阅读说明：本技术 一种用于缺血性心肌病的中药组合物及其应用 (Traditional Chinese medicine composition for ischemic cardiomyopathy and application thereof ) 是由 王�忠 王永炎 陈林 于 2021-08-25 设计创作，主要内容包括：本发明涉及一种用于缺血性心肌病的中药组合物,包括葛根1-25份、石韦1-25份、银杏叶1-20份、紫苏梗1-15份、木贼1-15份、虎杖1-15份、鸡骨草1-15份。所述中药组合物还可以包括葛根1-25份。本发明提供的中药组合物对心衰斑马鱼的心脏功能改善作用和对心肌损伤斑马鱼的心肌保护作用。(The invention relates to a traditional Chinese medicine composition for ischemic cardiomyopathy, which comprises 1-25 parts of kudzuvine root, 1-25 parts of pyrrosia lingua, 1-20 parts of ginkgo leaf, 1-15 parts of perilla stem, 1-15 parts of equisetum hiemale, 1-15 parts of giant knotweed rhizome and 1-15 parts of abrus cantoniensis hance. The traditional Chinese medicine composition can also comprise 1-25 parts of kudzu root. The traditional Chinese medicine composition provided by the invention has the effect of improving the heart function of heart failure zebra fish and has the effect of protecting the myocardium of the zebra fish with myocardial injury.)

1. A traditional Chinese medicine composition for ischemic cardiomyopathy comprises the following medicinal materials in parts by weight:

1-25 parts of pyrrosia lingua, 1-20 parts of ginkgo leaf, 1-15 parts of perilla stem, 1-15 parts of equisetum, 1-15 parts of giant knotweed and 1-15 parts of abrus cantoniensis hance;

preferably, the traditional Chinese medicine composition comprises the following medicinal materials in parts by weight:

10-20 parts of pyrrosia lingua, 10-15 parts of ginkgo leaf, 5-10 parts of perilla stem, 5-10 parts of equisetum, 5-10 parts of giant knotweed and 5-10 parts of abrus cantoniensis hance.

2. The traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition comprises the following medicinal materials in parts by weight:

18 parts of pyrrosia lingua, 15 parts of ginkgo leaves, 9 parts of perilla stems, 9 parts of equisetum, 9 parts of polygonum cuspidatum and 9 parts of abrus cantoniensis hance.

3. The traditional Chinese medicine composition according to claim 1 or 2, wherein the traditional Chinese medicine composition is composed of pyrrosia lingua, ginkgo leaf, perilla stem, equisetum, polygonum cuspidatum and abrus cantoniensis hance, and the weight parts of the medicinal materials are defined in claim 1 or 2.

4. A traditional Chinese medicine composition for ischemic cardiomyopathy comprises the following medicinal materials in parts by weight:

1-25 parts of kudzu root, 1-25 parts of pyrrosia lingua, 1-20 parts of ginkgo leaf, 1-15 parts of perilla stem, 1-15 parts of equisetum, 1-15 parts of giant knotweed and 1-15 parts of abrus cantoniensis hance;

preferably, the traditional Chinese medicine composition comprises the following medicinal materials in parts by weight:

10-20 parts of kudzu root, 10-20 parts of pyrrosia lingua, 10-15 parts of ginkgo leaf, 5-10 parts of perilla stem, 5-10 parts of equisetum, 5-10 parts of giant knotweed and 5-10 parts of abrus herb.

5. The traditional Chinese medicine composition according to claim 4, which is characterized by comprising the following medicinal materials in parts by weight:

18 parts of kudzuvine root, 18 parts of pyrrosia lingua, 15 parts of ginkgo leaf, 9 parts of perilla stem, 9 parts of equisetum, 9 parts of giant knotweed rhizome and 9 parts of abrus cantoniensis hance.

6. The Chinese medicinal composition according to claim 4 or 5, wherein the Chinese medicinal composition is prepared from kudzu root, pyrrosia lingua, ginkgo leaf, perilla stem, equisetum, giant knotweed rhizome and abrus herb, and the weight parts of the medicinal materials are defined in claim 4 or 5.

7. A medicament comprising the Chinese medicinal composition of any one of claims 1 to 6, with or without pharmaceutically acceptable adjuvants;

preferably, the medicament is any one clinically acceptable preparation;

more preferably, the formulation is an oral formulation;

further preferably, the oral preparation is selected from one or more of decoction, powder, capsules, tablets, honey pills, water pills, concentrated pills, paste pills, wax pills, granules, oral liquid or dropping pills.

8. The preparation method of the medicament of claim 7, which comprises the steps of preparing the medicinal materials according to the parts by weight, adding or not adding pharmaceutically acceptable auxiliary materials according to the conventional method in the field, and preparing into clinically acceptable preparations.

9. Use of the Chinese medicinal composition of any one of claims 1 to 6, the medicament of claim 7 or the medicament prepared by the preparation method of claim 8 in the preparation of a medicament for treating ischemic cardiomyopathy;

preferably, the ischemic cardiomyopathy comprises congestive ischemic cardiomyopathy and restrictive ischemic cardiomyopathy.

Technical Field

The invention belongs to the technical field of traditional Chinese medicines and medicine and pharmacology, and particularly relates to a traditional Chinese medicine composition for treating ischemic cardiomyopathy and application thereof.

Background

Ischemic Cardiomyopathy (ICM) belongs to a special type or an advanced stage of coronary heart disease, and refers to a clinical syndrome that coronary atherosclerosis is formed due to various reasons, myocardial long-term chronic ischemia and hypoxia occur, myocardial necrosis occurs, diffuse myocardial fibrosis is caused, myocardial damage is caused, the heart is enlarged, the functions of systole and diastole are damaged, and arrhythmia and heart failure are finally caused. The clinical classification is full blood type ischemic cardiomyopathy and restrictive ischemic cardiomyopathy according to different manifestations of patients. Epidemiological data indicate that the prevalence of ICMs is 2600 million worldwide, with a prevalence of 195.3/10 million for men and 115.0/10 million for women. Ischemic cardiomyopathy has become one of the leading causes of death in cardiovascular disease and must therefore be highly appreciated by medical workers and patients.

The pathogenesis of ischemic cardiomyopathy is not completely clarified so far, but researches show that a plurality of factors such as inflammatory reaction, oxidative stress, mitochondrial dysfunction, calcium overload, fibrosis, endoplasmic reticulum stress, autophagy and apoptosis, proteomics, signal pathways and the like participate in the pathogenesis, and the factors interact and influence with each other to jointly promote the occurrence and development of ICM.

The prognosis of ischemic cardiomyopathy is poor, and the internal treatment mainly aims at improving the cardiac function and relieving symptoms. Although modern medicine adopts various treatment means and the treatment of ischemic cardiomyopathy has great progress, the side effects of bleeding, liver function damage and the like and economic pressure caused by drug treatment bring great burden to patients. Surgical treatment of ICMs is more limited.

Various traditional Chinese medicine compositions for treating ischemic cardiomyopathy appear in the prior art. For example, chinese invention patent publication No. CN101461843B (publication No. 2011/6/8) discloses a pharmaceutical composition for ischemic diseases (including ischemic cardiomyopathy), which comprises the following raw materials in parts by weight: 5-15 parts of astragalus root, 2-8 parts of salvia miltiorrhiza, 1-5 parts of ligusticum wallichii, 1-5 parts of safflower and 1-3 parts of earthworm. The Chinese invention patent publication No. CN101357125B (publication No. 2012/2/8) provides the use of leonurine in preparing medicine for treating heart failure. Chinese patent publication No. CN106727873B (6/19/2020), discloses an oral dosage form pharmaceutical composition for preventing and/or treating ischemic cardiomyopathy, which contains hedysarum polybotrys or hedysarum polybotrys polysaccharides as a single active ingredient.

However, the clinical treatment requirements for ischemic cardiomyopathy are not met, and the requirements for drugs with good curative effects and few side effects are still very urgent.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention provides a traditional Chinese medicine composition for treating ischemic cardiomyopathy. The traditional Chinese medicine composition has novel formulation thought. Preliminary experiments show that the traditional Chinese medicine composition has a relatively obvious myocardial protection effect on the zebra fish with myocardial injury and an improvement effect on the cardiac function of the zebra fish with heart failure, and the application of the traditional Chinese medicine composition in treating ischemic cardiomyopathy is prompted.

Therefore, the invention adopts the following technical scheme:

a traditional Chinese medicine composition for ischemic cardiomyopathy comprises the following components in parts by weight:

1-25 parts of pyrrosia lingua, 1-20 parts of ginkgo leaf, 1-15 parts of perilla stem, 1-15 parts of equisetum, 1-15 parts of giant knotweed and 1-15 parts of abrus cantoniensis hance.

Preferably, the traditional Chinese medicine composition for ischemic cardiomyopathy comprises the following components in parts by weight:

10-20 parts of pyrrosia lingua, 10-15 parts of ginkgo leaf, 5-10 parts of perilla stem, 5-10 parts of equisetum, 5-10 parts of giant knotweed and 5-10 parts of abrus cantoniensis hance.

More preferably, the traditional Chinese medicine composition for ischemic cardiomyopathy comprises the following components in parts by weight:

18 parts of pyrrosia lingua, 15 parts of ginkgo leaves, 9 parts of perilla stems, 9 parts of equisetum, 9 parts of polygonum cuspidatum and 9 parts of abrus cantoniensis hance.

As a preferred embodiment, the traditional Chinese medicine composition for ischemic cardiomyopathy of the invention comprises pyrrosia lingua, ginkgo leaves, perilla stems, equisetum, giant knotweed rhizome and abrus cantoniensis hance, and the weight parts of the raw materials are defined as above.

The invention also provides a traditional Chinese medicine composition for treating ischemic cardiomyopathy, which comprises the following components in parts by weight:

1-25 parts of kudzu root, 1-25 parts of pyrrosia lingua, 1-20 parts of ginkgo leaf, 1-15 parts of perilla stem, 1-15 parts of equisetum, 1-15 parts of giant knotweed and 1-15 parts of abrus cantoniensis hance.

Preferably, the traditional Chinese medicine composition for ischemic cardiomyopathy comprises the following components in parts by weight:

10-20 parts of kudzu root, 10-20 parts of pyrrosia lingua, 10-15 parts of ginkgo leaf, 5-10 parts of perilla stem, 5-10 parts of equisetum, 5-10 parts of giant knotweed and 5-10 parts of abrus herb.

More preferably, the traditional Chinese medicine composition for ischemic cardiomyopathy comprises the following components in parts by weight:

18 parts of kudzuvine root, 18 parts of pyrrosia lingua, 15 parts of ginkgo leaf, 9 parts of perilla stem, 9 parts of equisetum, 9 parts of giant knotweed rhizome and 9 parts of abrus cantoniensis hance.

As a preferred embodiment, the traditional Chinese medicine composition for ischemic cardiomyopathy of the invention comprises kudzu root, pyrrosia lingua, ginkgo leaf, perilla stem, equisetum, giant knotweed rhizome and abrus cantoniensis hance, and the weight parts of the raw materials are defined as above.

The invention also aims to provide a medicament, which comprises the traditional Chinese medicine composition for treating ischemic cardiomyopathy and optionally pharmaceutically acceptable auxiliary materials.

Preferably, the medicament is any one clinically acceptable preparation.

More preferably, the medicament is an oral formulation.

Further preferably, the oral preparation is selected from one or more of decoction, powder, capsule, tablet, honey pill, water-honey pill, concentrated pill, paste pill, wax pill, granule, oral liquid and drop pill.

The invention also aims to provide a preparation method of the medicine, which comprises the steps of preparing the raw materials according to the parts by weight, extracting or not extracting according to a conventional method in the field, and adding or not adding pharmaceutically acceptable auxiliary materials to prepare a clinically acceptable preparation.

In addition, the invention also provides application of the traditional Chinese medicine composition, the medicine or the medicine prepared by the preparation method in preparing a medicine for treating ischemic cardiomyopathy.

The ischemic cardiomyopathy comprises congestive ischemic cardiomyopathy and restrictive ischemic cardiomyopathy. The pharmaceutically acceptable excipients of the present invention include, but are not limited to, (1) diluents, examples

Such as starch, sugar powder, dextrin, lactose, pregelatinized starch, microcrystalline fiber, inorganic calcium salt (such as calcium sulfate, calcium hydrogen phosphate, medicinal calcium carbonate, etc.), mannitol, etc., vegetable oil, polyethylene glycol, etc.; (2) binders such as distilled water, ethanol, starch slurry, sodium carboxymethylcellulose, hydroxypropyl cellulose, methyl and ethyl cellulose, hypromellose, and the like; (3) disintegrants such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, croscarmellose sodium and the like; (4) lubricants such as magnesium stearate, aerosil, talc, hydrogenated vegetable oils, polyethylene glycols, magnesium lauryl sulfate, and the like; (5) solvents such as water, ethanol solution, etc.

In the present specification, "parts by weight" or "parts" of each component means a quantitative proportioning relationship between the components, not an actual mass unit. The unit weight or each part may be of any mass, as appropriate, e.g. 1 part by weight or 1 part may be 1g, 500g or 1kg, and may even be 15g, 30g etc.

The traditional Chinese medicine composition for ischemic cardiomyopathy provided by the invention comprises the following components in parts by weight:

ku Ge is sweet, pungent and cool in nature, entering spleen and stomach meridians and dispersing and ascending, so it is also wind-herbs in nature. In the statement of materia medica, Kudzuvine root, its cool powder is good at all yang meridians, but yangming is the most important one; it is good at relieving exterior syndrome and inducing sweat because it is light in smell. In this recipe, the actions of dispersing the pathogenic factors from the exterior and relieving the pathogenic factors from the muscles are well-known. Ginkgo leaves, which are sweet, bitter, astringent and even in nature, enter heart and lung meridians, are recorded in the Chinese pharmacopoeia and are good at astringing lung, relieving asthma, promoting blood circulation to remove blood stasis and relieving pain. If the two medicines are used together, the effects of expelling pathogenic factors, relieving muscles and promoting qi and blood circulation are achieved.

Ji Guo is indicated for its actions of clearing heat and removing toxicity, soothing liver and alleviating pain.

Horsetail, herba Equiseti hiemalis, sweet in nature, bitter in taste, neutral in nature, enters lung and liver meridians, and has the effects of inducing sweat, expelling pathogenic factors from muscles and treating diseases such as transverse invasion of liver and gallbladder pathogens according to records in materia medica Zhengyi, so that the horsetail is combined with kudzu root to strengthen the efficacy of expelling pathogenic factors and expelling muscles, is combined with Abrus cantoniensis Hance to relieve depression of liver meridians, and is combined with ginkgo leaf and giant knotweed to strengthen the efficacy of promoting qi and activating blood circulation.

Shi Wei enters lung and bladder meridians due to its bitter and slightly cold nature. Pyrrosia lingua regulates qi movement of the whole body by regulating the function of the lung, so that blood can normally run.

Polygonum cuspidatum is bitter and slightly cold in nature, enters liver, gallbladder and lung channels, and has the effect of breaking and retaining blood stagnation, and is combined with ginkgo leaves to play the role of promoting blood circulation and removing blood stasis.

Perilla stem, pungent and sweet in flavor, slightly warm. It enters lung, spleen and stomach meridians and has the actions of regulating qi and relieving depression. It is combined with Equisetum hiemale and abrus herb to achieve the effect of soothing liver and promoting qi; it is combined with ginkgo leaf and giant knotweed rhizome to promote blood circulation. Moreover, the perilla stem is slightly warm, which restricts the cooling property of other herbs and prevents the recipe from being too cold and cold to injure the stomach.

The medicines are combined to play the roles of expelling pathogenic factors and relieving muscles, promoting qi and activating blood, cutting the pathogenesis of ischemic cardiomyopathy and treating both principal and secondary aspects of diseases. The traditional Chinese medicine composition has a function improvement effect on the heart of the heart failure zebra fish and a protection effect on the cardiac muscle of the zebra fish with cardiac muscle injury, so that the traditional Chinese medicine composition can protect the cardiac muscle and improve the cardiac function, and therefore ischemic cardiomyopathy is treated.

Detailed Description

The invention is illustrated below with reference to specific examples. It will be understood by those skilled in the art that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention in any way.

The experimental procedures in the following examples are conventional unless otherwise specified. The raw materials and reagents used in the following examples are all commercially available products unless otherwise specified.

The medicinal materials used in the following examples are all commercially available decoction pieces.

Example 1A Chinese medicinal composition for ischemic cardiomyopathy

The traditional Chinese medicine composition of the embodiment comprises the following raw materials (1 part:

18 parts of pyrrosia lingua, 15 parts of ginkgo leaves, 9 parts of perilla stems, 9 parts of equisetum, 9 parts of polygonum cuspidatum and 9 parts of abrus cantoniensis hance.

Example 2A Chinese medicinal composition for ischemic cardiomyopathy

The traditional Chinese medicine composition of the embodiment comprises the following raw materials (1 part:

16 parts of pyrrosia lingua, 10 parts of ginkgo leaves, 6 parts of perilla stems, 6 parts of equisetum, 6 parts of polygonum cuspidatum and 6 parts of abrus cantoniensis hance.

Example 3A Chinese medicinal composition for ischemic cardiomyopathy

The raw materials of the traditional Chinese medicine composition of the embodiment are (1 part to 1 kg):

25 parts of pyrrosia lingua, 20 parts of ginkgo leaves, 15 parts of perilla stems, 15 parts of equisetum, 15 parts of polygonum cuspidatum and 15 parts of abrus cantoniensis hance.

Example 4A Chinese medicinal composition for ischemic cardiomyopathy

The raw materials of the traditional Chinese medicine composition of the embodiment are (1 part to 1 kg):

10 parts of pyrrosia lingua, 10 parts of ginkgo leaf, 5 parts of perilla stem, 5 parts of horsetail, 5 parts of giant knotweed and 5 parts of abrus herb.

Example 5A Chinese medicinal composition for treating ischemic cardiomyopathy

The traditional Chinese medicine composition of the embodiment comprises the following raw materials (1 part:

20 parts of pyrrosia lingua, 15 parts of ginkgo leaves, 10 parts of perilla stems, 10 parts of equisetum, 10 parts of polygonum cuspidatum and 10 parts of abrus cantoniensis hance.

Example 6A Chinese medicinal composition for ischemic cardiomyopathy

The traditional Chinese medicine composition of the embodiment comprises the following raw materials (1 part:

18 parts of kudzuvine root, 18 parts of pyrrosia lingua, 15 parts of ginkgo leaf, 9 parts of perilla stem, 9 parts of equisetum, 9 parts of giant knotweed rhizome and 9 parts of abrus cantoniensis hance.

Example 7A Chinese medicinal composition for treating ischemic cardiomyopathy

The traditional Chinese medicine composition of the embodiment comprises the following raw materials (1 part ═ 500 g):

16 parts of kudzuvine root, 16 parts of pyrrosia lingua, 10 parts of ginkgo leaf, 6 parts of perilla stem, 6 parts of equisetum, 6 parts of giant knotweed rhizome and 6 parts of abrus cantoniensis hance.

Example 8A Chinese medicinal composition for treating ischemic cardiomyopathy

The raw materials of the traditional Chinese medicine composition of the embodiment are (1 part to 1 kg):

25 parts of kudzu root, 25 parts of pyrrosia lingua, 20 parts of ginkgo leaf, 15 parts of perilla stem, 15 parts of equisetum, 15 parts of giant knotweed rhizome and 15 parts of abrus cantoniensis hance.

Example 9A Chinese medicinal composition for treating ischemic cardiomyopathy

The traditional Chinese medicine composition of the embodiment comprises the following raw materials (1 part:

10 parts of kudzu root, 10 parts of pyrrosia lingua, 10 parts of ginkgo leaf, 5 parts of perilla stem, 5 parts of equisetum, 5 parts of giant knotweed rhizome and 5 parts of abrus cantoniensis hance.

Example 10A Chinese medicinal composition for ischemic cardiomyopathy

The traditional Chinese medicine composition of the embodiment comprises the following raw materials (1 part:

20 parts of kudzu root, 20 parts of pyrrosia lingua, 15 parts of ginkgo leaf, 10 parts of perilla stem, 10 parts of equisetum, 10 parts of giant knotweed rhizome and 10 parts of abrus cantoniensis hance.

EXAMPLE 11A capsule

The traditional Chinese medicine composition of the embodiment 1 is taken, mixed and ground into fine powder, the fine powder is sieved by a 100-mesh sieve and is filled into capsules, and 0.6g of crude drugs are taken per capsule to obtain the traditional Chinese medicine composition.

The usage method comprises the following steps: the preparation is taken three times a day, 2-4 granules each time, and is taken with warm water.

EXAMPLE 12A capsule

The traditional Chinese medicine composition in the embodiment 6 is taken, mixed and ground into fine powder, the fine powder is sieved by a 100-mesh sieve and is filled into capsules, and 0.6g of crude drugs are taken per capsule to obtain the traditional Chinese medicine composition.

The usage method comprises the following steps: the preparation is taken three times a day, 2-4 granules each time, and is taken with warm water.

Example 13A tablet

Mixing the Chinese medicinal composition of example 3, grinding into fine powder, sieving with 80-100 mesh sieve, adding dextrin, and directly pressing into tablet, 0.4g crude drug/tablet.

The usage method comprises the following steps: taken with warm water three times a day, 2-4 tablets each time.

EXAMPLE 14A tablet

Mixing the Chinese medicinal composition of example 7, grinding into fine powder, sieving with 80-100 mesh sieve, adding dextrin, and directly pressing into tablet, 0.4g crude drug/tablet.

The usage method comprises the following steps: taken with warm water three times a day, 2-4 tablets each time.

Example 15A Honey pill

Mixing the traditional Chinese medicine composition of the embodiment 4, grinding into fine powder, sieving with a 60-100 mesh sieve, adding appropriate refined honey, and preparing into honeyed pills with a dosage of 5g/10 pills according to a conventional method.

The usage method comprises the following steps: taken with warm water three times a day, 5g each time.

Example 16A Honey pill

Mixing the traditional Chinese medicine composition of the embodiment 8, grinding into fine powder, sieving with a 60-100 mesh sieve, adding appropriate refined honey, and preparing into honeyed pills with a dosage of 5g/10 pills according to a conventional method.

The usage method comprises the following steps: taken with warm water three times a day, 5g each time.

Embodiment 17A decoction

Weighing the following medicinal materials in parts by weight (1 g-1):

18g of pyrrosia lingua, 15g of ginkgo leaves, 9g of perilla stems, 9g of equisetum, 9g of giant knotweed and 9g of abrus herb.

Mixing the medicinal materials, adding 5-10 times of water by weight, heating to boil, decocting for 20-30 minutes, and filtering while heating; and adding 5-10 times of water into the decoction dregs, heating to boil, decocting for 20-30 minutes, filtering while heating, and combining the liquid medicines obtained in two times. The medicine is taken 2-3 times in 1 day.

Embodiment 18A decoction

Weighing the following medicinal materials in parts by weight (1 g-1):

18g of kudzuvine root, 18g of pyrrosia lingua, 15g of ginkgo leaf, 9g of perilla stem, 9g of equisetum, 9g of giant knotweed rhizome and 9g of abrus herb.

Mixing the medicinal materials, adding 5-10 times of water by weight, heating to boil, decocting for 20-30 minutes, and filtering while heating; and adding 5-10 times of water into the decoction dregs, heating to boil, decocting for 20-30 minutes, filtering while heating, and combining the liquid medicines obtained in two times. The medicine is taken 2-3 times in 1 day.

Test example 1 pharmacodynamic test

1. Material

1.1 Experimental drugs:

the contents of the capsule prepared in example 12, brown powder, were dried and stored in a cool cabinet as provided by the clinical basic research institute of the Chinese academy of science of traditional Chinese medicine. When in use, the capsule content is prepared into 100mg/mL mother solution by 10% DMSO, the final DMSO concentration in the working solution is 1%, and the capsule content is prepared at present.

The contents of the capsule prepared in example 11, brown powder, were dried and stored in a cool cabinet as provided by the clinical basic research institute of the Chinese academy of science of traditional Chinese medicine. When in use, the capsule content is prepared into 100mg/mL mother solution by 10% DMSO, the final DMSO concentration in the working solution is 1%, and the capsule content is prepared at present.

Isoproterenol hydrochloride, white powder, batch No. MZSVB-DI, purchased from TCI, Japan, stored in a cool cabinet protected from light. Preparing mother liquor with concentration of 200mg/mL with ultrapure water for use at present.

Digoxin, white powder, batch number L1303078, from Aladdin, was stored at 4 ℃. The concentration of the mother solution is 0.8mg/mL when 100% DMSO is prepared for clinical use, the concentration of DMSO in the final working solution is 1%, and the mother solution is stored at-20 ℃.

N-acetylcysteine (NAC for short), white powder with the batch number of A105420 and purchased from alatin, mother liquor with the concentration of 50mg/mL is prepared for standby by ultrapure water at the time of use, the DMSO concentration in the final working solution is 1 percent, and the working solution is prepared at the time of use.

1.2 Experimental animals

Wild type AB strain zebrafish, in a natural mated mating breeding mode. The age was 2 days after fertilization (2dpf) and was 780 tails, 30 tails per experimental group. The method is used for measuring the maximum tolerance concentration of the test article to the myocardial injury zebra fish.

Wild type AB strain zebrafish, in a natural mated mating breeding mode. The age was 2 days post fertilization (2dpf) for 225 tails, 15 tails per experimental group. Is used for evaluating the heart function improvement effect of the test sample on the coronary heart disease zebra fish.

Wild type AB strain zebrafish, in a natural mated mating breeding mode. The age was 2 days after fertilization (2dpf), for a total of 450 tails, 30 tails per experimental group. Is used for evaluating the myocardial protection effect of the test article on the myocardial-injured zebra fish.

The zebra fish are all raised in water for fish culture at 28 ℃ (water quality: 200mg of instant sea salt is added into per 1L of reverse osmosis water, the conductivity is 480-510 muS/cm, the pH is 6.9-7.2, and the hardness is 53.7-71.6 mg/L (CaCO)₃) The license number for experimental animals is as follows: SYXK (Zhe) 2012-0171. The feeding management meets the requirements of international AAALAC certification.

1.3 instruments and reagents

Dissecting microscopes (SZX7, OLYMPUS, Japan); a heartbeat blood flow analysis system (zebralab3.3(PB 2084C)); electric focusing continuous zoom fluorescence microscope (AZ100, Nikon, Japan); 6-well plates (Nest, China); 12-well plates (Nest, China); precision electronic balances (CP214, OHAUS, America); dimethyl sulfoxide (DMSO, Sigma, lot No. BCBT 0803); methylcellulose (Sigma, lot No. 079K 0054V); AO dye (Alorich, 494-38-2).

2. Method of producing a composite material

2.1 experiment 1: determining the Maximum Tolerated Concentration (MTC) of a test article

(1) Concentration group

Experiment 1 group Normal control group

Experiment 2 model control group

Experiment 3 groups example 12 Capsule Contents 125 μ g/mL

Experiment 4 groups example 12 Capsule Contents 250 μ g/mL

Experiment 5 groups example 12 Capsule Contents 500 μ g/mL

Experiment 6 groups example 12 Capsule Contents 1000 μ g/mL

Experiment 7 groups example 12 Capsule Contents 1500 μ g/mL

Experiment 8 groups example 12 Capsule Contents 2000. mu.g/mL

Experiment 9 groups example 11 Capsule Contents 125 μ g/mL

Experiment 10 groups example 11 Capsule Contents 250 μ g/mL

Experiment 11 groups example 11 Capsule Contents 500 μ g/mL

Experiment 12 groups example 11 Capsule Contents 1000. mu.g/mL

Experiment 13 groups example 11 Capsule Contents 1500 μ g/mL

Experiment 14 group of example 11 Capsule Contents 2000. mu.g/mL

(2) Making of models

Treating 2dpf wild type AB strain zebra fish with isoproterenol hydrochloride for 5h, and establishing a zebra fish myocardial injury model.

(3) Experimental methods

Wild type AB strain zebra fish 2 days (2dpf) after fertilization of 780 fish were randomly selected in six-well plates, and 30 zebra fish were treated per well (i.e., per experimental group). The contents 125, 250, 500, 1000, 1500 and 2000. mu.g/mL of the capsules of example 12 and the contents 125, 250, 500, 1000, 1500 and 2000. mu.g/mL of the capsules of example 11 were each administered in water, while the normal control group and the model control group were set to have a volume of 3mL per well. Except for the normal control group, the other experimental groups are all dissolved in water and are given with isoproterenol hydrochloride for induction to establish a zebra fish myocardial injury model. The phenotype and death of zebrafish were observed and recorded after 5h of co-treatment of the contents of the capsules of example 12 and the contents of the capsules of example 11 with isoproterenol hydrochloride, and the MTC of the contents of the capsules of example 12 and the contents of the capsules of example 11 on zebrafish with isoproterenol hydrochloride-induced myocardial injury was determined.

(4) Results of the experiment

See table 1 for details.

The results show that: the zebra fish of the model control group has slight pericardial edema and venous sinus congestion.

Example 12 none of the capsule contents caused death of zebrafish at concentrations of 125 to 2000 μ g/mL. 125. Zebrafish at concentrations of 250, 500 and 1000 μ g/mL all had slight pericardial edema and venous sinus congestion, similar to the model control zebrafish; the zebra fish with the concentration of 1500 mug/mL has venous sinus congestion and the heart rate is slower, and is slightly more serious than the zebra fish of the model control group; the zebra fish with the concentration of 2000 mug/mL has heart rate slowing and body axis bending, and is more serious than the zebra fish of the model control group. Thus, the capsule content of example 12 was determined to have an MTC of 1000 μ g/mL for myocardial-injured zebrafish.

Example 11 the capsule contents did not cause death of zebrafish at concentrations of 125 to 2000 μ g/mL. 125. No obvious abnormality is found in the zebra fish with the concentration of 250 and 500 mu g/mL; the zebra fish with the concentration of 1000 mug/mL has slight pericardial edema and venous sinus congestion, and is similar to the zebra fish of the model control group; the zebra fish with the concentration of 1500 mug/mL has venous sinus congestion which is slightly more serious than the zebra fish of the model control group; zebra fish at a concentration of 2000 μ g/mL had venous sinus congestion and a slower heart rate, which was more severe than the model control zebra fish. Thus, the capsule content of example 11 was determined to have an MTC of 1000 μ g/mL for myocardial injured zebra fish.

TABLE 1 concentration-lethality result of the tested article for treating zebrafish (n as 30)

2.2 experiment two: evaluation of cardiac function-improving Effect of test sample

(1) Concentration group

Experiment 1 group Normal control group

Experiment 2 model control group

Experiment 3 Positive control group

Experiment 4 groups example 12 Capsule Contents 111 μ g/mL

Experiment 5 groups example 12 Capsule Contents 333 μ g/mL

Experiment 6 groups example 12 Capsule Contents 1000 μ g/mL

Experiment 7 groups example 11 Capsule Contents 111 μ g/mL

Experiment 9 groups example 11 Capsule Contents 333 μ g/mL

Experiment 10 groups example 11 Capsule Contents 1000. mu.g/mL

(2) Basis of concentration determination

According to one result of the experiment, the content of the capsule in example 12 has 1000 mug/mL of MTC for the coronary heart disease zebra fish. Therefore, three concentrations of 111. mu.g/mL (1/9MTC), 333. mu.g/mL (1/3MTC) and 1000. mu.g/ML (MTC) were finally selected for evaluation of the cardiac function improvement effect in this experiment.

Example 11 capsule contents MTC of coronary heart disease zebrafish was 1000 μ g/mL. According to the project proposal, three concentrations of 111 mug/mL (1/9MTC), 333 mug/mL (1/3MTC) and 1000 mug/ML (MTC) are finally selected for the cardiac function improvement effect evaluation experiment.

(3) Making of models

Treating 2dpf wild type AB strain zebra fish for 1h by using isoproterenol hydrochloride, and establishing a zebra fish heart failure model.

(4) Experimental methods

225 wild type AB strain zebra fishes 2 days (2dpf) after fertilization were randomly selected in 12-well plates, and 15 zebra fishes were treated per well. The contents 111, 333 and 1000. mu.g/mL of the capsule of example 12, the contents 111, 333 and 1000. mu.g/mL of the capsule of example 11 and the concentration of digoxin, a positive control, were each administered in water, while a normal control and a model control were set to have a volume of 1.5mL per well. Example 12 and example 11 after the capsule content is treated for 4 hours, except for a normal control group, the other experimental groups are dissolved in water and are given with isoproterenol hydrochloride to induce and establish a zebra fish heart failure model. After the contents of the capsules in example 12 and the contents of the capsules in example 11 and isoproterenol hydrochloride are treated for 1 hour, 10 zebra fish in each experimental group are randomly placed under a heartbeat and blood flow analysis system to record zebra fish blood flow videos, the cardiac output (O) and the blood flow velocity (S) of the zebra fish are analyzed and counted, the cardiac output improving effect and the blood flow velocity increasing effect are calculated, and the improving effects of the contents of the capsules in example 12 and the contents of the capsules in example 11 on the heart functions of the heart failure zebra fish are evaluated according to the statistical analysis results. The cardiac output improvement and blood flow velocity increase are calculated as follows:

1) effects on cardiac output;

2) the effect on blood flow velocity;

statistical analysis was performed using analysis of variance and Dunnett's T-test, with p <0.05 indicating significant differences, providing a representative experimental profile.

(5) Results of the experiment

The results show that: the comparison of the cardiac output (0.12nL/s) of the zebra fish in the model control group with the normal control group (0.33nL/s) and the comparison of the blood flow velocity (329 mu m/s) of the zebra fish in the model control group with the normal control group (799 mu m/s) and p <0.001 shows that the model is successfully established. The positive control drug digoxin is 0.8 mug/mL, the cardiac output of the zebra fish is 0.18nL/s, the blood flow rate is 476 mug/s, the increase effects on the cardiac output and the blood flow rate of the zebra fish are respectively 28.6% and 31.3%, and compared with a model control group, the p is less than 0.01, which shows that the digoxin has the improvement effect on the cardiac output and the blood flow rate of the heart failure zebra fish.

Example 12 cardiac output of zebrafish 0.18, 0.21 and 0.20nL/s at concentrations of 111 and 333 and 1000 μ g/mL for capsule contents, respectively, and cardiac output improvement 28.6%, 42.9% and 38.1%, respectively, p <0.05& p <0.001& p <0.01 as compared to model controls; the zebra fish blood flow velocity is 414, 494 and 448 mu m/s respectively, the blood flow velocity increasing effect is 18.1%, 35.1% and 25.3% respectively, and compared with the model control group, p is more than 0.05 and p is less than 0.001 and p is less than 0.05, which shows that under the condition of the experimental concentration, the capsule content in the example 12 has the effect of improving the cardiac output and the blood flow velocity of the heart failure zebra fish.

Example 11 the capsule contents at concentrations of 111 and 333 and 1000 μ g/mL, zebrafish cardiac output was 0.18, 0.15 and 0.16nL/s, respectively, and cardiac output improvement was 27.9%, 14.3% and 19.0%, respectively, p <0.05& p >0.05& p >0.05 as compared to the model control; blood flow velocity of zebra fish was 493, 407 and 349 μm/s, respectively, and blood flow velocity increase was 34.9%, 16.6% and 4.3%, respectively, and p <0.001& p >0.05& p >0.05, compared to the model control group, showed improvement of cardiac output and blood flow velocity of the heart failure zebra fish by the capsule contents of example 11 at a concentration of 111 μ g/mL.

See table 2 for details.

Table 2 zebrafish heart function improvement test results (n ═ 10) for each test group

P <0.05, p <0.01, p <0.001, compared to model controls

2.3 experiment 3: evaluation of myocardial preservation Effect of test article

(1) Concentration group

Experiment 1 group Normal control group

Experiment 2 model control group

Experiment 3 Positive control group

Experiment 4 groups example 12 Capsule Contents 111 μ g/mL

Experiment 5 groups example 12 Capsule Contents 333 μ g/mL

Experiment 6 groups example 12 Capsule Contents 1000 μ g/mL

Experiment 7 groups example 11 Capsule Contents 111 μ g/mL

Experiment 8 groups example 11 Capsule Contents 333 μ g/mL

Experiment 9 groups example 11 Capsule Contents 1000. mu.g/mL

(2) Basis of concentration determination

According to the result of the experiment, the MTC of the content powder of the capsule in the example 12 on the coronary heart disease zebra fish is 1000 mug/mL. Therefore, three concentrations of 111. mu.g/mL (1/9MTC), 333. mu.g/mL (1/3MTC) and 1000. mu.g/ML (MTC) were selected for myocardial preservation evaluation in this experiment.

Example 11 capsule contents MTC of coronary heart disease zebrafish was 1000 μ g/mL. Therefore, three concentrations of 111. mu.g/mL (1/9MTC), 333. mu.g/mL (1/3MTC) and 1000. mu.g/ML (MTC) were selected for myocardial preservation evaluation in this experiment.

(3) Making of models

Treating 2dpf wild type AB strain zebra fish with isoproterenol hydrochloride for 5h, and establishing a zebra fish myocardial injury model.

(4) Experimental methods

Wild type AB strain zebra fish 2 days (2dpf) after fertilization of 450 fish were randomly selected and placed in a six-well plate, and 30 zebra fish were treated in each well. And (3) inducing a zebrafish myocardial injury model by using isoproterenol hydrochloride. The contents 111, 333 and 1000. mu.g/mL of the capsule in example 12, the contents 111, 333 and 1000. mu.g/mL of the capsule in example 11 and the concentration 125. mu.g/mL of the positive control N-acetylcysteine were respectively dissolved in water, and a normal control group and a model control group were set to have a volume of 3mL per well. Example 12 and example 11 the contents of the capsules were treated with isoproterenol hydrochloride for 5 hours, stained with acridine orange (AO for short), photographed after staining and data collection, analyzed and counted for fluorescence intensity (S) of apoptotic cardiac cells of zebra fish and the inhibition rate of myocardial apoptosis was calculated, and the myocardial protection effect of the contents of the capsules of example 12 and example 11 on myocardial-injured zebra fish was evaluated based on the statistical analysis results. The formula for calculating the myocardial protection effect is as follows:

statistical analysis was performed using analysis of variance and Dunnett's T-test, with p <0.05 indicating significant differences, providing a representative experimental profile.

(5) Results of the experiment

The results show that: the comparison of the fluorescence intensity (511206 pixels) of the myocardial apoptotic cells of the zebra fish in the model control group with that of the normal control group (369846 pixels) shows that p is less than 0.001, which indicates that the model is successfully established. The positive control drug N-acetylcysteine zebra fish myocardial apoptosis cell fluorescence intensity is 403264 pixels, the zebra fish myocardial apoptosis inhibition rate is 76%, and p is less than 0.01 compared with a model control group, which shows that N-acetylcysteine has a relatively obvious myocardial protection effect on the zebra fish with myocardial injury.

In example 12, the fluorescence intensities of the apoptotic cell of the myocardium of the zebra fish in the capsule content groups of 111, 333 and 1000 mug/mL are 289677, 341650 and 294153 pixels respectively, the inhibition rates of the apoptotic cell of the zebra fish are 157%, 120% and 154% respectively, and p is less than 0.001 compared with a model control group, which shows that the capsule content in example 12 has obvious myocardial protection effect on the zebra fish with myocardial injury under the condition of the experimental concentration.

Example 11 the fluorescence intensity of the apoptotic cell of the myocardium of zebra fish in the capsule content 111, 333 and 1000 mug/mL concentration groups is 358636, 289761 and 307264 pixels respectively, the inhibition rate of the apoptotic cell of the myocardium of zebra fish is 108%, 157% and 144% respectively, and p of each concentration group is less than 0.001 compared with the model control group, which shows that the capsule content in example 11 has obvious myocardial protection effect on the zebra fish with myocardial injury under the condition of the experimental concentration.

See table 3 for details.

TABLE 3 myocardial preservation of Zebra fish in each experimental group

P <0.05, p <0.01, p <0.001, compared to model controls

The above experimental results also prove that the two traditional Chinese medicine compositions provided by the invention play a key and irreplaceable role in protecting cardiac muscle and improving cardiac function.

In a word, the invention provides a traditional Chinese medicine composition for treating ischemic cardiomyopathy.

The composition comprises folium Pyrrosiae, folium Ginkgo, caulis Perillae, herba Equiseti hiemalis, rhizoma Polygoni Cuspidati, herba abri, and optionally radix Puerariae. The composition of the invention has the effects of expelling pathogenic factors and relieving muscles, promoting qi and activating blood circulation by combining various medicines, cutting the pathogenesis of ischemic cardiomyopathy and treating both principal and secondary aspects of disease. Pharmacological experiments prove that the traditional Chinese medicine composition has extremely obvious protective effect on cardiac muscle of zebra fish with myocardial injury and has the effect of improving the cardiac function of the zebra fish with heart failure. The invention provides a new clinical treatment option for patients with ischemic cardiomyopathy and doctors.