A traditional Chinese medicine composition for treating calcific aortic valve disease, its preparation method and uses

Abstract

The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for treating calcific aortic valve disease, a preparation method thereof, and uses thereof. The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 10 - 50 parts of astragalus membranaceus, 5 - 40 parts of radix paeoniae rubra, 5 - 40 parts of stir-fried atractylodes macrocephala, 1 - 20 parts of polygonatum sibiricum processed with wine, and 1 - 20 parts of zedoary turmeric processed with vinegar. The present invention intervenes in calcific aortic valve disease by the method of supplementing qi and activating blood circulation, strengthening the healthy qi and eliminating stasis, improves the local or diffuse echo enhancement of the valve under ultrasound, thickening of the valve, stenosis of the aortic valve orifice area, reduces the peak flow velocity, and reduces the degree of aortic valve calcification, and is used for treating symptoms such as chest tightness, chest pain, shortness of breath, fatigue, and palpitations caused by the accumulation of masses due to qi deficiency and blood stasis in patients with aortic valve calcification, and is particularly suitable for those with calcific aortic valve disease showing the above symptoms.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition for treating calcific aortic valve disease, a preparation method and use thereof. Background Art

[0002] Calcific aortic valve disease (CAVD) is one of the most common valvular heart diseases. Its disease spectrum covers diseases ranging from aortic valve sclerosis to calcific aortic valve stenosis that can cause severe left ventricular outflow tract obstruction. Among them, aortic valve sclerosis occurs before aortic valve stenosis. About 9% of patients with aortic valve sclerosis will develop aortic valve stenosis within 5 years. Population-based cardiovascular health studies have shown that aortic valve stenosis is a progressive disease. Every year, 1%-2% of aortic valve sclerosis will progress to aortic valve stenosis, and the prevalence increases with age. The average prevalence in people aged 50-59 is 0.2%, and the prevalence in people aged 80-89 can increase to 9.8%. Among them, 75% of patients will experience heart failure and receive valve replacement surgery, or die within 2-5 years. Patients are usually asymptomatic in the early stage. When symptoms appear, the prognosis is usually poor. It is a major public health problem that needs to be solved in contemporary society.

[0003] Currently, the treatment options for calcific aortic valve disease are relatively limited. The main interventions are surgical aortic valve replacement (SAVR), transcatheter aortic valve implantation (TAVI), or transcatheter aortic valve replacement (TAVR), which can reduce the mortality rate of CAVD to a certain extent. However, they carry high risks, high costs, and some patients miss the best treatment period due to advanced age or pre-existing other diseases and are unable to tolerate surgery. Pathophysiological studies have pointed out that the early changes in CAVD share a common disease progression process with AS. For some time, lipid-lowering therapy has become the focus of research on CAVD intervention. However, recent clinical research results at home and abroad have shown that statin lipid-lowering drugs are not ideal for the treatment of CAVD, and even relatively ineffective. In addition, the latest epidemiological studies have pointed out that with the increase in human lifespan and the influence of diet and work environment, the disease burden of CAVD shows a continuous growth trend in the elderly population. It is estimated that by 2050, the number of patients in the United States and Europe who need SAVR or TAVI will triple. Due to the differences in the accessibility of surgical treatment globally, the disease risks cannot be ignored. It is urgent to develop drug treatment regimens that can effectively delay or prevent the progression of CAVD.

[0004] With thousands of years of historical precipitation, traditional Chinese medicine (TCM) has rich experience in the treatment of cardiovascular diseases. Guided by TCM theory, TCM starts from the overall concept, dialectically grasps the evolution law of diseases, and has obvious advantages in the prevention and treatment of complex diseases. Moreover, traditional Chinese medicine has the treatment characteristics of multiple components, multiple pathways, multiple levels, multiple targets and small adverse reactions, which has great value for in-depth research and application. There is no record of the disease name of "calcific aortic valve disease" in TCM literature. According to its pathogenesis and clinical symptoms, it can be classified into the categories of "chest pain, palpitation, shortness of breath, edema, syncope" and other diseases in TCM. In its mild form, only symptoms such as fatigue may be seen, and the clinical symptoms are not prominent; in its severe form, symptoms such as dyspnea, syncope, chest pain, general fatigue, edema, and palpitation may occur. The disease location syndrome factors are mostly the heart, lungs, and kidneys; the distribution of the disease nature syndrome factors for the first three are phlegm, qi deficiency, and yang deficiency respectively. The Inner Canon of Huangdi states: "At the age of sixty, the heart qi begins to decline, and the qi and blood become sluggish" - qi deficiency is a physiological change in the process of human aging, and heart qi deficiency is an inducing factor for the onset of the disease. Deficiency leads to excess syndromes such as blood stasis, retention of phlegm, fluid, and dampness. Therefore, the basic pathogenesis of CAVD in the theory of integrated traditional Chinese and Western medicine is as follows: improper diet, addiction to alcohol and tobacco, aging, qi decline, and weakness of the spleen and stomach, resulting in dysfunction of transportation and transformation, stagnation of water, dampness, and body fluids, accumulation of phlegm, blockage of the collaterals by phlegm, resulting in qi and blood stasis, which can cause this disease; emotional injury leads to disorders of the body's zang-fu functions, abnormal qi and blood circulation, the heart and liver qi and fire boiling body fluids and nutrient blood, fire burning the collaterals, and over time, toxin accumulation and blockage of the meridians and channels, resulting in blood stasis and the formation of masses, thus causing the disease; in addition, due to old age and physical weakness, the kidney yang gradually declines, the kidney yin is insufficient, the decline of kidney yang leads to unfavorable qi transformation, the deficiency of kidney yin leads to inability to moisten, the distribution of qi, blood, and body fluids is disordered, and the decline of kidney qi over time leads to weakness of the vital qi, which is insufficient to promote blood circulation, and the stagnation of blood circulation leads to blood stasis, exacerbating the formation of masses.

[0005] Discussions on relevant symptoms and pathogenesis can be found in ancient TCM classics. For example, On Chest Bi, Heart Pain, and Shortness of Breath in Synopsis of the Golden Chamber states: "When taking the pulse, one should pay attention to excess and deficiency. Yang is weak and yin is stringy, which means chest bi and pain. The reason for this is that it is extremely deficient. Now, yang deficiency is known to be in the upper jiao. Therefore, the chest bi and heart pain are due to the stringy yin." It points out that the cause is "yang deficiency and yin stringiness" - deficiency of vital qi is the root cause; Plain Questions: Treatise on Bi Syndromes states: "For those with heart bi, the vessels are not unobstructed", believing that blood stasis and unsmooth vessels are the main pathogenesis. To sum up, deficiency of vital qi, mutual binding of phlegm and stasis, and accumulation of masses are the main TCM pathogenesis of CAVD.

[0006] In view of this, the present invention treats CAVD by the method of supplementing qi and activating blood circulation, strengthening the healthy qi and eliminating masses, improves the local or diffuse echo enhancement of the valve under ultrasound, thickening of the valve, stenosis of the aortic valve orifice area, reduces the peak flow velocity, and reduces the degree of aortic valve calcification, so as to develop drugs applicable to CAVD with symptoms such as chest tightness and shortness of breath to serve clinical practice. Summary of the Invention

[0007] To solve the problem of the shortage of traditional Chinese medicine prescriptions that can effectively treat CAVD for clinical use, the present invention provides a novel traditional Chinese medicine composition for treating calcific aortic valve disease, its preparation method and uses.

[0008] Specifically, the present invention is achieved through the following technical solutions in several aspects:

[0009] In the first aspect, the present invention provides a traditional Chinese medicine composition for treating calcific aortic valve disease, and the traditional Chinese medicine composition is made from the following raw materials by weight: 10 - 50 parts of Astragalus membranaceus, 5 - 40 parts of Paeonia lactiflora Pall., 5 - 40 parts of Atractylodes macrocephala Koidz. stir-fried, 1 - 20 parts of Polygonatum sibiricum Delar. ex Redoute processed with wine, 1 - 20 parts of Rhizoma Curcumae processed with vinegar.

[0010] In a preferred embodiment, the traditional Chinese medicine composition is made from the following raw materials by weight: 15 - 40 parts of Astragalus membranaceus, 10 - 30 parts of Paeonia lactiflora Pall., 10 - 30 parts of Atractylodes macrocephala Koidz. stir-fried, 5 - 15 parts of Polygonatum sibiricum Delar. ex Redoute processed with wine, 5 - 15 parts of Rhizoma Curcumae processed with vinegar.

[0011] In a more preferred embodiment, the traditional Chinese medicine composition is made from the following raw materials by weight: 20 - 30 parts of Astragalus membranaceus, 12 - 20 parts of Paeonia lactiflora Pall., 15 - 20 parts of Atractylodes macrocephala Koidz. stir-fried, 8 - 10 parts of Polygonatum sibiricum Delar. ex Redoute processed with wine, 8 - 10 parts of Rhizoma Curcumae processed with vinegar.

[0012] In a most preferred embodiment, the traditional Chinese medicine composition is made from the following raw materials by weight: 30 parts of Astragalus membranaceus, 20 parts of Paeonia lactiflora Pall., 20 parts of Atractylodes macrocephala Koidz. stir-fried, 10 parts of Polygonatum sibiricum Delar. ex Redoute processed with wine, 10 parts of Rhizoma Curcumae processed with vinegar.

[0013] In another most preferred embodiment, the traditional Chinese medicine composition is made from the following raw materials by weight: 20 parts of Astragalus membranaceus, 12 parts of Paeonia lactiflora Pall., 15 parts of Atractylodes macrocephala Koidz. stir-fried, 8 parts of Polygonatum sibiricum Delar. ex Redoute processed with wine, 8 parts of Rhizoma Curcumae processed with vinegar.

[0014] In another preferred embodiment, the traditional Chinese medicine composition has the effects of supplementing qi and activating blood circulation, strengthening the healthy qi and eliminating stasis.

[0015] In the second aspect, the drug of the present invention can be prepared by conventional preparation methods in the art, and pharmaceutically acceptable excipients can be optionally added or not added. As long as the traditional Chinese medicine raw materials are 10 - 50 parts of Astragalus membranaceus, 5 - 40 parts of Paeonia lactiflora Pall., 5 - 40 parts of Atractylodes macrocephala Koidz. stir-fried, 1 - 20 parts of Polygonatum sibiricum Delar. ex Redoute processed with wine, 1 - 20 parts of Rhizoma Curcumae processed with vinegar, regardless of various changes in its preparation method, it falls within the scope of the present invention. Its drug forms include solid forms, semi-solid forms or liquid forms, etc.; specifically, including but not limited to cut crude drugs, decoctions, powders, oral liquids, ointments, granules, tablets or capsules.

[0016] In another most preferred embodiment, the present invention provides a method for preparing the traditional Chinese medicine composition described in the first aspect above. The preparation method includes the following steps: weighing each raw material medicine according to the weight parts described in the first aspect above, extracting, and concentrating.

[0017] Preferably, in the above preparation method, the traditional Chinese medicine composition of the present invention may further include a pretreatment step.

[0018] Preferably, it includes weighing each raw material medicine, pretreatment, extraction, and concentration.

[0019] Preferably, the pretreatment step includes soaking and / or pulverizing.

[0020] Preferably, in the above preparation method, the traditional Chinese medicine composition of the present invention is prepared by a conventional method well known in the art.

[0021] Preferably, the extraction methods used in the present invention include but are not limited to decoction extraction, reflux extraction, ultrasonic extraction, supercritical extraction, etc.

[0022] Preferably, water, organic solvents commonly used in the art, or a mixture thereof can be used to extract the treated raw material medicine.

[0023] Preferably, the organic solvent is methanol, ethanol, propanol, butanol, ethyl acetate, petroleum ether, etc.

[0024] In a preferred embodiment, the number of decoction times is two. After the first decoction for 1 - 3 hours, the first filtrate is obtained by filtration. The filter residue is then decocted with water for 1 - 3 hours, and the second filtrate is obtained by filtration. The first filtrate and the second filtrate are combined.

[0025] In a more preferred embodiment, the water used for the first decoction and the second decoction is distilled water; the concentration is vacuum concentration.

[0026] In addition, as an alternative, a traditional Chinese medicine decoctor can be used to prepare the traditional Chinese medicine composition of the present invention, which specifically includes the following steps:

[0027] 1. Preparation work:

[0028] (1) Put the medicine to be processed into a special non-woven bag, tie the mouth well, put it into a container, and soak it in cold water for 30 minutes;

[0029] (2) Clean the traditional Chinese medicine decoctor, open the waste liquid discharge valve, drain the water completely, close the waste liquid discharge valve, close the liquid discharge valve, put the soaked medicine bag into the medicine pot, and add the soaked liquid and water;

[0030] (3) Water addition ratio: The amount of added water = the required amount of liquid medicine + 20%.

[0031] Calculation formula: Amount of water added = (Capacity per bag of medicine × Number of times taken daily × Number of decoctions) × 1.2.

[0032] 2. Operating steps for decocting medicine:

[0033] (1) Put traditional Chinese medicine and water into the decocting machine, cover the pot lid, tighten the handle, close the exhaust safety valve, set the decocting time and temperature, press the heating button, the machine will heat automatically, switch to slow fire automatically when reaching the set temperature, and stop heating automatically after reaching the set time;

[0034] (2) After the medicine is decocted, first open the exhaust safety valve to reduce the pressure appropriately (make the pressure drop to about 1 kg), then open the liquid discharge valve, and the liquid medicine will be injected into the medicine tank of the packaging machine through the liquid discharge hose. During the process of discharging the liquid medicine, rotate the extrusion device at the same time to squeeze out the residual liquid medicine in the medicine bag. After the liquid medicine is discharged, open the exhaust safety valve, reduce the pressure to atmospheric pressure, open the handle to take out the medicine bag, turn off the power supply, and clean for standby.

[0035] In the third aspect, the present invention provides the use of the traditional Chinese medicine composition described in the first aspect above or the traditional Chinese medicine composition prepared by the preparation method described in the second aspect above in the preparation of a medicament for treating calcific aortic valve disease.

[0036] In a preferred embodiment, the medicament treats symptoms such as chest tightness, chest pain, shortness of breath, fatigue, and palpitations caused by qi deficiency and blood stasis resulting in masses and accumulations in patients with calcific aortic valve disease.

[0037] In a more preferred embodiment, the medicament improves local or diffuse echo enhancement of the valve under ultrasound, thickening of the valve, stenosis of the aortic valve orifice area, reduces the peak flow velocity, and reduces the degree of aortic valve calcification.

[0038] In another preferred embodiment, the medicament is in solid form, semi-solid form or liquid form.

[0039] In another preferred embodiment, the medicament is in the form of cut crude drugs, decoction, powder, oral liquid, paste, granule, tablet or capsule.

[0040] In another preferred embodiment, the dosage form of the medicament is decoction or granule, tablet, oral liquid or capsule.

[0041] In another preferred embodiment, the dosage form of the medicament is decoction or granule.

[0042] Preferably, the dosage form of the medicament is granule.

[0043] The present invention has the following beneficial effects compared with the prior art:

[0044] The present invention intervenes in calcific aortic valve disease by the method of supplementing qi and activating blood circulation, strengthening the healthy qi and eliminating stasis, improving local or diffuse echo enhancement of the valve under ultrasound, valve thickening, aortic valve orifice area stenosis, reducing peak velocity, and reducing the degree of aortic valve calcification, and is used for treating symptoms such as chest tightness, chest pain, shortness of breath, fatigue, and palpitations caused by the accumulation of masses due to qi deficiency and blood stasis in patients with calcific aortic valve disease, and is particularly applicable to those with the above symptoms of calcific aortic valve disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 : Von Kossa staining results of aortic valves of mice in each group.

[0046] Figure 2 : Alizarin red staining results of aortic valves of mice in each group.

[0047] Figure 3 : RUNX2 staining results of aortic valve calcification markers in mice in each group.

[0048] Figure 4 : BMP2 staining results of aortic valve calcification markers in mice in each group. DETAILED DESCRIPTION OF THE INVENTION

[0049] The analysis of the formula of the traditional Chinese medicine composition of the present invention is as follows:

[0050] Astragalus membranaceus, the root of Astragalus membranaceus (Fisch.) Bunge var. mongholicus (Bunge) Hsiao or Astragalus membranaceus (Fisch.) Bunge of the Leguminosae family, sweet in taste, slightly warm in nature, attributing to the spleen and lung meridians, having the effects of invigorating the spleen and replenishing qi, lifting yang and raising prolapse, benefiting qi and strengthening the exterior, inducing diuresis, and promoting granulation and detoxicating. Astragalus memeranaceus(Fisch.)Bge. Var. mongholicus(Bge.)Hsiao or Astragalus membranaceus (Fisch.) Bunge A. membranaceus(Fisch.)Bge. The root of Astragalus membranaceus (Fisch.) Bunge var. mongholicus (Bunge) Hsiao or Astragalus membranaceus (Fisch.) Bunge of the Leguminosae family, sweet in taste, slightly warm in nature, attributing to the spleen and lung meridians, having the effects of invigorating the spleen and replenishing qi, lifting yang and raising prolapse, benefiting qi and strengthening the exterior, inducing diuresis, and promoting granulation and detoxicating.

[0051] Paeonia lactiflora Pall., the dried root of Paeonia lactiflora Pall. or Paeonia veitchii Lynch of the Ranunculaceae family, bitter in taste, slightly cold in nature, attributing to the liver meridian, having the effects of clearing heat and cooling blood, and dissipating stasis and relieving pain. Paeonia lactiflora Pall. or Paeonia veitchii Lynch Paeonia veitchii Lynch The dried root of Paeonia lactiflora Pall. or Paeonia veitchii Lynch of the Ranunculaceae family, bitter in taste, slightly cold in nature, attributing to the liver meridian, having the effects of clearing heat and cooling blood, and dissipating stasis and relieving pain.

[0052] Atractylodes macrocephala Koidz., the rhizome of Atractylodes macrocephala Koidz. of the Compositae family, warm in nature, sweet and bitter in taste, attributing to the spleen and stomach meridians, having the effects of strengthening the spleen, replenishing qi, drying dampness and promoting diuresis, stopping sweating, and preventing abortion. It belongs to the qi-tonifying drugs under the classification of tonic drugs. It is used to treat symptoms such as weakness of the spleen and stomach qi, loss of appetite, lassitude, flaccidity, diarrhea, phlegm retention, edema, jaundice, damp arthralgia, dysuria, dizziness, spontaneous sweating, and fetal restlessness. Atractylodes macrocephala Koidz. The rhizome of Atractylodes macrocephala Koidz. of the Compositae family, warm in nature, sweet and bitter in taste, attributing to the spleen and stomach meridians, having the effects of strengthening the spleen, replenishing qi, drying dampness and promoting diuresis, stopping sweating, and preventing abortion. It belongs to the qi-tonifying drugs under the classification of tonic drugs. It is used to treat symptoms such as weakness of the spleen and stomach qi, loss of appetite, lassitude, flaccidity, diarrhea, phlegm retention, edema, jaundice, damp arthralgia, dysuria, dizziness, spontaneous sweating, and fetal restlessness.

[0053] Polygonatum sibiricum Red., the rhizome of Polygonatum kingianum Coll. et Hemsl., Polygonatum sibiricum Red. or Polygonatum cyrtonema Hua of the Liliaceae family Polygonatum kingianum Coll.et Hemsl . Polygonatum sibiricum Red. Polygonatum sibiricum Red. or Polygonatum cyrtonema Hua Polygonatum cyrtonema HuaThe dried rhizome is stewed or steamed thoroughly by the method of stewing with wine or steaming with wine, slightly aired, cut into thick slices, and dried. It tastes sweet, is neutral in nature, and belongs to the spleen, lung, and kidney meridians. It has the effects of replenishing qi and nourishing yin, strengthening the spleen, moistening the lungs, and tonifying the kidneys. Clinically, it is commonly used for symptoms such as qi deficiency of the spleen and stomach, fatigue, yin deficiency of the stomach, insufficiency of essence and blood, soreness and weakness of the waist and knees, premature whitening of hair, etc.

[0054] Rhizoma Curcumae Aromaticae Vinegar-processed, is the rhizome of the plants Curcuma zedoaria (Christm.) Rosc., Curcuma aeruginosa Roxb.[C.zedoaria non Rosc.] Curcuma kwangsiensis S. G. Lee et C. F. Liang, Curcuma kwangsiensis S. G. Lee et C. F. Liang and Curcuma wenyujin Y. H. Chen et C. Ling Curcuma wenyujin Y. H. Chen et C. Ling in the family Zingiberaceae. It tastes pungent and bitter, is warm in nature, and belongs to the liver and spleen meridians. It has the effects of promoting qi circulation to break blood stasis and removing stasis to relieve pain.

[0055] Among them, raw Astragalus membranaceus is the monarch drug. While replenishing qi, it also has the effect of lifting, tonifying the deficiency of middle qi, relieving chest distress, shortness of breath, and fatigue caused by calcific aortic valve disease. Replenishing qi can promote the circulation of water and dampness, and the use of Astragalus membranaceus can also promote urination, relieve the symptoms of chest pain, chest distress, shortness of breath, fatigue, and palpitations caused by calcific aortic valve disease. The ministerial drug is Paeonia lactiflora Pall. var. trichocarpa (Bunge) Stern. It functions to cool blood and promote blood circulation, and can also promote urination - "The red peony root promotes urination slightly. In common prescriptions, it is used to relieve pain, and its effect is not inferior to that of Angelica sinensis" (Compendium of Materia Medica Collected Annotations). Modern research further points out that Paeonia lactiflora Pall. var. trichocarpa (Bunge) Stern. can reduce plasma peroxidized lipids, improve arterial wall lipids, calcium, phospholipids, and aortic plaque area. In the prescription, it serves as the ministerial drug to promote blood circulation, and at the same time assists the monarch drug Astragalus membranaceus to enhance the effects of promoting diuresis, promoting blood circulation, and relieving pain, enhancing the relief effect on chest pain and palpitations symptoms. The assistant and guiding drugs are Atractylodes macrocephala Koidz. stir-fried with bran, Polygonatum sibiricum Delar. ex Redoute processed with wine, and Rhizoma Curcumae Aromaticae Vinegar-processed. Among them, Atractylodes macrocephala Koidz. stir-fried with bran dries dampness and promotes diuresis to assist the effects of the monarch and ministerial drugs; Polygonatum sibiricum Delar. ex Redoute processed with wine supplements qi and tonifies deficiency, especially good at nourishing yin, and can assist in controlling the monarch drug from generating dryness due to qi supplementation. Both drugs can enhance the effect of the monarch drug in treating shortness of breath and fatigue in patients with calcific aortic valve disease. And Curcuma zedoaria (Christm.) Rosc. is "the most important drug for treating various qi stagnations and accumulations" (Illustrated Classic of Materia Medica). It can promote blood circulation by breaking blood stasis and eliminating stagnation, assisting the monarch drug, and enhancing the therapeutic effect of the prescription on chest pain and palpitations. Thus, the compatibility of all the drugs in the whole prescription achieves the effects of supplementing qi and promoting blood circulation, strengthening the healthy qi and eliminating masses. The whole prescription targets the pathogenesis of "deficiency of healthy qi, mutual binding of phlegm and stasis, and accumulation of masses" in patients with calcific aortic valve disease, and can achieve the effects of "supplementing qi and promoting blood circulation, strengthening the healthy qi and eliminating masses".

[0056] The following further describes the present invention with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the scope of the present invention.

[0057] For those not specifying specific technologies or conditions in the embodiments, they shall be carried out according to the technologies or conditions described in the literature in this field or according to the product instructions. For reagents or instruments not indicating the manufacturer, they are all conventional products that can be obtained through regular channels.

[0058] The experimental methods in the following examples are all conventional methods unless otherwise specified. The test materials used in the following examples are all commercially available products unless otherwise specified. Detailed implementation manners Example 1

[0060] 20 g of raw Astragalus membranaceus, 12 g of red peony root, 15 g of stir-fried Atractylodes macrocephala, 8 g of wine-processed Polygonatum sibiricum, 8 g of vinegar-processed Rhizoma Curcumae

[0061] Total amount: 63 g

[0062] The raw herbs of raw Astragalus membranaceus, red peony root, stir-fried Atractylodes macrocephala, wine-processed Polygonatum sibiricum, and vinegar-processed Rhizoma Curcumae were pulverized into fine powder, and 500 mL of distilled water was added for decoction. After the first decoction for 1.5 hours, the first filtrate was obtained by filtration. Another 500 mL of distilled water was added to the residue for decoction, and after decocting for 1.5 hours, the second filtrate was obtained by filtration. The first filtrate and the second filtrate were combined and concentrated under reduced pressure to 50 mL.

[0063] Example 2:

[0064] 30 g of raw Astragalus membranaceus, 20 g of red peony root, 20 g of stir-fried Atractylodes macrocephala, 10 g of wine-processed Polygonatum sibiricum, 10 g of vinegar-processed Rhizoma Curcumae

[0065] Total amount: 90 g

[0066] Control group:

[0067] 40 g of raw Astragalus membranaceus, 20 g of red peony root

[0068] Total amount: 60 g

[0069] The preparation methods of Example 2 and the control group are the same as that of Example 1.

[0070] Western medicine group:

[0071] 20 mg of atorvastatin calcium tablets (the main ingredient is atorvastatin calcium, purchased from Pfizer Pharmaceuticals Limited, production batch number: 8141439)

[0072] Effect example:

[0073] 1. Experimental animals

[0074] In this study, male C57BL / 6J and ApoE at 6 - 8 weeks old were selected. - / -Mice, weighing 18 - 22 g, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. [License number: SCXK (Beijing) 2021 - 0006] and were housed in the animal facility of Xiyuan Hospital, China Academy of Chinese Medical Sciences [License number: SYXK (Beijing) 2023 - 0053]. The breeding temperature was maintained at 22°C ± 2°C, the humidity was 40% - 60%, and there was a 12 - hour day - night cycle. This experiment has been approved by the Animal Care and Use Committee of Xiyuan Hospital, China Academy of Chinese Medical Sciences, and was carried out strictly in accordance with the regulations of the ethics of laboratory animals and the Animal Management Association of Xiyuan Hospital, China Academy of Chinese Medical Sciences (Ethical review number: 2022XLC047 - 2).

[0075] 2. Construction of a mouse model of calcific aortic valve disease

[0076] ApoE - deficient mice were continuously fed with D12108C high - fat diet (containing 40% fat and 1.25% total cholesterol) for 16 weeks. - / -

[0077] 3. Grouping and intervention of experimental animals

[0078] After 7 days of adaptive feeding in a SPF - level mouse animal laboratory (barrier environment), ApoE - deficient mice were randomly divided into 5 groups (n = 6 in each group): model group, Example 1 group, Example 2 group, control group, and western medicine group. The 5 groups of mice were fed with high - fat diet for 12 weeks. At the end of the 12th week, 3 mice were randomly selected for ultrasonic detection to evaluate the calcification of the aortic valve. After confirming the successful establishment of the model, the mice were continued to be fed with high - fat diet, and the mice in Example 1 - 2 groups, control group, and western medicine group were given intragastric administration. - / - n

[0079] Example 1 group: The doses of Astragalus membranaceus, Paeonia lactiflora, Atractylodes macrocephala, Polygonatum sibiricum, and Rhizoma Curcumae were converted according to the human - mouse conversion coefficient to obtain the total drug amount required for each group of mice (n = 6) in 28 days. On this basis, an additional 20% of the loss amount of the drug was added to calculate the final drug amount. Assuming the human body weight is 70 kg and the mouse body weight is 0.03 kg, with a conversion coefficient of 12.33, and 6 mice in each group, a total of 28 days of intragastric administration was required for conversion. It was calculated that Example 1 required a total of 21.31 g of Astragalus membranaceus, 12.78 g of Paeonia lactiflora, 15.98 g of Atractylodes macrocephala, 8.52 g of Polygonatum sibiricum, and 8.52 g of Rhizoma Curcumae. These were decocted with water and concentrated to 60.48 mL (the intragastric administration volume for each mouse per day was 0.3 mL, 6 mice in each group, and a total of 28 days of intragastric administration, so the total volume of the concentrated solution was 60.48 mL). n

[0080] The calculation method of the drug dosage for the mice in Example 2 group and control group and the volume of the solution after concentration by decoction were the same as those in Example 1 group.

[0081] ​​​​Western medicine group: After grinding the drug, according to the same human-mouse body weight conversion rule, calculating with a 20% loss, a total of 21.3 g of atorvastatin calcium tablets powder was weighed. The powder was dissolved in 60.48 mL of normal saline and ultrasonicated to obtain a solution for standby.

[0082] Normal control group and model control group mice were given intragastric administration of the same volume (0.3 mL) of normal saline daily for 4 consecutive weeks. Among them, C57BL / 6J mice ( n =6) were fed with ordinary maintenance feed, and ApoE - / - mice continued to be fed with high-fat feed. The specific feeding and administration regimens are shown in Table 1.

[0083] Table 1: Administration regimens used in this experiment

[0084]

[0085] 4. Detection indexes and statistical analysis methods

[0086] 4.1 Small animal ultrasound detection

[0087] The ultrasound images at the aortic valve of mice were detected by the Vevo 2100 high-resolution visual ultrasound imaging system. Mice were anesthetized with 2% isoflurane and maintained with 1.5% isoflurane, and the mice were fixed in a supine position on an operating table with appropriate temperature. When performing ultrasound evaluation, the heart rate of the mice was maintained at 400 - 450 beats per minute.

[0088] The ultrasound probe (30 MHz) was longitudinally applied under the xiphoid process of the sternum to identify the heart, and then slid upward to find the position of the aortic valve. Subsequently, under the guidance of the two-dimensional long-axis image, transthoracic B-mode ultrasound was obtained, and all measurements lasted for at least 5 consecutive cardiac cycles. The peak flow velocity before and after the valve was obtained from the B-mode ultrasound image, and the transvalvular pressure difference was calculated. The result of each measurement was at least the average value of three consecutive cardiac cycles.

[0089] 4.2 Sample collection

[0090] After 4 weeks of administration, the mice in each group were fasted but not water-restricted for 12 h, and then ultrasound detection was performed. Before the mice recovered from anesthesia after the detection, blood was collected by eye enucleation. The whole blood sample was left standing at 4°C for 1 - 2 h, and then centrifuged at a speed of 2500 rpm for 20 min in a pre-cooled 4°C centrifuge. The supernatant after centrifugation was transferred to a 1.5 mL EP tube and stored in a -20°C refrigerator for later inspection. Then, the abdominal cavity and thoracic cavity of the mice were opened in turn. After removing the lungs, trachea, etc., the heart was exposed, and the heart was perfused from the apex with 5 mL of pre-cooled normal saline to wash the blood remaining in the heart. After washing, the heart together with a part of the aortic arch was removed, and the sample was fixed in 4% paraformaldehyde fixative for later pathological and immunohistochemical staining.

[0091] 4.3 Detection of Aortic Valve Calcification by Calcium Salt Staining

[0092] After tissue dehydration and embedding, after dewaxing, irradiate with strong light in Von Kossa silver solution for 40 min, wash with ultrapure water for 1 min, then treat with hypo solution for 2 min, and counterstain the tissue with HE staining solution. Then dehydrate, clear, and mount with neutral balsam for the Von Kossa staining process. For alizarin red (AR) staining, after dewaxing to 95% ethanol, stand the slides upright and air-dry them. Stain the air-dried slides with alizarin red S for 2 min, quickly rinse with ultrapure water, then counterstain with fast green staining solution for 30 s. After washing with ultrapure water 3 times, dehydrate, clear, and mount with neutral balsam.

[0093] 4.4 Detection of Aortic Valve Calcification Markers in Mice by Immunohistochemistry

[0094] After tissue dehydration and embedding, after dewaxing, perform heat repair with EDTA antigen retrieval solution. After inactivating endogenous peroxidase, dilute the primary antibodies of RUNX2 and BMP2 with PBS solution at a ratio of 1:200, add the primary antibodies, and incubate overnight at 4°C. After rinsing off the primary antibodies with PBS, add the secondary antibody and incubate at room temperature for 1 h. After rinsing with PBS, perform DAB color development and nuclear counterstaining. After dehydration, clearing, and mounting, take pictures under an upright microscope, and use Image J to calculate the positive staining rate.

[0095] 4.5 Statistical Analysis

[0096] Use SPSS 24.0 software to statistically analyze the cardiac ultrasound and protein data. First, determine whether it conforms to a normal distribution. For data that conforms to a normal distribution, perform one-way ANOVA, and conduct a homogeneity of variance test. When the variances are homogeneous, use the Bonferroni test, and when the variances are not completely homogeneous, use the Dunnett test for non-parametric testing. When P <0.05, it is considered that there are significant differences in the experimental results.

[0097] 5. Experimental Results

[0098] (1) Detection of Ultrasonic Results of Mouse Aortic Valve

[0099] Use a small animal ultrasonic detector to detect the peak velocity and transvalvular pressure difference of the aortic valve in each group of mice. The severity of CAVD disease increases with the increase in the values of peak velocity and transvalvular pressure difference. The experimental results (see Table 2) show that the peak velocity and transvalvular pressure difference of the aortic valve in the model control group of mice are higher than those in the blank control group ( P<0.05), indicating that aortic valve stenosis did occur in the mice of the model control group. After drug intervention in the Example 1 group, Example 2 group, and comparative example group, the peak velocity could be significantly reduced ( P< 0.05), among which, after drug intervention in the Example 2 group, the improvement of peak velocity was the most significant. For the transvalvular pressure gradient, the drugs in the Example 1 group, Example 2 group, and comparative example group could all significantly reduce the transvalvular pressure gradient of CAVD mice ( P< 0.01). Among them, after drug intervention in the Example 2 group, the improvement of transvalvular pressure gradient was the most obvious, followed by the Example 1 group, and then the comparative example group. After intervention with western medicine, the peak velocity and transvalvular pressure gradient were not significantly reduced. The results showed that after intervention in the Example 2 group, the degree of aortic valve stenosis improved the most significantly, followed by the drugs in the Example 1 group and comparative example group in improving the degree of aortic valve stenosis, and the effect of the western medicine group was not good.

[0100] Table 2: Echocardiographic assessment of aortic valve ( n =6)

[0101]

[0102] Note: : Compared with the blank group, P <0.01; : Compared with the blank group, P <0.05; △△ : Compared with the model group, P <0.01; △ : Compared with the model group, P <0.05.

[0103] (2) Pathological staining results of aortic valve calcification in mice

[0104] In addition to the pathological manifestations of aortic valve stenosis, CAVD will also present aortic valve calcification. Von Kossa staining and alizarin red staining are pathological staining methods for detecting the degree of aortic valve calcification. After staining, the calcified parts in the aortic valve will appear brown. With the increase of calcification degree, the brown color will deepen to a certain extent; with the increase of calcification area, the brown staining area will increase accordingly. The pathological changes of the aortic valve in each group of mice were detected by Von Kossa staining and alizarin red staining, as Figure 1 - Figure 2 shown.

[0105] The pathological staining results (see Table 3) indicated that the areas of Von Kossa staining and alizarin red staining of the aortic valve in the mice of the model control group were significantly increased compared with those of the blank control group ( P <0.01), suggesting a higher degree of aortic valve calcification in the mice of the model control group. After intervention in the Example 1 group, the areas of Von Kossa staining and alizarin red staining could be significantly reduced ( P< 0.05, P < 0.01). After the intervention in the Example 2 group, the areas of Von Kossa staining and alizarin red staining can be significantly reduced ( P < 0.01), and the effect is better than that of the Example 1 group. After the drug intervention in the control group and the western medicine group, the areas of Von Kossa and alizarin red staining in CAVD mice cannot be improved, that is, the aortic valve calcification cannot be significantly improved. Based on the above results, after the intervention in the Example 2 group, the reduction of the aortic valve calcification area is the most obvious, the effect of the Example 1 group is the second, and the drugs in the control group and the western medicine group have no significant effect on improving aortic valve calcification.

[0106] Table 3: Pathological staining results of aortic valve calcification degree

[0107]

[0108] Note: : Compared with the blank group, P < 0.01; : Compared with the blank group, P < 0.05; △△ : Compared with the model group, P < 0.01; △ : Compared with the model group, P < 0.05.

[0109] (3) Immunohistochemical detection of the levels of aortic valve calcification markers in mice

[0110] RUNX2 and BMP2 are markers of calcification. As the degree of calcification increases, the immunohistochemical staining area and degree increase. The relative expression levels of aortic valve calcification markers in each group of mice were detected by immunohistochemistry (see Table 4), and the representative figures are as Figure 3 - Figure 4 shown. The staining areas (brown staining parts) of the aortic valve calcification markers RUNX2 and BMP2 in the model control group were significantly increased compared with the blank control group ( P < 0.01), indicating that obvious calcification occurred in the aortic valve of the model control group mice. After the intervention in the Example 1 and Example 2 groups, the staining areas of RUNX2 and BMP2 can be significantly reduced, that is, the expression levels of calcification markers are reduced ( P < 0.01). After the drug intervention in the control group and the western medicine group, the staining level of RUNX2 can be reduced ( P < 0.05), while there is no statistical difference in the expression level of BMP2 compared with the model control group. It can be seen that the Example 2 group has the most obvious reduction in the expression level of aortic valve calcification markers and the highest improvement in the degree of aortic valve calcification. The Example 1 group is the second, and the improvement degree of the drugs in the control group and the western medicine group on CAVD needs to be further observed.

[0111] Table 4: Immunohistochemical staining results of aortic valve calcification markers

[0112]

[0113] Note: : Compared with the blank group, P <0.01; : Compared with the blank group, P <0.05; △△ : Compared with the model group, P <0.01; △ : Compared with the model group, P <0.05.

[0114] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalent technologies, the present invention is also intended to include these changes and modifications.

Claims

1. A Chinese medicine composition for treating calcific aortic valve disease, characterized in that: The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 10-50 parts of raw astragalus, 5-40 parts of red peony root, 5-40 parts of stir-fried atractylodes, 1-20 parts of polygonatum odoratum, and 1-20 parts of vinegar-curcuma.

2. The Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared from the following raw medicines in parts by weight: 15-40 parts of raw astragalus, 10-30 parts of red peony root, 10-30 parts of stir-fried atractylodes, 5-15 parts of wine polygonatum, and 5-15 parts of vinegar curcuma.

3. The Chinese medicine composition according to claim 2, characterized in that: The traditional Chinese medicine composition is prepared from the following raw medicines in parts by weight: 20-30 parts of raw astragalus, 12-20 parts of red peony root, 15-20 parts of stir-fried atractylodes, 8-10 parts of wine polygonatum, and 8-10 parts of vinegar curcuma.

4. The Chinese medicine composition according to claim 1, characterized in that: The traditional Chinese medicine composition has the effects of invigorating qi and activating blood circulation, strengthening the body and eliminating symptoms.

5. The method for preparing the Chinese medicine composition according to any one of claims 1 to 4, characterized in that: The preparation method of the traditional Chinese medicine composition comprises the following steps: weighing each raw material medicine according to the weight proportions described in any one of claims 1 to 3, decocting, and concentrating.

6. The preparation method according to claim 5, characterized in that: The decoction is performed twice. After the first decoction for 1-3 hours, a primary filtrate is obtained by filtering. The filter residue is then decocted with water for 1-3 hours, and a secondary filtrate is obtained by filtering. The primary filtrate and the secondary filtrate are combined.

7. The preparation method according to claim 6, characterized in that: The water used for the first decoction and the second decoction is distilled water; and the concentration is reduced-pressure concentration.

8. Use of the Chinese medicine composition according to any one of claims 1 to 4 or the Chinese medicine composition prepared by the preparation method according to any one of claims 5 to 7 in the preparation of a medicament for treating calcific aortic valve disease.

9. The use according to claim 8, characterized in that: The medicine is in the form of decoction pieces, decoctions, powders, pastes, granules, tablets, oral liquids or capsules.

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