × 4.6 mm, 5 μm) column was used; acetonitrile-water (20:80) (pH adjusted to 3 with phosphoric acid) was used as the mobile phase; the detection wavelength was 283 nm. The theoretical plate number, calculated based on the naringin peak, should be no less than 3000.

[0128] Preparation of reference solutions: Take an appropriate amount of naringin reference standard, accurately weigh it, and add methanol to prepare solutions containing 80 μg each of naringin and neohesperidin per ml.

[0129] Preparation of the test solution: Accurately weigh approximately 0.2 g of the crude powder and place it in a stoppered conical flask. Accurately add 50 ml of methanol, weigh the solution, heat under reflux for 1.5 h, cool, weigh again, and replenish the lost weight with methanol. Shake well and filter. Accurately measure 10 ml of the filtrate, place it in a 25 ml volumetric flask, add methanol to the mark, and shake well.

[0130] Assay: Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph and determine the content. The active ingredient content and effective ingredient transfer rate of the traditional Chinese medicine composition in Example 1 are shown in Table 3.

[0131] Table 3. Content of active ingredients and transfer rate of effective ingredients

[0132] Gynostemma pentaphyllum saponins XVII 1.01 82.3 paeoniflorin 1.84 89.6 Ganoderic acid A 0.05 79.6 Naringin 4.05 74.7

[0133] Test Example 3: Evaluation of the efficacy of drugs in improving hyperlipidemia and atherosclerosis

[0134] 1. Animal grouping and drug administration

[0135] Eight-week-old male ApoE - / - Mice were randomly divided into 4 groups of 15 mice each (n=15), with 5 mice per cage. Example 1 (normal and high doses) and atorvastatin (ATO) were calculated using the conversion formula for equivalent doses in mice to 70kg adults: mouse dose = adult dose × 9.1mg / kg. The commonly used clinical dose of atorvastatin is 0.5mg / kg·d, and the commonly used clinical dose of the traditional Chinese medicine composition is 236mg / kg·d. This study verified whether the traditional Chinese medicine composition of the present invention could improve hyperlipidemia and AS.

[0136] Group 1 (Control, C): The control group (HFD) was fed a high-fat diet (HFD, 21% fat, 0.5% cholesterol, MD12015HL, medicine Ltd.) for 19 weeks.

[0137] Group 2 (Statin, S): Atorvastatin treatment group at the usual dose (calculated based on the adult dose of 20 mg, the mouse dose was 2.6 mg / kg d), atorvastatin by gavage + HFD for 19 weeks;

[0138] Group 3 (normal dose of Example 1, A1): The treatment group (A1) of the commonly used dose of the Chinese medicine composition in Example 1 was administered the drug solution by gavage (calculated based on the adult dose of 20mg, the dose administered to mice was 1227.2mg / kg d) + fed with HFD for 19 weeks;

[0139] Group 4 (High dose of Example 1, A2): High dose treatment group of the traditional Chinese medicine composition in Example 1 (A2) (calculated by A1 dose × 2) The drug solution was administered by gavage (calculated based on the adult dose of 20mg, the mouse administration dose was 2454.4mg / kg d) + fed HFD for 19 weeks.

[0140] 2. Experimental Methods

[0141] 2.1 Effects on body weight in mice fed a high-fat diet

[0142] After the mouse model was completed, the weight of each group of mice was weighed and recorded. The weight change curve was observed, and the weight during the experiment and the weight at the end of the experiment were analyzed to determine whether there were any differences and whether the herbal composition in Example 1 could improve the obesity symptoms induced by high fat.

[0143] 2.2 Effects on hepatic lipid metabolism

[0144] After the mouse model was completed, the mice were sacrificed, and the livers of each group of mice were taken. The frozen sections of the mouse liver were stained with HE to observe whether the mouse liver tissue was intact and clear, and to observe the inflammation index, hepatocyte ballooning degeneration and necrosis index of the mouse liver. The mouse liver was stained with Oil Red to observe whether there was lipid accumulation in the mouse liver tissue and to evaluate the trend of lipid metabolism disorder after liver function injury.

[0145] 2.3 Effects on blood lipids

[0146] After mouse modeling was completed, whole blood was collected from each group of mice and centrifuged to obtain serum. The levels of TG and LDL were detected using a mouse triglyceride (TG) ELISA kit (COBIO Biotech, CB10296-Mu) and a mouse low-density lipoprotein (LDL) ELISA kit (COBIO Biotech, CB10259-Mu).

[0147] 2.4 Effects on oxidative stress

[0148] After mouse modeling was completed, the mice were sacrificed, and whole blood was collected from each group of mice and centrifuged to obtain serum. The superoxide dismutase (SOD) level in the serum of mice after drug administration was detected using a mouse superoxide dismutase (SOD) ELISA kit (COAB Biotech, CB10221-Mu).

[0149] 2.5 Effects on the area and number of plaques in the entire aorta

[0150] After mouse modeling was completed, the mice were sacrificed, and the dissected aorta was rinsed in physiological saline, then fixed in 4% paraformaldehyde at room temperature for 10 min, and residual fat adhering to the aortic lumen was removed. The aorta was then immersed in Oil Red O working solution for Oil Red staining, and the plaque area of ​​the entire aorta was quantitatively analyzed using ImageJ software. The plaque area percentage of the entire aorta was obtained by dividing the area by the corresponding aortic wall area.

[0151] 2.6 Effect on the area of ​​plaque in the entire aortic root

[0152] After the mouse model was completed, the mice were sacrificed, frozen sections of the aortic root were prepared, stained with Oil Red dye, the plaque area was counted, the degree of valve stenosis was analyzed, and it was determined whether the plaque at the arterial valve had an inhibitory effect.

[0153] 2.7 Effect on plaque stability

[0154] After mouse modeling was completed, the mice were sacrificed, the aorta was dissected, and frozen sections of the aortic root were stained with hematoxylin for 2 minutes and then gently rinsed with running water. The frozen sections were then stained with eosin for 30 seconds. The necrotic core area in the plaque was counted, and the area of ​​the necrotic core and the thickness of the fibrous cap were analyzed to assess the stability of the plaque.

[0155] 2.8 Effect on foam cell content in plaques

[0156] After mouse modeling was completed, mice were euthanized, and their aortas were dissected. Aortic root sections were subjected to immunofluorescence co-staining with cell proliferation marker Ki67 and macrophage marker CD68. Frozen sections of the aortic root were blocked with 2% BSA for 1 hour, then diluted primary antibody was added, and the sections were incubated overnight at 4°C. The next day, the frozen sections were washed three times with PBS for 10 minutes each time, and then incubated with secondary antibody at 37°C for 1 hour. The sections were then washed three more times with PBS and sealed in the dark. Subsequently, images were taken using a fluorescence microscope, and antibody expression was statistically analyzed. The effect of the herbal composition in Example 1 on the foam cell content in plaques was indirectly analyzed.

[0157] 2.9 Effect on the content of smooth muscle cells in plaques

[0158] After mouse modeling was completed, mice were euthanized, and their aortas were dissected. Aortic root sections were subjected to immunofluorescence co-staining with the proliferation marker Ki67 and the VSMC marker αSMA. Frozen sections of the aortic root were blocked with 2% BSA for 1 hour, then diluted primary antibody was added, and the sections were incubated overnight at 4°C. The next day, the frozen sections were washed three times with PBS for 10 minutes each time, and then incubated with secondary antibody at 37°C for 1 hour. The sections were then washed three more times with PBS and sealed in the dark. Subsequently, images were taken using a fluorescence microscope, and antibody expression was statistically analyzed. The effect of the herbal composition in Example 1 on the smooth muscle cell content in plaques was indirectly analyzed.

[0159] 2.10 Effect on plaque calcification degree

[0160] After mouse modeling was completed, mice were euthanized, and their aortas were dissected. Frozen sections of the aortic root were stained with Alizarin Red S for 1 hour, differentiated for a few seconds with McGee-Russell differentiation solution, and then stained with hematoxylin for 10 minutes. Calcium deposition in the plaques was counted to assess plaque stability. The amount of calcium deposition in the aorta was detected using a calcium assay kit (CalciumColorimetric Assay Ki, Sigma-Aldrich Inc.), and the calcium content per unit mass of aorta was calculated.

[0161] 2.11 Effects on apoptosis within plaque cells

[0162] After mouse modeling was completed, mice were euthanized, and their aortas were dissected. Proteinasek solution was diluted 1:100 with PBS to a final concentration of 2 mg / ml (2 mg / ml), resulting in a final concentration of 20 μg / ml. After incubation at room temperature for 10 min, the mice were washed with PBS, and 5x Equilibrium Buffer was diluted 5-fold with ddH2O and incubated at room temperature for 20 min. During incubation, TdT buffer was prepared, the liquid was discarded, the mice were placed in a humidified chamber, 50 ml of TdT buffer was added, and the chamber was incubated at 37°C for 1 h. Mounting was performed using DAPI-containing anti-fluorescence quenching mounting media, and the images were obtained under an upright fluorescence microscope. The proportion of apoptotic cells in the plaques was calculated.

[0163] 2.12 Effects on vascular endothelial function

[0164] After mouse modeling was completed, the mice were euthanized and their aortas were dissected. Immunofluorescence staining was performed using the same method as in 2.9. Endothelium-dependent vasodilatory indices and independent vasodilatory-related indices (iNOS) were detected by immunofluorescence co-localization staining (co-stained with CD31) to evaluate whether vasodilatory function was improved. Immunofluorescence co-localization was also used to detect the expression of endothelial cell adhesion factor ICAM-1, confirming that the herbal composition in Example 1 could inhibit endothelial cell activation.

[0165] 2.13 Inhibits inflammatory response in plaques

[0166] After mouse modeling was completed, the mice were sacrificed, the aorta was dissected, and immunofluorescence staining was performed using the same method as in 2.9. Further immunofluorescence co-staining was performed on the macrophage marker CD68 and the inflammatory factor IL-1 in the aortic root to detect the expression of inflammatory factors in foam cells in the plaque.

[0167] 3. Experimental Results

[0168] The improvement in blood lipids in each group of mice is shown in the figure. Figure 1-4 .

[0169] Weight: such as Figure 1 As shown, compared with the high-fat diet group, the body weight of mice in all groups decreased, with significant differences in body weight changes between atorvastatin (S group) and the high-dose group of Example 1 (A2).

[0170] Oil red staining results showed that a large amount of red lipid deposition was observed in the high-fat diet group. Compared with the high-fat diet group, a small amount of red lipid was observed in the S group and the normal dose group (A1) of Example 1. HE staining results showed that under high-fat feeding conditions, the high-fat diet group had a large accumulation of lipids in the liver and severe gas bubble-like lesions. At the same time, hepatocytes were hypertrophied and their arrangement was significantly disordered. After drug intervention, compared with the high-fat diet group, the liver lipid levels, hepatocyte morphology, and gas bubble-like lesions in the S group and the normal dose group (A1) of Example 1 were significantly improved. The S group and the normal dose group (A1) of Example 1 were able to improve liver lipid accumulation and liver pathological changes caused by high fat intake. Figure 2 (A and B in the text).

[0171] Whole blood was collected and centrifuged to obtain serum. TG and LDL levels were then measured. The results showed that... Figure 3 As shown, in terms of TG regulation, the normal dose group (A1) in Example 1 showed a trend of decreasing TG, but the difference was not statistically significant. The high dose group (A2) in Example 1 significantly reduced TG levels. Figure 3 In terms of LDL regulation, both the normal dose group (A1) and the high dose group (A2) in Example 1 effectively reduced LDL levels. Figure 3 (B in the middle).

[0172] The study investigated the levels of key enzymes related to oxidative stress in mouse serum after drug administration. Results showed that different doses of the herbal composition from Example 1 resulted in varying degrees of increase in SOD levels, indicating a significant therapeutic effect. Figure 4 ).

[0173] The aortic atherosclerosis status of mice in each group is shown in the figure. Figure 5-13 Compared with the high-fat diet group, the total aortic plaque area in mice treated with different doses in Example 1 was significantly reduced. Figure 5 The ratio of necrotic core area to plaque area and the degree of aortic plaque calcification were significantly lower in the aortic root plaque group than in the high-fat diet group, while the ratio of fibrous cap to plaque area was significantly higher in the aortic root plaque group. Figure 6 , Figure 7 Compared with the high-fat diet group, the number of Ki67 / CD68 double-positive macrophages in the plaques of the S group and the normal dose group of Example 1 (A1) was significantly reduced. Figure 8 The herbal composition of this invention and atorvastatin can both reduce the number of macrophages in plaques and indirectly inhibit the formation of foam cells.

[0174] Compared with the high-fat diet group, αSMA expression in plaques was significantly increased in the atorvastatin and normal dose groups (A1) of Example 1 under drug intervention. Figure 9 Atorvastatin (Group S) and the herbal composition of Example 1 significantly increased the VSMC content in plaques and significantly decreased the calcium content in the aorta. Figure 10Meanwhile, compared with the high-fat diet group, the apoptotic area within the plaque was reduced in both the atorvastatin (S group) and the normal dose group of Example 1 (A1). Figure 11 All of these can reduce cell apoptosis in AS plaques; they can significantly reduce the expression of vascular endothelial adhesion factors and inhibit endothelial cell activation. Figure 12 Furthermore, both Group S and the normal dose group of Example 1 (A1) inhibited the inflammatory response in the plaque. Figure 13 It inhibits the inflammatory microenvironment in plaques.

[0175] Experimental results showed that the normal dose group (A1) in Example 1 had a significant effect on improving blood lipids and aortic atherosclerosis, and this effect was dose-dependent.

[0176] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the patent scope of the present invention. All equivalent implementations or modifications that do not depart from the present invention should be included within the scope of the technical solution of the present invention.

Claims

1. A traditional Chinese medicine composition, characterized in that, The raw materials of the traditional Chinese medicine composition, by weight, are: 1-31 parts of Gynostemma pentaphyllum, 6-50 parts of Ganoderma lucidum, 12-40 parts of Paeonia lactiflora, 6-34 parts of Citrus aurantium, and 4-28 parts of Allium macrostemon; the specific preparation method of the composition is as follows: (1) Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon powder were extracted with water to obtain extract 1, which was filtered, concentrated and dried to obtain dried product 1; (2) Extract Ganoderma lucidum and Paeonia lactiflora with alcohol to obtain extract 2, filter, concentrate and dry to obtain dried product 2; (3) Mix dried substance 1 and dried substance 2 to obtain the product; The water extraction step in step (1) is as follows: add 10-20 times the volume of water to the powders of Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon, extract 1-3 times, each extraction for 1-3 hours, and combine to obtain extract 1; the concentration temperature is 50-70 ℃, and the concentration is carried out to a relative density of 1.01-1.25; The alcohol extraction step in step (2) is as follows: add 12-18 times the volume of ethanol solution with a mass concentration of 30%-80% to the powder of Ganoderma lucidum and Paeonia lactiflora, extract 1-3 times, each extraction for 1-3 hours, and combine to obtain extract 2; the concentration temperature is 50-70℃, and the concentration is carried out to a relative density of 1.01-1.

25.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The raw materials of the traditional Chinese medicine composition, by weight, are: 5-31 parts of Gynostemma pentaphyllum, 13-47 parts of Ganoderma lucidum, 12-30 parts of Paeonia lactiflora, 6-28 parts of Citrus aurantium, and 14-26 parts of Allium macrostemon.

3. The traditional Chinese medicine composition according to claim 1, characterized in that, The raw materials of the traditional Chinese medicine composition, by weight, are: 10-12 parts of Gynostemma pentaphyllum, 22-28 parts of Ganoderma lucidum, 25-28 parts of Paeonia lactiflora, 15-22 parts of Citrus aurantium and 18-22 parts of Allium macrostemon.

4. A method for preparing the traditional Chinese medicine composition according to any one of claims 1-3, characterized in that, The specific steps are as follows: (1) Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon powder were extracted with water to obtain extract 1, which was filtered, concentrated and dried to obtain dried product 1; (2) Extract Ganoderma lucidum and Paeonia lactiflora with alcohol to obtain extract 2, filter, concentrate and dry to obtain dried product 2; (3) Mix dried substance 1 and dried substance 2 to obtain the product; The water extraction step in step (1) is as follows: add 10-20 times the volume of water to the powders of Gynostemma pentaphyllum, Citrus aurantium and Allium macrostemon, extract 1-3 times, each extraction for 1-3 hours, and combine to obtain extract 1; the concentration temperature is 50-70 ℃, and the concentration is carried out to a relative density of 1.01-1.25; The alcohol extraction step in step (2) is as follows: add 12-18 times the volume of ethanol solution with a mass concentration of 30%-80% to the powder of Ganoderma lucidum and Paeonia lactiflora, extract 1-3 times, each extraction for 1-3 hours, and combine to obtain extract 2; the concentration temperature is 50-70℃, and the concentration is carried out to a relative density of 1.01-1.

25.

5. The preparation method according to claim 4, characterized in that, In step (1), the water extraction is performed twice. The first extraction is performed by adding 14-16 times the volume of water and extracting for 1.3-1.7 h. The second extraction is performed by adding 11-13 times the volume of water and extracting for 0.8-1.2 h. The concentration temperature is 55-65 ℃, and the concentration is performed until the relative density is 1.01-1.

15. The alcohol extraction in step (2) is performed twice. The first extraction is performed by adding 15-17 times the volume of ethanol solution with a mass concentration of 40%-50% for 1.8-2.2 h. The second extraction is performed by adding 11-13 times the volume of ethanol solution with a mass concentration of 65%-75% for 1.3-1.7 h. The concentration temperature is 55-65 ℃, and the product is concentrated to a relative density of 1.01-1.

15.

6. The use of the composition according to any one of claims 1-3 in the preparation of a medicament for the prevention and / or treatment of hyperlipidemia and its complications.

7. The application according to claim 6, characterized in that, The dosage form of the drug is selected from any one of tablets, granules, powders and capsules; the drug is used to lower blood lipids and improve lipid metabolism disorders, or the drug is used to prevent and / or treat atherosclerosis.

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