Traditional Chinese medicine composition for preventing and treating atherosclerosis as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for preventing and treating atherosclerosis and a preparation method and application thereof.The traditional Chinese medicine composition is prepared from, by weight, 6-30 parts of ganoderma lucidum, 5-15 parts of curcuma zedoary, 6-30 parts of angelica sinensis, 5-20 parts of ligusticum wallichii and 9-30 parts of polygonum cuspidatum. The preparation method comprises the following steps: soaking the lucid ganoderma, the curcuma zedoary, the angelica sinensis, the ligusticum wallichii and the polygonum cuspidatum in water, and decocting; and uniformly mixing the decoction obtained by decocting, or concentrating to obtain thick paste, or concentrating and drying to obtain the traditional Chinese medicine composition. The key pathogenesis of atherosclerosis is explored, the prescription is accurately pushed, the anti-atherosclerosis traditional Chinese medicine prescription which is rigorous in compatibility and synergistic in effect is achieved, and the traditional Chinese medicine composition has the remarkable effect of treating and / or preventing atherosclerosis.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for preventing and treating atherosclerosis, a preparation method and an application thereof. Background Art

[0002] Atherosclerosis (AS) is the primary pathological basis of ischemic cardio-cerebrovascular diseases. Although modern medicine has made significant progress in anti-AS by regulating lipid metabolism and antiplatelet aggregation, the incidence of ischemic cardiovascular events has not been effectively controlled. The progression of AS plaques is the key. How to further inhibit or even reduce the occurrence and development of AS plaques remains a major need in the prevention and treatment of cardio-cerebrovascular diseases.

[0003] So far, the pathogenesis of AS has not been fully clarified, including lipid infiltration theory, vascular smooth muscle cell cloning theory, oxidative stress theory, platelet hyperfunction theory, and inflammation theory, etc. Since the direct pathological manifestation of AS is an abnormal response of the blood vessel wall to various injuries, with characteristics such as inflammatory degeneration, exudation, and hyperplasia, and the advantages of traditional Chinese medicine in anti-inflammation, immune regulation, softening and shrinking plaques, and protecting blood vessels are becoming increasingly obvious.

[0004] AS belongs to the categories of diseases such as "vertigo", "chest impediment", and "pulse impediment" in traditional Chinese medicine. Ancient traditional Chinese medicine classics, such as "Huangdi Neijing", "Synopsis of the Golden Chamber", and "Treatise on the Causes and Symptoms of Various Diseases", have rich discussions on the related symptoms of this disease. And the pathological changes of the occurrence and development process of AS, such as (abnormal lipid metabolism + endothelial injury) - inflammatory reaction - lipid deposition - fibrous plaque - lumen stenosis, are similar to "blood vessel mass" in traditional Chinese medicine. Therefore, blood vessel deficiency - mass stasis obstruction is the key pathogenesis of the occurrence and development of AS.

[0005] Chinese Patent Application CN 116966252A discloses a traditional Chinese medicine composition for treating menopausal atherosclerosis. The key points of its technical solution are: the composition is composed of 10 - 20 g of Dioscorea nipponica, 5 - 15 g of Curcuma zedoaria, 5 - 15 g of Paeonia lactiflora, and 3 - 7 g of Coptis chinensis. Since menopause is a special physiological period in the female population, the treatment and prevention of general AS are not clear, and the medication is mainly "attacking", and the effect on the pathological state of blood vessels is not clear, and its treatment effect still has limitations.

[0006] Therefore, for the current traditional Chinese medicine treatment of AS, there is still a need for a traditional Chinese medicine composition that can improve the disease from multiple angles and specifically, so as to achieve a more significant and efficient treatment effect. Summary of the Invention

[0007] In view of the deficiencies in the prior art, the present invention provides a Chinese medicine composition for preventing and treating atherosclerosis, a preparation method and application thereof. By exploring the key pathogenesis of AS and accurately considering the prescription, a rigorous and synergistic anti-AS Chinese medicine prescription is achieved, which has a significant therapeutic and / or preventive effect on AS.

[0008] In order to achieve the purpose of the present invention, the technical solution adopted is as follows:

[0009] A Chinese medicinal composition for preventing and treating atherosclerosis comprises the following ingredients by weight: 6-30 parts of ganoderma lucidum, 5-15 parts of zedoaria, 6-30 parts of angelica sinensis, 5-20 parts of ligusticum chuanxiong and 9-30 parts of polygonum cuspidatum.

[0010] Preferably, the raw materials include the following components by weight: 6-15 parts of Ganoderma lucidum, 5-10 parts of Curcuma zedoaria, 6-15 parts of Angelica sinensis, 5-10 parts of Ligusticum chuanxiong, and 9-15 parts of Polygonum cuspidatum.

[0011] Preferably, the raw materials include the following components by weight: 10 parts of Ganoderma lucidum, 8 parts of Curcuma zedoaria, 10 parts of Angelica sinensis, 7 parts of Ligusticum chuanxiong, and 10 parts of Polygonum cuspidatum.

[0012] The second object of the present invention is to provide a method for preparing the Chinese medicine composition, comprising the following steps:

[0013] (1) Soak Ganoderma lucidum, Curcuma zedoaria, Chinese angelica, Chuanxiong and Polygonum cuspidatum in water and boil;

[0014] (2) The decoction obtained by decoction is mixed evenly, or concentrated to obtain a thick paste, or concentrated again and then dried to obtain the product.

[0015] Preferably, the amount of water added in step (1) is 8 to 12 times the total weight of Ganoderma lucidum, Curcuma zedoaria, Angelica sinensis, Ligusticum chuanxiong and Polygonum cuspidatum, and the soaking time is 1 to 3 hours.

[0016] Preferably, the decoction in step (1) is performed 1 to 3 times, and the decoction time for each time is 20 to 60 minutes.

[0017] Preferably, the decoction in step (1) is performed twice, the first decoction lasts for 30 to 60 minutes, and the second decoction lasts for 20 to 50 minutes.

[0018] The third object of the present invention is to provide a pharmaceutical composition, the effective ingredients of which include the above-mentioned Chinese medicine composition, or the Chinese medicine composition prepared according to the above-mentioned preparation method.

[0019] Another object of the present invention is to provide a Chinese medicine composition as described above, or a Chinese medicine composition prepared according to the above preparation method, or a use of the above medicine composition in preparing a drug for treating and / or preventing atherosclerosis.

[0020] Preferably, the drug is used to reduce the blood lipid level of the body;

[0021] Preferably, the drug is used to reduce the contents of TC, TG, and LDL-C in serum and increase the content of HDL-C;

[0022] Preferably, the drug is used to inhibit inflammatory reactions, and the inflammatory factors for inhibition are TNF-α, IL-1β, and IL-12.

[0023] Preferably, the drug is used to inhibit the levels of chemokine MCP-1 and adhesion factor VCAM-1.

[0024] Preferably, the drug is used to increase the levels of catalase CAT and superoxide dismutase SOD.

[0025] Preferably, the drug is used to reduce the expression level of matrix metalloproteinase MMPs-9.

[0026] In some embodiments of the present invention, the traditional Chinese medicine composition, pharmaceutical composition, or drug can be prepared into dosage forms such as pills, capsules, granules, oral liquids, powders, tablets, lozenges, troches, etc., and suitable pharmaceutical carriers in the art can be selected according to different dosage forms.

[0027] The pharmaceutical carriers used can be solids, liquids, or gases. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil, and water. Examples of gas carriers include carbon dioxide and nitrogen.

[0028] When preparing the composition in oral dosage form, any medium that meets the requirements of pharmaceutical preparations can be used. For example, water, ethanol, oil, alcohol, flavoring agents, preservatives, coloring agents, etc. can be used to form oral liquid preparations such as suspensions and solutions; while carriers such as starch, sugars, microcrystalline cellulose, diluents, granulating agents, emulsifiers, lubricants, binders, and disintegrants can be used to form oral solid preparations such as powders, capsules, and tablets. Due to their easy administration, tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers. Standard aqueous or non-aqueous techniques can be selected for coating the tablets.

[0029] Tablets containing the Chinese medicinal composition of the present invention can be prepared by tabletting or molding, and one or more auxiliary components or adjuvants can be optionally used. The active ingredient can be tabletted in a freely flowing form (such as powder or granules) in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, surface active substance or dispersant to prepare the tablets. Molded tablets can be molded in a suitable machine, that is, a powdery mixture moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of the active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of the active ingredient. For example, a preparation intended for oral administration to humans may contain about 0.5 mg to about 5 g of the active drug, mixed with a suitable and convenient pharmaceutical carrier, which may account for about 5% to 95% of the total composition. The unit dosage form usually contains about 1 mg to about 2 g of the active ingredient, usually 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg.

[0030] The composition of the present invention can be in a suitable form for a transdermal delivery device. These formulations can be prepared using the Chinese medicinal composition of the present invention by conventional processing methods. For example, by mixing a hydrophilic material and water, and about 5 wt% to about 10 wt% of the compound, a cream or ointment with the desired consistency is prepared.

[0031] In addition to the above carrier components, the above pharmaceutical preparations may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, binders, surface active agents, thickening agents, lubricants, preservatives (including antioxidants), etc. In addition, other excipients can be added, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, coloring agents and flavoring agents, etc. The components containing the Chinese medicinal composition of the present invention can also be prepared in the form of powder or concentrate.

[0032] Terms and statements of the present invention:

[0033] The articles "a", "an" and "the": Unless otherwise explicitly limited to one (kind) of object, they include plural objects.

[0034] Numeric ranges Unless otherwise explicitly indicated, all ranges or ratios disclosed in the present invention will be understood to include any and all sub-ranges or sub-ratios contained therein. For example, the stated range or ratio of 1 to 30 should be considered to be included between the minimum value of 1 and the maximum value of 30, and any sub-range or sub-ratio, integer, decimal or sub-range or sub-ratio composed of integers or decimals including the end points.

[0035] The terms "comprising", "including", "having", "possessing", "may", "containing" and their variants are open - connecting words or terms whose meaning does not exclude the possibility of additional components or structures.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0037] (1) The key pathogenesis of AS can be summarized as "deficiency of blood vessels - accumulation of masses and stasis obstruction". Therefore, the key to the treatment method lies in regulating blood vessels, reducing lipid and resolving turbidity, and eliminating masses and removing stasis. "Vessels are the residence of blood, all blood pertains to the heart, and those who regulate vessels must first tonify the heart and nourish blood" and "vessels function by being unobstructed". The present invention is based on this.

[0038] (2) The traditional Chinese medicine composition of the present invention comprises five herbs: Ganoderma lucidum, Rhizoma zedoariae, Angelica sinensis, Ligusticum wallichii, and Polygonum cuspidatum. In the formula, Ganoderma lucidum is sweet in taste and neutral in nature, entering the heart, liver, and lung meridians, which can benefit qi and blood, soothe the mind, and is especially good at protecting the spirit, nourishing essence, and nourishing blood vessels; Rhizoma zedoariae is pungent in taste and strong in nature, specializing in promoting blood circulation within qi, mainly for breaking up stagnation and eliminating hard masses, removing accumulations and lumps. The two herbs, Ganoderma lucidum and Rhizoma zedoariae, combine sweetness and pungency, integrating dredging and nourishing, removing stasis accumulation during the regulation of blood vessels, with both dredging and tonifying, and together they are the monarch herbs; Angelica sinensis mainly enters the heart meridian, its taste is sweet and heavy, specializing in nourishing blood, and its qi is light and pungent, capable of promoting blood circulation. It has movement in tonifying and tonifying in movement, and when assisted in tonifying, it tonifies, and when assisted in attacking, it dredges; Ligusticum wallichii is pungent and warm, entering the jueyin qi aspect of the hand and foot, promoting qi and blood circulation, and the pathogen will disperse by itself. Therefore, it can break up stasis accumulation, dredge blood vessels, relieve qi stagnation, drain pus and reduce swelling, and promote blood circulation and dredge the meridians. The two herbs harmonize blood and dredge vessels, serving as ministerial herbs, which can not only assist the monarch herbs in regulating blood vessels but also clear and transform masses in the blood vessels; Polygonum cuspidatum is slightly bitter and slightly cold, which can clear and transform the heat toxin accumulated in the blood vessels and can also lead the pathogen outwards, serving as an assistant herb. The whole formula integrates dredging and nourishing, with both dredging and tonifying, removing stasis and turbidity in the blood vessels during the regulation of blood vessels, clearing and transforming masses in the blood vessels, thus having the effects of harmonizing vessels, reducing lipid, removing stasis, and eliminating masses, and having a significant therapeutic and / or preventive effect on atherosclerosis.

[0039] (3) The traditional Chinese medicine composition of the present invention has a concise compatibility and strong efficacy. Through pharmacological experimental research, it is found that the formula of the present invention can significantly reduce the blood lipid level, reduce the contents of TC, TG, and LDL - C in the serum and increase the content of HDL - C; it can significantly inhibit the levels of inflammatory factors TNF - α, IL - 1β, and IL - 12, inhibit the levels of chemotactic factor MCP - 1 and adhesion factor VCAM - 1, and reduce the expression level of matrix metalloproteinase MMPs - 9, reduce the aortic plaque area, and increase the levels of CAT and SOD to protect blood vessels. Thus, it has a significant therapeutic and / or preventive effect on atherosclerosis. Compared with the positive drug atorvastatin calcium, it has a quite strong therapeutic effect. Compared with most of the current formulas mainly composed of resolving phlegm and dredging collaterals, promoting blood circulation and reducing turbidity, the index data shows significant statistical differences. Description of the Drawings

[0040] Figure 1This is a representative image of the gross staining of oil red O on the entire length of the mouse aorta in the experimental study of the present invention, including the anatomical conditions visible to the naked eye and data comparison.

[0041] Figure 2 This is the Masson staining diagram (200X) of the aortic root of mice in each group in the experimental study of the present invention.

[0042] Figure 3 This is the level of blood lipids of mice in each group in the experimental study of the present invention, including the content comparison of four indicators: TC, TG, LDL-C, and HDL-C.

[0043] Figure 4 This is the expression of inflammatory factors in mice in each group in the experimental study of the present invention, including the secretion and expression of TNF-α, IL-10, and IL-1β.

[0044] Figure 5 This is the chemokine MCP-1 and adhesion factor VCAM-1 in the serum of mice in each group in the experimental study of the present invention.

[0045] Figure 6 This is the SOD activity value and CAT activity value in the serum of mice in each group in the experimental study of the present invention.

[0046] Figure 7 This is the immunofluorescence staining result of MMP-9 in mice in each group in the experimental study of the present invention. The upper left figure is the control group, the upper right figure is the model group, the lower left figure is the medium-dose group, and the lower right figure is the positive drug group. The scale in the figure is 100 μm.

[0047] Figure 8 This is the result diagram of the expression level of matrix metalloproteinase MMP-9 in mice in each group in the experimental study of the present invention. Detailed implementation manners

[0048] The following further describes the present invention in combination with the detailed implementation manners. The following raw materials are all commercially available conventional raw materials.

[0049] Term explanation:

[0050] AS: Atherosclerosis.

[0051] TC: Total cholesterol.

[0052] TG: Triglyceride.

[0053] LDL-C: Low-density lipoprotein cholesterol.

[0054] HDL-C: High-density lipoprotein cholesterol.

[0055] MCP-1: Monocyte chemoattractant protein-1.

[0056] VCAM-1: Vascular Cell Adhesion Molecule-1.

[0057] TNF-α: Tumor Necrosis Factor-α.

[0058] IL-1β: Interleukin-1β.

[0059] IL-12: Interleukin-12.

[0060] SOD: Superoxide Dismutase.

[0061] CAT: Catalase.

[0062] MMP-9: Matrix Metalloproteinase-9.

[0063] Example 1

[0064] This example provides a traditional Chinese medicine composition for treating and / or preventing atherosclerosis. Calculated by weight parts, its raw materials are composed of the following components: 10 parts of Ganoderma lucidum, 8 parts of Curcuma zedoaria, 10 parts of Angelica sinensis, 7 parts of Ligusticum chuanxiong, and 10 parts of Polygonum cuspidatum.

[0065] The preparation method is as follows:

[0066] (1) Soak Ganoderma lucidum, Curcuma zedoaria, Angelica sinensis, Ligusticum chuanxiong, and Polygonum cuspidatum in 10 times the amount of water for 1 h, and decoct twice. The first decoction is for 50 min, and the second decoction is for 30 min;

[0067] (2) Mix the decoction obtained from decocting, and concentrate to obtain a thick paste.

[0068] Example 2

[0069] A traditional Chinese medicine composition for treating and / or preventing atherosclerosis, calculated by weight parts, is composed of the following components: 6 parts of Ganoderma lucidum, 5 parts of Curcuma zedoaria, 6 parts of Angelica sinensis, 5 parts of Ligusticum chuanxiong, and 9 parts of Polygonum cuspidatum.

[0070] The preparation method is as follows:

[0071] (1) Soak Ganoderma lucidum, Curcuma zedoaria, Angelica sinensis, Ligusticum chuanxiong, and Polygonum cuspidatum in 8 times the amount of water for 3 h, and decoct three times. The decoction time for each time is 20 min;

[0072] (2) Mix the decoction obtained from decocting, and concentrate to obtain a thick paste.

[0073] Example 3

[0074] A traditional Chinese medicine composition for treating and / or preventing atherosclerosis, calculated by weight parts, is composed of the following components: 30 parts of Ganoderma lucidum, 15 parts of Curcuma zedoaria, 30 parts of Angelica sinensis, 20 parts of Ligusticum chuanxiong, and 30 parts of Polygonum cuspidatum.

[0075] The preparation method is as follows:

[0076] (1) Soak Ganoderma lucidum, Curcuma zedoaria, Angelica sinensis, Ligusticum wallichii, and Polygonum cuspidatum in 12 times the amount of water for 1 h, and decoct once for 60 min.

[0077] (2) Mix the decoction obtained from the decoction, and then concentrate and dry it to obtain the product.

[0078] Control Example

[0079] The formula of this control example by weight parts is: 10 parts of Allium macrostemon Bunge, 10 parts of Curcuma zedoaria, 10 parts of Coptis chinensis Franch, 15 parts of Paeonia lactiflora Pall, and 10 parts of Citrus reticulata Blanco.

[0080] The difference between the control example and Examples 1 - 3 of the present invention is that: the main efficacy of this formula is to resolve phlegm and dredge collaterals, activate blood circulation and reduce turbidity, which has a great similarity with most current traditional Chinese medicine applications, that is, the accumulation in the blood vessels is mainly guided by "resolving phlegm, reducing lipid, detoxifying, and activating blood circulation", which is quite different from the formulation idea of "combining dredging with nourishing and combining dredging and tonifying" of the traditional Chinese medicine composition of the present invention.

[0081] The preparation method is as follows:

[0082] (1) Soak Allium macrostemon Bunge, Curcuma zedoaria, Coptis chinensis Franch, Paeonia lactiflora Pall, and Citrus reticulata Blanco in 10 times the amount of water for 1 h, and decoct twice. The first decoction is for 50 min, and the second decoction is for 30 min.

[0083] (2) Mix the decoction obtained from the decoction, and concentrate to obtain a thick paste.

[0084] I. Pharmacodynamic Experimental Research

[0085] 1. Materials and Methods

[0086] 1.1 Experimental Animals

[0087] Control Group: SPF - grade male C57BL / 6 mice, Experimental Group: male ApoE mice with C57BL / 6 background - / - The mice are 8 - 9 weeks old and weigh 20 - 22 g. They are purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the license number is SYXK(Beijing)2019 - 0010. The experimental animals are raised in an SPF - grade experimental animal center. The animal environment is at a temperature of 20℃ - 26℃, a relative humidity of 50% - 70%, and the lighting time is 12 h / d (7:00 - 19:00). They can freely eat and drink water.

[0088] 1.2 Experimental Materials

[0089] High - fat diet: 78.6% basal diet + 10% lard + 1.0% cholesterol + 10.0% egg yolk powder + 0.4% bile salt, provided by Beijing Huafukang Biotechnology Co., Ltd.

[0090] Test drugs: Traditional Chinese medicine compositions prepared in Example 1 (low, medium, and high doses) and Comparative Example

[0091] Positive control drug: Atorvastatin Calcium Tablets (trade name: Lipitor, 10 mg, Pfizer Pharmaceuticals Limited).

[0092] 1.3 Experimental reagents

[0093] The main reagents used in the present invention are shown in Table 1 below:

[0094] Table 1

[0095]

[0096] Note: Shanghai Enzyme-linked refers to Shanghai Enzyme-linked Biotechnology Co., Ltd.

[0097] 1.4 Experimental instruments

[0098] The main experimental instruments used in the present invention are shown in Table 2

[0099] Table 2

[0100] Instrument Model Manufacturer Microplate Reader DR-200Bs Diatek Inverted Microscope IX51 OLYMPUS Tabletop Centrifuge TGL-16c Shanghai Anting Scientific Instrument Factory Refrigerated Centrifuge TGL-16 Hunan Xiangyi Laboratory Instruments Vortex Mixer TYXH-II Tianyue Electronics Ordinary Optical Microscope CX21 OLYMPUS Inverted White Light / Fluorescence Photography Microscope IX51 OLYMPUS Upright White Light Photography Microscope CX31 OLYMPUS Imaging System MicroPublisher Q-IMAGING Decolorizing Shaker WD-9405A Beijing Liuyi Instrument Factory

[0101] 2. Experimental methods

[0102] The experimental animals were divided into 6 groups, namely: normal group (C57BL / 6J mice), model group (ApoE - / - mice), low-dose group (ApoE - / - mice + 3.4 g crude drug / kg / d of the traditional Chinese medicine composition in Example 1), medium-dose group (ApoE - / - mice + 6.8 g crude drug / kg / d of the traditional Chinese medicine composition in Example 1), high-dose group (ApoE - / - mice + 13.6 g crude drug / kg / d of the traditional Chinese medicine composition in Example 1), positive drug group (ApoE - / - mice + 20 mg / kg / d atorvastatin calcium), and comparative example group (ApoE - / - mice + 6.8 g crude drug / kg / d of the traditional Chinese medicine composition in the comparative example), as shown in Table 3 specifically

[0103] Grouping and administration of mice

[0104]

[0105] After 1 week of adaptive feeding, the mice were fed a high-fat diet for 8 weeks. After the successful establishment of the high-fat model, the drugs were administered, and the administration period was 9 weeks

[0106] After the administration was completed, the mice were sacrificed by enucleation of the eyeballs to collect blood, which was allowed to stand at room temperature for 2 h and then centrifuged at 2500 rpm for 15 min at 4°C to separate the serum. The abdominal aorta was dissected, and a part was fixed with 4% tissue fixative, and a part was rapidly frozen with liquid nitrogen and then stored at -80°C for detection of relevant indicators.

[0107] The experimental data were expressed as Mean±SD. All experimental data were statistically analyzed using SPSS software. One-way ANOVA was used for analysis between groups of mice, and P<0.05 was considered statistically significant.

[0108] 3. Experimental results

[0109] 3.1 Oil red staining results of arterial plaques

[0110] As the target pathology, the ultimate focus of anti-AS drugs is the impact on the size / area of plaques in arteries. Regardless of the molecular biological pathway, the ultimate beneficial index for this disease is the change in the size / area of plaques in blood vessels.

[0111] As Figure 1 shown, there were no plaques in the arteries of mice in the normal group, indicating that even under the feeding of high-fat diet, C57BL / 6 mice as the blank control group would not generate atherosclerotic plaques.

[0112] Compared with the normal group, there were a large number of atherosclerotic plaque changes in the arteries of mice in the model group.

[0113] Compared with the model group, atorvastatin in the positive drug group could significantly reduce the plaque area.

[0114] Compared with the model group, the medium and high doses of the traditional Chinese medicine composition in Example 1 could reduce the plaque area along the entire length of the aorta in mice. Among them, the high dose had a more obvious reduction in plaque area than the low dose.

[0115] There was no obvious change in the comparison between the comparative example group and the model group.

[0116] 3.2 Masson staining results of the aorta

[0117] As Figure 2 shown, the structure of the aorta in mice in the normal group was dense, and the collagen fibers were arranged neatly.

[0118] Compared with the normal group, the collagen at the basal part of the blood vessels in mice in the model group was loose and there was a fracture phenomenon.

[0119] Compared with the model group, after administration of the traditional Chinese medicine composition in Example, the looseness and disordered arrangement of the blood vessel tissue at the basal part of the blood vessels in mice were improved; the collagen fiber content in the high-dose group increased significantly.

[0120] There was no significant difference when comparing the comparative example with the control group.

[0121] 3.3 Changes in blood lipid indexes of mice in each group

[0122] The results of blood lipid changes in mice in each group are shown in Table 4, Figure 3 as shown. After ApoE - / - mice were fed with a high-fat diet, typical manifestations of AS occurred, as seen in the model group. When compared with the control group, TC, TG, and LDL-C increased significantly (P < 0.01), while HDL-C decreased significantly (P < 0.01); after administration of the traditional Chinese medicine in the examples of the present invention, when compared with the model group, TC and LDL-C in the low-dose, medium-dose, and high-dose groups decreased significantly (P < 0.01), while HDL-C increased significantly (P < 0.05, P < 0.01). Although TG in the low-dose group had a downward trend, there was no significant difference; after administration of the positive drug, all indexes of AS mice showed significant changes (P < 0.01); after administration of the traditional Chinese medicine in the comparative example, there were statistical differences in all indexes when compared with the model group, indicating that this combined formula can have a significant effect on blood lipids. As Figure 3 shown, when compared with the medium dose of the example, indexes such as TC, TG, and LDL-C in the traditional Chinese medicine group of the comparative example were significantly higher than the medium-dose level (P < 0.05, P < 0.01).

[0123] Table 4

[0124] Group TC (mmol / L) TG (mmol / L) LDL-C (mmol / L) HDL-C (mmol / L) Control Group 2.84±0.86 0.89±0.26 1.03±0.15 1.99±0.46 Model Group 14.25±1.21## 5.07±0.34## 13.81±1.86## 0.36±0.10## Low Dose Group 10.80±0.88** 4.16±0.46 9.85±0.85** 1.22±0.06* Medium Dose Group 8.84±0.40** 2.87±0.20** 8.05±1.11** 1.45±0.26* High Dose Group 6.28±0.76** 2.53±0.38** 5.96±0.81** 1.73±0.40** Positive Drug Group 5.75±0.38** 1.70±0.57** 3.41±1.13** 1.89±0.49** Comparison Group <![CDATA[11.76±1.25* △△ > <![CDATA[3.92±0.50* △ > <![CDATA[10.95±0.51* △ > 1.23±0.13*

[0125] Note: # Compared with the control group: P < 0.05; ## Compared with the control group: P < 0.01; * Compared with the model group: P < 0.05; ** Compared with the model group: P < 0.01; △ Compared with the medium-dose group: P < 0.05; △△ Compared with the medium-dose group: P < 0.01.

[0126] 3.4 Changes in indexes related to inflammatory factors of mice in each group

[0127] The detection results of inflammatory factor levels in mice in each group are shown in Table 5, Figure 4 as shown.

[0128] Compared with the control group, the expression levels of TNF-α, IL-1β, and IL-12 in the AS model group of mice were all significantly increased (P < 0.01), indicating severe vascular damage in the mice. After administration of the drug, compared with the model group, the expressions of TNF-α and IL-1β in the low-dose group of the examples were significantly decreased (P < 0.01, P < 0.05), and the expression of IL-12 also showed a downward trend, but there was no statistical difference. The expression levels of TNF-α, IL-1β, and IL-12 in the medium-dose and high-dose groups of the examples were all significantly decreased (P < 0.01). After administration of the positive drug, compared with the model group, the expression levels of TNF-α, IL-1β, and IL-12 were also significantly decreased (P < 0.01). After administration of the traditional Chinese medicine group in the comparative example, there was a significant difference compared with the model group, and compared with the medium-dose group, the expression levels of TNF-α, IL-1β, and IL-12 were significantly higher than those in the medium-dose group (P < 0.05).

[0129] Table 5

[0130] Group TNF-α (pg / mL) IL-1β (pg / mL) IL-12 (pg / mL) Control Group 53.63±11.08 33.56±13.01 5.40±1.06 Model Group 364.2±37.18## 323.5±30.42## 35.62±3.22## Low Dose Group 280.7±28.44** 254.4±35.73* 28.66±3.90 Medium Dose Group 220.8±25.24** 170.8±36.32** 20.79±0.72** High Dose Group 115.2±16.61** 79.78±17.03** 15.96±2.59** Positive Drug Group 145.0±15.35** 144.9±23.66** 10.17±2.18** Comparison Group <![CDATA[285.8±28.51* △ > <![CDATA[245.5±35.33* △ > <![CDATA[28.31±1.58* △ >

[0131] Note: # Compared with the control group: P < 0.05; ## Compared with the control group: P < 0.01; * Compared with the model group: P < 0.05; ** Compared with the model group: P < 0.01; △ Compared with the medium-dose group: P < 0.05.

[0132] 3.5 Expression differences of chemokines and adhesion factors in each group of mice

[0133] The chemokine MCP-1 can attract monocytes in the blood, make them adhere to the vascular endothelium and penetrate into the subendothelium, and then ingest cholesterol to form foam cells, thus triggering and aggravating the process of AS; the vascular adhesion factor VCAM-1 is one of the key markers of AS, and its high expression can promote the adhesion, migration and infiltration of leukocytes on the vascular wall, and then exacerbate the inflammatory reaction and damage of the vascular wall, thus promoting the development of the disease.

[0134] The expressions of MCP-1 and VCAM-1 in each group of mice are shown in Table 6 Figure 5 as follows.

[0135] The expressions of MCP-1 and VCAM-1 in the mice of the AS model group were both highly expressed compared with the control group (P < 0.01), indicating that the environment of atherosclerosis in the model group mice was severe; after administration, the expression levels of MCP-1 and VCAM-1 in the low-dose group, medium-dose group, and high-dose group of the examples were significantly decreased compared with the model group (P < 0.01, P < 0.05); after administration of the positive drug, compared with the model group, the expressions of both indicators were significantly decreased (P < 0.01); after administration of the traditional Chinese medicine in the comparative example, there was a statistically significant difference in the expression compared with the model group, but compared with the medium-dose group, the expression levels of MCP-1 and VCAM-1 were both significantly increased (P < 0.01).

[0136] Table 6

[0137] Group MCP-1 (pg / mL) VCAM-1 (pg / mL) Control Group 46.10±5.76 513.9±62.78 Model Group 557.0±47.7## 5439±509.8## Low Dose Group 396.3±1.41** 4296±227.4* Medium Dose Group 300.3±7.84** 3408±497.8** High Dose Group 167.3±27.5** 2337±61.27** Positive Drug Group 247.9±38.6** 1503±232.0** Comparison Group <![CDATA[467.4±17.8* △△ > <![CDATA[4472±133.5* △△ >

[0138] Note: # Compared with the control group: P < 0.05; ## Compared with the control group: P < 0.01; * Compared with the model group: P < 0.05; ** Compared with the model group: P < 0.01; △△ Compared with the medium-dose group: P < 0.01.

[0139] 3.6 Expression differences of antioxidant enzymes in each group of mice

[0140] The antioxidant system is a key protective mechanism that helps the body resist the attack of free radicals. Superoxide dismutase (SOD) reduces the number of superoxide anion free radicals, thereby reducing the damage of free radicals to cells; catalase (CAT) rapidly scavenges hydrogen peroxide to prevent the accumulation of free radicals and damage to cells. Such indicators can indicate whether the traditional Chinese medicine composition of the present invention has a protective effect on blood vessels.

[0141] The expressions of SOD and CAT in each group of mice are shown in Table 7 Figure 6 as follows

[0142] Table 7

[0143] Group SOD Activity (U / mL) CAT Activity (U / mL) Control Group 102.5±9.13 21.14±2.36 Model Group 26.02±1.96## 3.568±1.21## Low Dose Group 43.04±1.43 7.633±1.72 Medium Dose Group 54.06±9.34** 11.19±0.67** High Dose Group 70.00±10.7** 14.12±1.86** Positive Drug Group 86.30±7.45** 17.68±2.33** Comparison Group <![CDATA[33.17±4.32 △ > <![CDATA[6.853±0.75 △ >

[0144] Note: # Compared with the control group: P < 0.05; ## Compared with the control group: P < 0.01; * Compared with the model group: P < 0.05; ** Compared with the model group: P < 0.01; △ Compared with the medium-dose group: P < 0.05.

[0145] The SOD and CAT of the mice in the AS model group were significantly decreased compared with those of the control group (P < 0.01), indicating that the intravascular environment of atherosclerosis in the model group mice was poor and suffered great damage; after administration, the expression levels of SOD and CAT in the medium-dose group and high-dose group of the examples were significantly increased compared with those of the model group (P < 0.01), and there was a strong tendency of increase in the low-dose group, but there was no statistical difference; after administration of the positive drug, compared with the model group, the expressions of both indicators were significantly increased (P < 0.01); after administration of the traditional Chinese medicine in the comparative example, there was no significant difference in the expression compared with the model group, but compared with the medium-dose group, the expression levels of SOD and CAT were both low expression (P < 0.05).

[0146] 3.7 Changes of matrix metalloproteinase MMP-9 in each group of mice

[0147] Matrix metalloproteinases MMPs are considered to be an important proteolytic enzyme, which can degrade almost all protein components in the ECM and participate in atherosclerosis and plaque rupture. MMP-9 can participate in the degradation and absorption process of plaques.

[0148] The immunofluorescence staining results of MMP-9 in each group of mice are shown in Figure 7 and the results of MMP-9 expression levels are shown in Figure 8 and Table 8.

[0149] Table 8

[0150] Group MMP-9 Expression Level Control Group 1062±168.6 Model Group 6432±653.8## Medium Dose Group 4092±303.5** Positive Drug Group 3326±349.5**

[0151] Note: ## Compared with the control group: P < 0.01; * Compared with the model group: P < 0.05; ** Compared with the model group: P < 0.01.

[0152] Compared with the normal group, it was observed under the microscope that MMP-9 was expressed in foam cells, macrophages, vascular endothelial cells and vascular smooth muscle cells in the arterial plaque tissue of the mice in the AS group, especially in foam cells and macrophages in large amounts (P < 0.01), indicating that MMP-9 was activated in the AS state; after administration of the medium-dose group in the examples, compared with the model group, the expression level of MMP-9 was significantly decreased (P < 0.01); compared with the model group, the expression level of MMP-9 in the positive drug atorvastatin group was also significantly decreased (P < 0.01), and there was no obvious difference between the medium-dose group in the examples and the positive drug group.

[0153] II. Clinical case

[0154] In the clinical practice of the present invention, according to individual differences, patients clinically diagnosed with atherosclerosis and hyperlipidemia or taking the traditional Chinese medicine compositions in Example 2 or Example 3 of the present invention for several weeks, and the results showed that: the blood lipid levels and the plaque areas of atherosclerosis of the patients were significantly reduced.

[0155] 1. Wang, male, 59 years old, clinically diagnosed with hyperlipidemia accompanied by atherosclerosis, took the prescription of Example 2 for 4 months, and the blood lipid indexes before and after taking the medicine are as shown in Table 9 below.

[0156] Table 9

[0157]

[0158] Before taking the medicine, the carotid artery color Doppler ultrasound results showed that the bilateral common carotid arteries, internal carotid arteries, and external carotid arteries had natural courses, the lumen structures were clearly shown, the intima was rough and thickened, the thickest parts of the intima-media of the right and left common carotid arteries were about 0.10 cm and 0.15 cm respectively, and an isoechoic plaque with a size of about 0.64 * 0.21 cm was visible at the enlargement of the right common carotid artery. CDFI: Color blood flow signal filling defect was visible at the thickened intima and plaque, the artery showed pulsatile color blood flow signal, and PW: The arterial blood flow spectrum showed no obvious abnormality.

[0159] After taking the medicine, the carotid artery color Doppler ultrasound results showed that the bilateral common carotid arteries, internal carotid arteries, and external carotid arteries had natural courses, the lumen structures were clearly shown, the intima was rough and thickened, the thickest parts of the intima-media of the right and left common carotid arteries were about 0.11 cm and 0.11 cm respectively, and an isoechoic plaque with a size of about 0.60 * 0.18 cm was visible at the enlargement of the right common carotid artery. CDFI: Color blood flow signal filling defect was visible at the thickened intima and plaque, the artery showed pulsatile color blood flow signal, and PW: The arterial blood flow spectrum showed no obvious abnormality.

[0160] 2. Xue, female, 58 years old, clinically diagnosed with hyperlipidemia accompanied by atherosclerosis, took the prescription of Example 3 for 3 months, and the blood lipid-related indexes before and after taking the medicine are as shown in Table 10 below.

[0161] Table 10

[0162]

[0163] Findings of carotid artery color Doppler ultrasound before taking the medicine: The bilateral common carotid arteries, internal carotid arteries, and external carotid arteries had natural courses, the lumen structures were clearly shown, the intima was rough and slightly thickened, the thickest parts of the intima-media of the right and left common carotid arteries were about 0.11 cm and 0.11 cm respectively, and a hyperechoic plaque with a size of about 0.21 * 0.09 cm was visible at the enlargement of the left common carotid artery. CDFI: Color blood flow signal filling defect was visible at the thickened intima and plaque, the artery showed pulsatile color blood flow signal, and PW: The arterial blood flow spectrum showed no obvious abnormality.

[0164] Findings of carotid artery color ultrasound after taking medicine: The bilateral common carotid arteries, internal carotid arteries, and external carotid arteries have natural courses, clear lumen structures, rough and thickened intima. The thickest parts of the intima-media of the right and left common carotid arteries are about 0.10 cm and 0.10 cm respectively. CDFI shows that there are filling defects of color blood flow signals at the thickened intima, and the artery shows pulsatile color blood flow signals. PW shows that there are no obvious abnormalities in the arterial blood flow spectrum.

[0165] 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. Any equivalent implementation or change without departing from the present invention shall be included within the scope of the technical solution of the present invention.

Claims

1. A Chinese medicine composition for preventing and treating atherosclerosis, characterized in that: The raw materials include the following components by weight: 6-30 parts of ganoderma lucidum, 5-15 parts of zedoaria, 6-30 parts of angelica sinensis, 5-20 parts of chuanxiong and 9-30 parts of polygonum cuspidatum.

2. The Chinese medicine composition according to claim 1, characterized in that: The raw materials include the following components by weight: 6-15 parts of ganoderma lucidum, 5-10 parts of zedoaria, 6-15 parts of angelica sinensis, 5-10 parts of chuanxiong and 9-15 parts of polygonum cuspidatum.

3. The Chinese medicine composition according to claim 1, characterized in that The raw materials include the following components by weight: 10 parts of ganoderma lucidum, 8 parts of zedoary turmeric, 10 parts of angelica sinensis, 7 parts of ligusticum chuanxiong and 10 parts of polygonum cuspidatum.

4. A method for preparing the Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The steps include: (1) Soak Ganoderma lucidum, Curcuma zedoaria, Chinese angelica, Chuanxiong and Polygonum cuspidatum in water and boil; (2) The decoction obtained by decoction is mixed evenly, or concentrated to obtain a thick paste, or concentrated and dried to obtain the product.

5. The preparation method according to claim 4, characterized in that: The amount of water added in step (1) is 8 to 12 times the total weight of Ganoderma lucidum, Curcuma zedoaria, Angelica sinensis, Ligusticum chuanxiong and Polygonum cuspidatum, and the soaking time is 1 to 3 hours.

6. The preparation method according to claim 4, characterized in that: The decoction is performed 1 to 3 times in step (1), and the decoction time for each time is 20 to 60 minutes.

7. The preparation method according to claim 6, characterized in that: The decoction in step (1) is performed twice, the first decoction is 30 to 60 minutes, and the second decoction is 20 to 50 minutes.

8. A pharmaceutical composition, characterized in that The effective ingredients include the Chinese medicine composition according to any one of claims 1 to 3, or the Chinese medicine composition prepared by the preparation method according to any one of claims 4 to 7.

9. Use of the Chinese medicine composition according to any one of claims 1 to 3, or the Chinese medicine composition prepared by the preparation method according to any one of claims 4 to 7, or the pharmaceutical composition according to claim 8 in the preparation of a drug for treating and / or preventing atherosclerosis.

10. The use according to claim 9, characterized in that: The drug is used to reduce the body's blood lipid level, and / or reduce the content of TC, TG, and LDL-C in serum, increase the content of HDL-C, and / or inhibit the level of inflammatory factors, and / or inhibit the level of chemokine MCP-1 and adhesion factor VCAM-1, and / or increase the level of catalase CAT and superoxide dismutase SOD, and / or reduce the expression of matrix metalloproteinase MMPs-9.

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