Traditional Chinese medicine composition for treating myocardial infarction, medicinal preparation and preparation method thereof

Abstract

The invention provides a traditional Chinese medicine composition for treating myocardial infarction, a medicinal preparation and a preparation method thereof, and belongs to the technical field of medicines. The traditional Chinese medicine composition comprises the following components: astragalus membranaceus, codonopsis pilosula, ligusticum wallichii, salvia miltiorrhiza, lily, lindera aggregata, radix rehmanniae, pseudo-ginseng and rhizoma cyperi. The traditional Chinese medicine composition and the medicinal preparation provided by the invention have the effects of tonifying qi and activating blood, nourishing yin and dredging collaterals, and promoting qi circulation and relieving pain, and can be used for treating qi-yin deficiency and blood stasis syndromes such as myocardial infarction accompanied by shortness of breath and weakness, chest pain such as stabbing, dysphoria and insomnia, dry mouth and constipation.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a traditional Chinese medicine composition for treating myocardial infarction, a pharmaceutical preparation, and a method for preparing the same. Background Technology

[0002] Myocardial infarction, also known as a heart attack, is a life-threatening disease caused by acute blockage of the coronary arteries, leading to insufficient blood supply to the corresponding area of ​​the myocardium and resulting in myocardial necrosis. This disease is mainly caused by multiple factors, including insufficient blood supply, myocardial hypoxia, and excessive myocardial oxygen consumption, resulting from various causes such as coronary atherosclerosis, arrhythmia, and respiratory failure. Globally, acute myocardial infarction has a high incidence and mortality rate, and is more common in middle-aged and elderly people, as well as high-risk groups such as those with heart disease, smokers, diabetics, and those with high cholesterol.

[0003] The primary goal of treating acute myocardial infarction is to immediately open the coronary arteries and slow the progression of myocardial ischemia and infarction. Common treatment methods include drug therapy, such as the use of antiplatelet drugs, anticoagulants, and lipid-lowering drugs, as well as interventional and surgical treatments, such as percutaneous coronary intervention and emergency coronary artery bypass grafting. The main ways to prevent myocardial infarction are to control risk factors for heart disease such as hypertension, diabetes, and hyperlipidemia, and to maintain a healthy lifestyle, such as quitting smoking, engaging in moderate exercise, and maintaining a balanced diet.

[0004] Traditional Chinese medicine (TCM) treatment for myocardial infarction varies depending on the underlying cause and symptoms. For example, Yixin Granules contain ginseng, Ophiopogon japonicus, Schisandra chinensis, Anemarrhena asphodeloides, Angelica sinensis, and Acorus tatarinowii; its effects focus on tonifying Qi and nourishing Yin, supplemented by promoting blood circulation and unblocking the meridians; it is suitable for stable angina pectoris, with symptoms such as chest tightness, palpitations, shortness of breath, insomnia, and excessive sweating; this medicine is warming and tonifying, with a simple formula, but its Yin-nourishing effect is limited. Tongqiao Yixin Pills contain artificial musk, bezoar, toad venom, borneol, Panax notoginseng, ginseng, and buffalo horn; its effects focus on promoting blood circulation, removing blood stasis, opening the orifices, relieving pain, and improving myocardial blood supply; it is suitable for acute chest pain attacks (such as angina pectoris), palpitations, and shortness of breath, especially suitable for coronary heart disease with significant blood stasis; this medicine contains potent orifice-opening ingredients such as toad venom and borneol, and its strong medicinal properties make it suitable for acute symptoms. Compound Danshen Dripping Pills contain Danshen, Panax notoginseng, and borneol. Its effects focus on promoting blood circulation, removing blood stasis, regulating qi, relieving pain, and improving myocardial ischemia. It is suitable for patients with stable angina pectoris, acute chest pain, and obvious blood stasis syndrome in coronary heart disease. This medicine is specifically designed to promote blood circulation, remove blood stasis, regulate qi, and relieve pain. It is mainly for acute symptoms and has the characteristics of rapid and high efficacy. Sublingual administration can quickly relieve symptoms.

[0005] Since the symptoms of myocardial infarction vary depending on the cause, and different traditional Chinese medicines have different focuses in their efficacy, more traditional Chinese medicines for treating myocardial infarction still need to be developed in order to better address different types of diseases. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the present invention aims to provide a traditional Chinese medicine composition, pharmaceutical preparation, and preparation method for treating myocardial infarction. This traditional Chinese medicine composition and pharmaceutical preparation possess the effects of invigorating qi and promoting blood circulation, nourishing yin and unblocking collaterals, and relieving pain. It can be used to treat myocardial infarction accompanied by shortness of breath, fatigue, stabbing chest pain, irritability, insomnia, dry mouth, and constipation, which are symptoms of qi and yin deficiency with blood stasis.

[0007] To achieve this objective, the present invention adopts the following technical solution:

[0008] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating myocardial infarction, the traditional Chinese medicine composition comprising the following components:

[0009] Astragalus, Codonopsis, Ligusticum chuanxiong, Salvia miltiorrhiza, Lilium brownii, Lindera strychnifolia, Rehmannia glutinosa, Panax notoginseng, and Cyperus rotundus.

[0010] In some preferred embodiments of the present invention, the traditional Chinese medicine composition comprises the following components in parts by weight:

[0011] Astragalus membranaceus 22-26 parts, Codonopsis pilosula 18-23 parts, Ligusticum chuanxiong 7-11 parts, Salvia miltiorrhiza 13-17 parts, Lilium brownii 13-18 parts, Lindera strychnifolia 8-12 parts, Rehmannia glutinosa 18-23 parts, Panax notoginseng 10-14 parts, and Cyperus rotundus 7-12 parts.

[0012] The weight of Astragalus membranaceus can be any value within the range of 22-26 parts, such as 22 parts, 22.2 parts, 22.5 parts, 22.8 parts, 23 parts, 23.2 parts, 23.5 parts, 23.8 parts, 24 parts, 24.2 parts, 24.5 parts, 24.8 parts, 25 parts, 25.2 parts, 25.5 parts, 25.8 parts, or 26 parts, etc.

[0013] The weight of Codonopsis pilosula can be any value within the range of 18-23 parts, such as 18 parts, 18.2 parts, 18.5 parts, 18.8 parts, 19 parts, 19.2 parts, 19.5 parts, 19.8 parts, 20 parts, 20.2 parts, 20.5 parts, 20.8 parts, 21 parts, 21.2 parts, 21.5 parts, 21.8 parts, 22 parts, 22.2 parts, 22.5 parts, 22.8 parts, or 23 parts, etc.

[0014] The weight of Chuanxiong can be any value within the range of 7-11 parts, such as 7 parts, 7.2 parts, 7.5 parts, 7.8 parts, 8 parts, 8.2 parts, 8.5 parts, 8.8 parts, 9 parts, 9.2 parts, 9.5 parts, 9.8 parts, 10 parts, 10.2 parts, 10.5 parts, 10.8 parts, or 11 parts, etc.

[0015] The weight of Danshen can be any value within the range of 13-17 parts, such as 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts, or 17 parts, etc.

[0016] The weight of lilies can be any value within the range of 13-18 parts, such as 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, 14 parts, 14.2 parts, 14.5 parts, 14.8 parts, 15 parts, 15.2 parts, 15.5 parts, 15.8 parts, 16 parts, 16.2 parts, 16.5 parts, 16.8 parts, 17 parts, 17.2 parts, 17.5 parts, 17.8 parts, or 18 parts, etc.

[0017] The weight of Lindera root can be any value within the range of 8-12 parts, such as 8 parts, 8.2 parts, 8.5 parts, 8.8 parts, 9 parts, 9.2 parts, 9.5 parts, 9.8 parts, 10 parts, 10.2 parts, 10.5 parts, 10.8 parts, 11 parts, 11.2 parts, 11.5 parts, 11.8 parts, or 12 parts, etc.

[0018] The weight percentage of raw rehmannia root can be any value within the range of 18-23 parts, such as 18 parts, 18.2 parts, 18.5 parts, 18.8 parts, 19 parts, 19.2 parts, 19.5 parts, 19.8 parts, 20 parts, 20.2 parts, 20.5 parts, 20.8 parts, 21 parts, 21.2 parts, 21.5 parts, 21.8 parts, 22 parts, 22.2 parts, 22.5 parts, 22.8 parts, or 23 parts, etc.

[0019] The weight of Panax notoginseng can be any value within the range of 10-14 parts, such as 10 parts, 10.2 parts, 10.5 parts, 10.8 parts, 11 parts, 11.2 parts, 11.5 parts, 11.8 parts, 12 parts, 12.2 parts, 12.5 parts, 12.8 parts, 13 parts, 13.2 parts, 13.5 parts, 13.8 parts, or 14 parts, etc.

[0020] The weight of Cyperus rotundus can be any value within the range of 7-12 parts, such as 7 parts, 7.2 parts, 7.5 parts, 7.8 parts, 8 parts, 8.2 parts, 8.5 parts, 8.8 parts, 9 parts, 9.2 parts, 9.5 parts, 9.8 parts, 10 parts, 10.2 parts, 10.5 parts, 10.8 parts, 11 parts, 11.2 parts, 11.5 parts, 11.8 parts, or 12 parts, etc.

[0021] In some preferred embodiments of the present invention, the traditional Chinese medicine composition comprises the following components in parts by weight:

[0022] Astragalus membranaceus 24 parts, Codonopsis pilosula 20 parts, Ligusticum chuanxiong 9 parts, Salvia miltiorrhiza 15 parts, Lilium brownii 15 parts, Lindera strychnifolia 10 parts, Rehmannia glutinosa 20 parts, Panax notoginseng 12 parts, and Cyperus rotundus 9 parts.

[0023] The efficacy of each component in the traditional Chinese medicine composition provided by this invention is as follows:

[0024] Astragalus: Tonifies Qi and raises Yang, consolidates the exterior and stops sweating;

[0025] Codonopsis pilosula: replenishes qi and strengthens the spleen and lungs;

[0026] Chuanxiong: invigorates blood circulation, promotes qi circulation, dispels wind, and relieves pain;

[0027] Danshen: Promotes blood circulation, removes blood stasis, regulates menstruation, and relieves pain;

[0028] Lily: Nourishes Yin and moistens the lungs, calms the mind and soothes the nerves;

[0029] Lindera root: promotes qi circulation and relieves pain, warms the kidneys and dispels cold;

[0030] Rehmannia glutinosa: Clears heat and cools the blood, nourishes yin and promotes the production of body fluids;

[0031] Panax notoginseng: disperses blood stasis, stops bleeding, reduces swelling, and relieves pain;

[0032] Cyperus rotundus: Soothes the liver and regulates qi, regulates menstruation and relieves pain.

[0033] The compatibility mechanism of each component is as follows:

[0034] The principal herbs are Astragalus membranaceus and Codonopsis pilosula. Astragalus membranaceus tonifies Qi and raises Yang, while Codonopsis pilosula strengthens the spleen and benefits the lungs. Together, they greatly replenish vital energy and are essential herbs for treating Qi deficiency, specifically addressing the underlying pathogenesis of myocardial infarction.

[0035] Assistant herbs: Chuanxiong (Ligusticum striatum), Danshen (Salvia miltiorrhiza), and Sanqi (Panax notoginseng). Chuanxiong invigorates blood and promotes qi circulation, Danshen invigorates blood and removes blood stasis, and Sanqi disperses blood stasis and stops bleeding. When used together, these three herbs invigorate blood and remove blood stasis, improve myocardial blood supply, and assist the principal herbs in treating the symptoms of qi stagnation and blood stasis.

[0036] Adjuvant herbs: lily bulb, rehmannia root, lindera root, and cyperus rhizome. Lily bulb moistens the lungs and clears the heart, while rehmannia root clears heat and cools the blood. Both nourish yin and moisten dryness, preventing the blood-activating herbs from damaging yin. Lindera root promotes qi circulation and relieves pain, while cyperus rhizome soothes the liver and regulates qi, enhancing the efficacy of the blood-activating herbs.

[0037] Guiding herb: Cyperus rotundus serves as a guiding herb, directing the medicine into the liver meridian and regulating the flow of qi and blood.

[0038] The traditional Chinese medicine composition provided by this invention focuses on tonifying qi and promoting blood circulation. It integrates the qi-tonifying and meridian-clearing principles of Buyang Huanwu Decoction and the blood-activating and stasis-removing principles of Xuefu Zhuyu Decoction, and adds yin-nourishing and qi-regulating herbs to harmonize its warming and drying properties. Its characteristics are "tonifying without stagnation and activating without harming", making it suitable for chronic diseases with mixed deficiency and excess patterns, especially for common modern metabolic and cardiovascular diseases.

[0039] The traditional Chinese medicine composition provided by this invention has the effects of invigorating qi and blood, nourishing yin and unblocking collaterals, and promoting qi circulation and relieving pain. It can be used to treat myocardial infarction accompanied by shortness of breath, fatigue, sharp chest pain, irritability, insomnia, dry mouth and constipation, which are symptoms of qi and yin deficiency with blood stasis. For example, it can be used to treat acute ST-segment elevation myocardial infarction (qi deficiency and blood stasis syndrome) after PCI (percutaneous coronary intervention) or bypass surgery.

[0040] In a second aspect, the present invention provides a pharmaceutical preparation for treating myocardial infarction, the pharmaceutical preparation comprising the effective components of the traditional Chinese medicine composition as described in the first aspect.

[0041] In some embodiments of the present invention, the pharmaceutical preparation further includes excipients.

[0042] In some embodiments of the present invention, the dosage form of the pharmaceutical preparation is granules, tablets, capsules, or decoction.

[0043] Thirdly, the present invention provides a method for preparing a pharmaceutical preparation as described in the second aspect, the method comprising the following steps:

[0044] (1) The traditional Chinese medicine composition described in the first aspect is subjected to water extraction to obtain an aqueous extract;

[0045] (2) The aqueous extract is prepared into the dosage form of the drug preparation.

[0046] In some embodiments of the present invention, the water extraction step includes:

[0047] The herbal composition is decocted with water at least twice (e.g., twice, three times, four times, or five times), each time for 1-4 hours (e.g., 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, or 4 hours). After each decoction, the mixture is filtered, and the filtrates are combined to obtain the aqueous extract.

[0048] In some embodiments of the present invention, in the water extraction step, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition; for example, it can be 18 times, 19 times, 20 times, 22 times, 23 times, 25 times, 26 times, 28 times, 30 times, 32 times, 33 times or 35 times, etc.

[0049] In some embodiments of the present invention, the water extraction step involves two decoctions. For the first decoction, the mass of water is 10-20 times the mass of the herbal composition (e.g., 10, 12, 13, 15, 16, 18, or 20 times, etc.), and the decoction time is 2-3 hours (e.g., 2 hours, 2.2 hours, 2.3 hours, 2.5 hours, 2.6 hours, 2.8 hours, or 3 hours, etc.). For the second decoction, the mass of water is 8-15 times the mass of the herbal composition (e.g., 8, 9, 10, 11, 12, 13, 14, or 15 times, etc.), and the decoction time is 1-2 hours (e.g., 1 hour, 1.2 hours, 1.3 hours, 1.5 hours, 1.6 hours, 1.8 hours, or 2 hours, etc.).

[0050] Compared with the prior art, the present invention has the following beneficial effects:

[0051] This invention utilizes a scientific combination of Astragalus membranaceus, Codonopsis pilosula, Ligusticum chuanxiong, Salvia miltiorrhiza, Lilium brownii, Lindera strychnifolia, Rehmannia glutinosa, Panax notoginseng, and Cyperus rotundus to obtain a traditional Chinese medicine composition and pharmaceutical preparation that has the effects of invigorating qi and blood, nourishing yin and unblocking collaterals, and promoting qi circulation and relieving pain. It can be used to treat myocardial infarction accompanied by shortness of breath and fatigue, stabbing chest pain, irritability and insomnia, dry mouth and constipation, etc., which are symptoms of qi and yin deficiency with blood stasis. Attached Figure Description

[0052] Figure 1 Diagrams showing the sham surgery and modeling process in rats;

[0053] Figure 2A Representative electrocardiograms of rats before and after modeling;

[0054] Figure 2B This is a comparison of ST segment values ​​in rats before and after modeling.

[0055] Figure 3A A bar chart showing the CO levels of rats in each group;

[0056] Figure 3B EF bar chart for rats in each group;

[0057] Figure 3C FS bar chart for rats in each group;

[0058] Figure 3D Bar chart showing LVIDs of rats in each group;

[0059] Figure 3E The bar chart shows the LVIDd values ​​of rats in each group;

[0060] Figure 4A Bar chart showing the cardiac weight index of rats in each group;

[0061] Figure 4B Bar chart showing the cardiotibial ratio of rats in each group;

[0062] Figure 5A A bar chart showing the serum CK levels in each group of rats;

[0063] Figure 5B Bar chart showing serum CK-MB levels in rats of each group;

[0064] Figure 6A Bar chart showing serum cTnT levels in rats of each group;

[0065] Figure 6B Bar chart showing serum MMP-9 levels in rats of each group;

[0066] Figure 7 HE staining images of heart tissue from each group of rats. Detailed Implementation

[0067] The technical solution of the present invention will be further described below with reference to the accompanying drawings and specific embodiments. Those skilled in the art should understand that the specific embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention.

[0068] Example 1

[0069] This embodiment provides a traditional Chinese medicine composition for treating myocardial infarction, comprising the following components in parts by weight:

[0070] Astragalus membranaceus 22 parts, Codonopsis pilosula 23 parts, Ligusticum chuanxiong 7 parts, Salvia miltiorrhiza 17 parts, Lilium brownii 13 parts, Lindera strychnifolia 12 parts, Rehmannia glutinosa 18 parts, Panax notoginseng 14 parts, and Cyperus rotundus 7 parts.

[0071] This embodiment also provides a decoction for treating myocardial infarction, the preparation method of which is as follows:

[0072] Weigh the medicinal materials according to the proportions of the Chinese herbal composition, add water and decoct twice. For the first decoction, add 20 times the amount of water as the medicinal materials and decoct for 3 hours. For the second decoction, add 10 times the amount of water as the medicinal materials and decoct for 2 hours. After each decoction, filter, combine the filtrates, and concentrate under reduced pressure to obtain a decoction for treating myocardial infarction.

[0073] Example 2

[0074] This embodiment provides a traditional Chinese medicine composition for treating myocardial infarction, comprising the following components in parts by weight:

[0075] Astragalus membranaceus 26 parts, Codonopsis pilosula 18 parts, Ligusticum chuanxiong 11 parts, Salvia miltiorrhiza 13 parts, Lilium brownii 18 parts, Lindera strychnifolia 8 parts, Rehmannia glutinosa 23 parts, Panax notoginseng 10 parts, and Cyperus rotundus 12 parts.

[0076] This embodiment also provides a decoction for treating myocardial infarction, the preparation method of which is as follows:

[0077] Weigh the medicinal materials according to the ratio of the traditional Chinese medicine composition, decoct them with water twice. For the first decoction, add 10 times the mass of the medicinal materials in water and decoct for 2 hours; for the second decoction, add 8 times the mass of the medicinal materials in water and decoct for 1 hour. Filter after each decoction, combine the filtrates, and obtain the decoction for treating myocardial infarction after concentration under reduced pressure.

[0078] Example 3

[0079] This example provides a traditional Chinese medicine composition for treating myocardial infarction, including the following components in parts by weight:

[0080] 24 parts of Astragalus membranaceus, 20 parts of Codonopsis pilosula, 9 parts of Ligusticum chuanxiong, 15 parts of Salvia miltiorrhiza, 15 parts of Lilium brownii, 10 parts of Lindera aggregata, 20 parts of Rehmannia glutinosa, 12 parts of Panax notoginseng, and 9 parts of Cyperus rotundus.

[0081] This example also provides a decoction for treating myocardial infarction, and its preparation method is as follows:

[0082] Weigh the medicinal materials according to the ratio of the traditional Chinese medicine composition, decoct them with water twice. For the first decoction, add 15 times the mass of the medicinal materials in water and decoct for 2.5 hours; for the second decoction, add 15 times the mass of the medicinal materials in water and decoct for 1.5 hours. Filter after each decoction, combine the filtrates, and concentrate under reduced pressure to 3.49 g of traditional Chinese medicine composition / mL to obtain the decoction for treating myocardial infarction, and store it at 4°C for later use.

[0083] Therapeutic efficacy test

[0084] Use the decoction for treating myocardial infarction prepared in Example 3 for animal experiments to confirm its therapeutic effect on the rat myocardial infarction model.

[0085] 1. Test materials

[0086] 1.1. Test animals

[0087] SPF-grade SD rats, male, weighing 250 - 260 g, are used in the test. They are purchased from Spf (Suzhou) Biotechnology Co., Ltd., with the production license number SCXK (Su) 2022 - 0006 and the certificate number A202412100045. The feeding environmental conditions: The feeding environmental conditions are standard, referring to the national standard GB14925 - 2010 of the People's Republic of China; the temperature is 20 - 26°C (daily temperature difference ≤ 4°C); the relative humidity is 40 - 70%; the pressure in the feeding room ≥ 10 Pa; 12 / 12 hour day-night light-dark alternation.

[0088] 1.2. Test reagents

[0089] The decoction for treating myocardial infarction prepared in Example 3 is diluted with sterile injection water to 6.98 g / 8 mL, 13.96 g / 8 mL, and 27.92 g / 8 mL respectively. This concentration refers to the mass of the traditional Chinese medicine composition corresponding to the decoction contained in each 8 mL of the solution.

[0090] Tongxinluo Capsules, specification 0.26g / capsule, batch number A2408009, sealed, store in a cool place, manufacturer: Shijiazhuang Yiling Pharmaceutical Co., Ltd.; product approval number: National Drug Approval Number Z19980015. Preparation method: Dilute the contents of Tongxinluo Capsules with sterile water for injection to 0.33g / 8mL.

[0091] Sterile water for injection, 500 mL / bottle, batch number 240621, to be stored in a sealed container, manufactured by Shaanxi Sheng'ao Animal Pharmaceutical Co., Ltd.; product approval number is Veterinary Drug No. 270071791.

[0092] This is a general-purpose tissue fixative, available in 500mL vials, batch number GP24103081634. Store at room temperature. The manufacturer is Wuhan Saiweier Biotechnology Co., Ltd.

[0093] 1.3. Instruments

[0094] The pipette is manufactured by Eppendorf and has a model number of 200 μl.

[0095] The electronic balance is manufactured by Shanghai Yaoxin Electronic Technology Co., Ltd., and its model number is LQ-C12001.

[0096] The benchtop high-speed refrigerated microcentrifuge is manufactured by Thermo and its model number is Sorvall Legand Micro 17R.

[0097] Mini mixing centrifuge, manufacturer: LABGIC, model: L-CM-MINI;

[0098] The microplate reader is manufactured by Thermo and is model VARIOSKAN LUX.

[0099] The dehydrator is manufactured by DIAPATH and its model is Donatello.

[0100] The embedding machine is manufactured by Wuhan Junjie Electronics Co., Ltd., and its model number is JB-P5.

[0101] The freezing station is manufactured by Wuhan Junjie Electronics Co., Ltd., and its model number is JB-L5.

[0102] The pathology slider is manufactured by Leica Instruments Shanghai Co., Ltd., and its model number is RM2016.

[0103] The sheet spreader is manufactured by Zhejiang Jinhua Kedi Instrument Equipment Co., Ltd., and its model is KD-P.

[0104] The oven is manufactured by Tianjin Laiborui Instrument Equipment Co., Ltd., and its model is GFL-230.

[0105] An upright optical microscope, manufactured by Nikon Japan, model Nikon Eclipse E100;

[0106] The imaging system is manufactured by Nikon of Japan, and its model is NIKON DS-U3.

[0107] Upright white light photographic microscope, manufactured by Nikon Japan, model Eclipse Ci-L;

[0108] The electrocardiograph (ECG) machine is manufactured by Nanjing Calvin Biotechnology Co., Ltd., model Medlab, and device number KW2024092704.

[0109] The dual-channel small animal ventilator is manufactured by Shanghai Aulcot Biotechnology Co., Ltd., and its model number is ALC-V8-SLA.

[0110] 2. Test Methods

[0111] 2.1. Modeling, grouping, and drug administration

[0112] The day before modeling, rats were fasted but allowed free access to water, fasting for 12 hours. On the second day, the rat myocardial infarction model was established: rats were anesthetized, their skin was prepared using a small pet shaver, and they were fixed on an animal operating table. After disinfection with iodine, a sterile drape was applied. The location of the rat's heart (3rd-4th intercostal space) was determined, and the skin was cut open. The pectoral muscle layers were bluntly dissected layer by layer to expose the heart. A sterile suture needle was used to ligate the artery and vein at the left anterior descending coronary artery. After stabilization, an electrocardiograph was used to record electrocardiograms in the limb leads. The presence of J-point deviation or elevation, and tall T waves indicated myocardial ischemia and myocardial infarction. Visual observation showed the ischemic myocardium to be dark red, indicating successful ligation. The intercostal muscles and skin were sutured layer by layer. Postoperatively, 200,000 U / day of penicillin sodium was injected intramuscularly for 3 consecutive days to prevent infection. A portion of the rats underwent sham surgery: only sutures were threaded through the blood vessels in the myocardial tissue without ligation. The rat sham surgery and modeling process are illustrated below. Figure 1 As shown, A represents sham surgery, and B represents the model. Representative electrocardiograms of rats before and after modeling are shown below. Figure 2A As shown in the figure. The comparison of ST segment values ​​in rats before and after modeling is shown in the figure. Figure 2B As shown; among them, compared with the preoperative period, *** p < 0.001. During modeling, gross observation showed that the myocardial tissue at the ligation site of the left coronary artery gradually turned white, the ST segment of lead II of the electrocardiogram showed obvious upward convexity, and at the same time, the local color of the left ventricle darkened and the vascular pulsation weakened.

[0113] Rats surviving 24 hours after modeling were divided into five groups: model group, low-dose prescription group, medium-dose prescription group, high-dose prescription group, and Tongxinluo capsule group, with eight rats in each group. A sham-operated group consisted of ten rats. Each group of rats received the medication orally twice daily for 39 consecutive days. The types and dosages of medication administered to each group are as follows:

[0114] Sham surgery group and model group: normal saline, administration volume 800μL / 100g;

[0115] Low-dose prescription group: a diluted solution of the decoction prepared in Example 3 with a concentration of 6.98 g / 8 mL, administered at a dose of 6.98 g / kg and a volume of 800 μL / 100 g;

[0116] The dosage group in the prescription was a diluted solution of the decoction prepared in Example 3 with a concentration of 13.96 g / 8 mL, with a dosage of 13.96 g / kg and a dosage volume of 800 μL / 100 g.

[0117] High-dose prescription group: a diluted solution of the decoction prepared in Example 3 with a concentration of 27.92 g / 8 mL, administered at a dose of 27.92 g / kg, with an administration volume of 800 μL / 100 g;

[0118] Tongxinluo capsule group: a diluted solution of Tongxinluo capsules with a concentration of 0.33g / 8mL, a dosage of 0.33g / kg, and a dosage volume of 800μL / 100g.

[0119] 2.2. Detection Indicators and Methods

[0120] 2.2.1. Electrocardiogram

[0121] The electrocardiogram (ECG) was used for testing.

[0122] 2.2.2. Echocardiography

[0123] Echocardiography was performed on rats in each group before sampling. Rats were anesthetized with gas, fixed in a dorsal position, and extensively prepared on the left anterior chest to induce a sustained and stable heart rate. High-resolution small animal ultrasound was used for echocardiography, with the ultrasound personnel blinded. The rats were tilted 30° to the left and ultrasound coupling gel was applied. The ultrasound probe was placed on the left side of the sternum at a 10°-30° angle to the midline of the sternum to display the long-axis view of the left ventricle. M-mode ultrasound was applied to the long-axis view of the left ventricle, allowing the sampling line to pass through the left anterior and posterior walls at the level of the mitral valve chordae tendineae. The probe was then rotated 90° clockwise and adjusted appropriately to display the short-axis view of the left ventricle. Anatomical M-mode ultrasound was used on the short-axis view of the left ventricular papillary muscles, allowing the sampling line to pass through the anterior septum and posterior wall, guided by a two-dimensional image. The M-mode curve was recorded and measured to capture the left ventricular motion curve. Cardiac output (CO), left ventricular end-systolic diameter (LVIDS), left ventricular end-diastolic diameter (LVIDd), left ventricular ejection fraction (LVEF), and left ventricular fractional shortening (LVFS) were measured, with at least 3 heartbeat cycles measured for each rat.

[0124] 2.2.3. HE staining of cardiac tissue

[0125] (1) Paraffin slice preparation

[0126] 1) Sample collection: After the experiment, the rats were euthanized, and fresh heart tissue was collected and fixed with fixative for more than 24 hours. The tissue was removed from the fixative and trimmed in a fume hood using a scalpel. The trimmed tissue and corresponding labels were placed in a dehydration box.

[0127] 2) Dehydration and wax impregnation: The dehydration box is placed in a dehydrator for gradient alcohol dehydration. The steps are as follows: 75% alcohol for 4 hours, 85% alcohol for 2 hours, 90% alcohol for 2 hours, 95% alcohol for 1 hour, anhydrous ethanol I for 30 minutes, anhydrous ethanol II for 30 minutes, benzyl alcohol for 10 minutes, xylene I for 10 minutes, xylene II for 10 minutes, 65℃ melted paraffin I for 1 hour, 65℃ melted paraffin II for 1 hour, and 65℃ melted paraffin III for 1 hour.

[0128] 3) Embedding: The paraffin-impregnated tissue is embedded in an embedding machine. First, the molten paraffin is placed into the embedding frame. Before the paraffin solidifies, the tissue is removed from the dehydration box, placed into the embedding frame according to the embedding surface requirements, and labeled accordingly. The tissue is then cooled on a -20°C freezing stage. After the paraffin solidifies, the paraffin block is removed from the embedding frame and trimmed.

[0129] 4) Sectioning: Place the trimmed wax block on a -20℃ freezing stage to cool, then place the cooled wax block on a paraffin microtome to section to a thickness of 4μm. Float the sections on 40℃ warm water in a slide to flatten the tissue, then lift the tissue onto a glass slide and bake in a 60℃ oven. After the wax has melted in the water, remove the slide and store it at room temperature for later use.

[0130] (2) Staining

[0131] 1) Dewaxing paraffin sections to water: Place the sections in xylene I for 20 min, xylene II for 20 min, anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, 75% ethanol for 5 min, and then wash with tap water.

[0132] 2) Hematoxylin staining: After dewaxing, the sections were stained with hematoxylin solution for 5 minutes, washed with tap water, differentiated with differentiation solution, washed with tap water, blued with blue solution, and rinsed with running water.

[0133] 3) Eosin staining: After hematoxylin staining, the sections were dehydrated in 85% and 95% alcohol for 5 minutes each, and then stained in eosin staining solution for 5 minutes.

[0134] 4) Dehydration and mounting: After eosin staining, the sections were sequentially immersed in anhydrous ethanol I for 5 min, anhydrous ethanol II for 5 min, anhydrous ethanol III for 5 min, xylene I for 5 min, xylene II for 5 min, and then mounted with clear neutral resin.

[0135] 5) Microscopic examination, image acquisition and analysis.

[0136] 2.2.4. Heart weight index, tibia ratio

[0137] After the experiment, the rats were euthanized, and their hearts and tibias were removed. The hearts were weighed, and the tibia length was measured. The heart-to-tibia ratio was calculated using the following formulas:

[0138] Heart weight index = heart weight / rat body weight × 100%;

[0139] Heart-to-tibia ratio = heart weight / rat tibia length.

[0140] 2.2.5. Serum CK and CK-MB levels

[0141] After anesthetizing rats, blood was collected from the abdominal aorta. After standing at room temperature for 2 hours, the blood was centrifuged at 3000 rpm for 15 minutes at 4℃. The supernatant was collected and tested on an automated biochemical analyzer after setting the appropriate parameters.

[0142] 2.2.6. Serum cTnT and MMP-9 levels

[0143] The detection was performed using an ELISA kit, and the steps are as follows:

[0144] (1) Sample collection: Use test tubes free of pyrogens and endotoxins, and avoid any cell stimulation during the operation. After collecting blood, centrifuge at 3000 rpm for 10 minutes to quickly and carefully separate serum and red blood cells.

[0145] (2) Remove the required strips from the aluminum foil bag after equilibration at room temperature for 20 minutes, and seal the remaining strips in a self-sealing bag and return them to 4℃.

[0146] (3) Set up standard wells and sample wells, and add 50 μL of standard of different concentrations to each standard well.

[0147] (4) Add 10 μL of the sample to be tested to the sample well first, and then add 40 μL of sample diluent; do not add to the blank well.

[0148] (5) Except for the blank wells, add 100 μL of horseradish peroxidase (HRP) labeled detection antibody to each of the standard wells and sample wells, seal the reaction wells with sealing film, and incubate at 37°C in a water bath or incubator for 60 min.

[0149] (6) Discard the liquid, pat dry on absorbent paper, fill each well with washing liquid, let stand for 1 minute, shake off the washing liquid, pat dry on absorbent paper, and repeat the washing process 5 times (or use a plate washer).

[0150] (7) Add 50 μL of substrate A and B to each well and incubate at 37°C in the dark for 15 min.

[0151] (8) Add 50 μL of stop solution to each well and measure the OD value of each well at a wavelength of 450 nm within 15 min.

[0152] (9) Plot the standard curve: plot the linear regression curve of the standard with the concentration of the standard on the x-axis and the corresponding OD value on the y-axis, and calculate the concentration value of each sample according to the curve equation.

[0153] 2.3. Data Analysis and Processing

[0154] GraphPad 9.5.0 and SPSS 26 software were used for plotting and statistical analysis. Data are expressed as mean ± SEM. One-way ANOVA was used for statistical analysis to test for significant differences between the two groups. P < 0.05 was considered statistically significant.

[0155] 3. Test Results

[0156] 3.1. Effects of the formulation of this invention on echocardiographic parameters in rats with myocardial infarction

[0157] Figure 3A The bar chart shows the cardiac output (CO) of rats in each group. Figure 3BThe bar chart shows the EF (left ventricular ejection fraction) of rats in each group. Figure 3C The bar chart shows the FS (left ventricular fractional shortening) of rats in each group. Figure 3D The bar chart shows the LVIDs (left ventricular end-systolic diameter) of rats in each group. Figure 3E The bar chart shows the LVIDd (left ventricular end-diastolic diameter) of rats in each group. Compared with the sham-operated group, ** p < 0.01, *** p < 0.001; compared with the model group, # p < 0.05 ## p < 0.01, ### p < 0.001; compared with the Tongxinluo capsule group, @ p < 0.05 @@@ p < 0.001.

[0158] Figures 3A-3E The results showed that, compared with the sham-operated group, the model group rats had significantly increased left ventricular end-systolic diameter (LVIDs) and left ventricular end-diastolic diameter (LVIDd) (P<0.01, P<0.001), significantly decreased left ventricular ejection fraction (EF) and left ventricular fractional shortening (FS) (P<0.001, P<0.001), and significantly decreased cardiac output (CO) (P<0.001). After administration, compared with the model group rats, the rats in each dose group of the traditional Chinese medicine prescription of this invention had decreased LVIDs and LVIDd (P<0.001), and increased EF, FS, and CO (P<0.05, P<0.01, P<0.001). Compared with the model group rats, the rats in the Tongxinluo capsule group had increased EF, FS, and CO (P<0.05, P<0.01, P<0.001), but LVIDs and LVIDd did not change significantly (P>0.05). Compared with the Tongxinluo capsule group, the rats in each dosage group of the traditional Chinese medicine prescription of this invention showed a more significant decrease in LVIDs (P<0.01), but the increase in CO was not as significant as that in the Tongxinluo capsule group (P<0.05).

[0159] 3.2. Effects of the formulation of this invention on cardiac weight index and heart-to-tibia ratio in rats with myocardial infarction

[0160] Figure 4A The bar chart shows the cardiac weight index of rats in each group. Figure 4B The bar chart shows the cardiotibial ratio of rats in each group. Compared to the sham-operated group, ** p < 0.01, *** p < 0.001; compared with the model group, # p < 0.05 ## p < 0.01, ### p < 0.001.

[0161] from Figure 4Aand Figure 4B It can be seen that, compared with the sham-operated group, the cardiac weight index and cardiac-to-tibia ratio of the model group rats were significantly increased (P<0.01, P<0.001). After administration, compared with the model group rats, the cardiac weight index and cardiac-to-tibia ratio of the rats in each dose group of the traditional Chinese medicine prescription of this invention and the Tongxinluo capsule group were significantly decreased (P<0.05, P<0.01, P<0.001). However, there was no significant difference in cardiac weight index and cardiac-to-tibia ratio among the each dose group of the traditional Chinese medicine prescription of this invention and the Tongxinluo capsule group.

[0162] 3.3. Effects of the formulation of this invention on serum CK, CK-MB, cTnT, and MMP-9 levels in rats with myocardial infarction.

[0163] Figure 5A The bar chart shows the serum CK levels of rats in each group. Figure 5B The image shows a bar chart of serum CK-MB levels in each group of rats. Figure 6A The bar chart shows the serum cTnT content of rats in each group. Figure 6B The bar chart shows the serum MMP-9 levels in rats from each group. Compared to the sham-operated group, * p < 0.05 ** p < 0.01, *** p < 0.001; compared with the model group, # p < 0.05 ## p < 0.01, ### p < 0.001.

[0164] The results showed that, compared with the sham-operated group, the serum levels of CK, CK-MB, cTnT, and MMP-9 in the model group were significantly increased (P<0.05, P<0.01, P<0.001). Compared with the model group, the serum levels of CK, CK-MB, cTnT, and MMP-9 in all dosage groups of the herbal prescription of this invention and the Tongxinluo capsule group were significantly decreased (P<0.05, P<0.01, P<0.001).

[0165] 3.4. Effects of the formulation of this invention on cardiac tissue in rats with myocardial infarction

[0166] Figure 7 HE staining images (200×) of rat heart tissue from each group. A represents the sham-operated group, B the model group, C the low-dose prescription group, D the medium-dose prescription group, E the high-dose prescription group, and F the Tongxinluo capsule group.

[0167] Figure 7The results showed that the cardiac tissue structure of the model group rats was disordered, with fibrous connective tissue proliferation replacing cardiomyocytes, and inflammatory cell infiltration and neovascularization were also observed. Compared with the model group, the range of fibrous connective tissue proliferation and the number of inflammatory cells were reduced in a small number of samples from the Tongxinluo capsule group, and no obvious fibrosis was observed in a few samples. The range of fibrous connective tissue proliferation and the number of inflammatory cells were reduced in the cardiac tissues of rats in each dosage group of the traditional Chinese medicine prescription of this invention.

[0168] In summary, the cardiac weight index, myocardial enzymes, and myocardial injury indicators of rats with myocardial infarction were significantly reduced after intervention with the traditional Chinese medicine prescription of this invention, indicating that the prescription of this invention has a good therapeutic effect on myocardial infarction.

[0169] The above description is merely a specific embodiment of this disclosure, enabling those skilled in the art to understand or implement it. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this disclosure. Therefore, this disclosure is not to be limited to the embodiments described herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A traditional Chinese medicine composition for treating myocardial infarction, characterized in that, The traditional Chinese medicine composition includes the following components: Astragalus, Codonopsis, Ligusticum chuanxiong, Salvia miltiorrhiza, Lilium brownii, Lindera strychnifolia, Rehmannia glutinosa, Panax notoginseng, and Cyperus rotundus.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The traditional Chinese medicine composition comprises the following components in parts by weight: Astragalus membranaceus 22-26 parts, Codonopsis pilosula 18-23 parts, Ligusticum chuanxiong 7-11 parts, Salvia miltiorrhiza 13-17 parts, Lilium brownii 13-18 parts, Lindera strychnifolia 8-12 parts, Rehmannia glutinosa 18-23 parts, Panax notoginseng 10-14 parts, and Cyperus rotundus 7-12 parts.

3. The traditional Chinese medicine composition according to claim 1 or 2, characterized in that, The traditional Chinese medicine composition comprises the following components in parts by weight: Astragalus membranaceus 24 parts, Codonopsis pilosula 20 parts, Ligusticum chuanxiong 9 parts, Salvia miltiorrhiza 15 parts, Lilium brownii 15 parts, Lindera strychnifolia 10 parts, Rehmannia glutinosa 20 parts, Panax notoginseng 12 parts, and Cyperus rotundus 9 parts.

4. A pharmaceutical preparation for treating myocardial infarction, characterized in that, The pharmaceutical preparation contains the active ingredient of the traditional Chinese medicine composition as described in any one of claims 1-3.

5. The pharmaceutical preparation according to claim 4, characterized in that, The pharmaceutical preparation also includes excipients.

6. The pharmaceutical preparation according to claim 4 or 5, characterized in that, The dosage form of the pharmaceutical preparation is granules, tablets, capsules, or decoction.

7. A method for preparing a pharmaceutical formulation as described in any one of claims 4-6, characterized in that, The preparation method includes the following steps: (1) The traditional Chinese medicine composition according to any one of claims 1-3 is subjected to water extraction to obtain an aqueous extract; (2) The aqueous extract is prepared into the dosage form of the drug preparation.

8. The preparation method according to claim 7, characterized in that, The water extraction step includes: The herbal composition is decocted with water at least twice, each time for 1-4 hours. After each decoction, the mixture is filtered, and the filtrates are combined to obtain the aqueous extract.

9. The preparation method according to claim 7 or 8, characterized in that, In the water extraction step, the total mass of water is 18-35 times the mass of the traditional Chinese medicine composition.

10. The preparation method according to claim 8, characterized in that, In the water extraction step, the decoction is performed twice. In the first decoction, the mass of water is 10-20 times the mass of the Chinese herbal composition, and the decoction time is 2-3 hours. In the second decoction, the mass of water is 8-15 times the mass of the Chinese herbal composition, and the decoction time is 1-2 hours.

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