A traditional Chinese medicine composition for treating ischemic heart failure and a preparation method thereof

Abstract

The present application relates to a kind of traditional Chinese medicine composition for treating ischemic heart failure and preparation method thereof, belong to the field of traditional Chinese medicine pharmacy, the traditional Chinese medicine composition includes: cassia 5-15 parts, white atractylodes 5-15 parts, poria 5-15 parts, dried ginger 6-12 parts, acanthopanax 5-15 parts, water red flower seed 10-30 parts, verbena 3-9 parts, dogwood 6-12 parts, fried licorice 3-9 parts.The composition can significantly reduce N-terminal B-type natriuretic peptide level, significantly improve model rat heart tissue pathological changes, regulate the heart function of model rat, improve the systolic function of heart, and can reduce the expression of rat interleukin-1 beta, interleukin-6, tumor necrosis factor-alpha and other inflammatory factors, involve extracellular matrix receptor interaction, chemotactic factor signal pathway, relaxin signal pathway, cyclic adenosine monophosphate signal pathway, nuclear factor-kappa B signal pathway and multiple pathways, and is effective drug for treating ischemic heart failure.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of traditional Chinese medicine, and relates to a traditional Chinese medicine composition for treating heart failure, in particular to a traditional Chinese medicine composition for treating ischemic heart failure and a preparation method thereof. BACKGROUND

[0002] Heart failure is one of the major global public health problems. According to the global burden of disease research data, from 1990 to 2021, the number of patients with heart failure worldwide increased from 25.4 million to 55.5 million, showing a significant growth trend. The China Cardiovascular Health and Disease Report 2023 estimates that there are about 8.9 million heart failure patients in China. With the improvement of living standards, the etiological spectrum of heart failure in China has changed significantly, and ischemic heart disease has replaced rheumatic heart disease as the most common cause. Compared with non-ischemic heart failure, ischemic heart failure has a higher risk of cardiovascular events and death, and therefore becomes a clinical problem that needs to be urgently addressed.

[0003] At present, the main treatment strategy for ischemic heart disease is still centered on revascularization and drug therapy. Revascularization can improve myocardial perfusion and restore the contractile function of "stunned" or "hibernating" myocardium in some patients. However, the mechanism of myocardial ischemia is not only caused by coronary artery stenosis, but also closely related to many pathological factors such as coronary spasm, coronary microcirculation disorder, persistent activation of inflammatory response, abnormal platelet and coagulation function, and vascular endothelial dysfunction. A large number of studies have shown that chronic low-grade inflammation plays a key role in the progression of ischemic heart disease and heart failure, not only aggravating myocardial cell damage and fibrosis, but also interacting with overactivation of the neuroendocrine system to promote ventricular remodeling and continuous deterioration of cardiac function. In theory, early revascularization can improve resting blood flow and coronary blood flow reserve, rescue hibernating myocardium, and thus prevent or reverse ventricular remodeling; but in patients with ischemic heart failure, due to severe impairment of cardiac function, extensive and complex coronary artery lesions, the risk of revascularization-related surgery is significantly increased, and its benefit on long-term prognosis is still controversial. On the other hand, although drug therapy can delay disease progression by inhibiting neuroendocrine activation and improving hemodynamic status, patients still face a high risk of cardiovascular events even when the recommended drugs reach the maximum tolerated dose, and some patients have poor efficacy or poor tolerance. Therefore, the prevention and treatment of ischemic heart failure still faces severe challenges, and new intervention strategies or therapeutic drugs targeting key pathological links such as inflammation are urgently needed to improve disease outcomes from multiple target points. SUMMARY

[0004] Based on the above purpose, according to the advantages of traditional Chinese medicine in treating ischemic heart failure, such as accurate curative effect, less side effects and making up for the deficiency of western medicine treatment, a traditional Chinese medicine composition for treating ischemic heart failure and a preparation method thereof are provided to provide a new choice for clinical treatment drugs, and the specific technical scheme is as follows:

[0005] The first object of the present application is to provide a traditional Chinese medicine composition for treating ischemic heart failure, which is made of raw medicinal materials in the following proportions by weight: cassia twig 5-15 parts, white atractylodes 5-15 parts, poria 5-15 parts, dried ginger 6-12 parts, acanthopanax 5-15 parts, water red flower seed 10-30 parts, verbena 3-9 parts, dogwood 6-12 parts, and fried licorice 3-9 parts.

[0006] Further, the traditional Chinese medicine composition is made of raw medicinal materials in the following proportions by weight: cassia twig 10 parts, white atractylodes 10 parts, poria 10 parts, dried ginger 9 parts, acanthopanax 10 parts, water red flower seed 15 parts, verbena 6 parts, dogwood 9 parts, and fried licorice 6 parts.

[0007] The second object of the present application is to provide a preparation method of the traditional Chinese medicine composition for treating ischemic heart failure, which comprises the following steps:

[0008] Step 1: weigh each kind of traditional Chinese medicinal material according to the prescription proportion as raw material;

[0009] Step 2: according to the amount of the medicinal material taken in step 1, add 10-15 times the volume of water to pre-soak for 30-60 minutes;

[0010] Step 3: the soaked medicinal materials are decocted twice, and the filtrate is collected and combined;

[0011] Step 4: the combined decoction is concentrated to obtain a medicinal liquid paste;

[0012] Step 5: the obtained medicinal liquid is further processed to obtain the required dosage form preparation.

[0013] Further, the decoction method in step 3 is to first boil with strong fire and then change to medium fire for 60 minutes, filter to obtain the first decoction; after the decoction is finished, separate the medicinal liquid and the residue, add 8 times the amount of water to the residue, boil with strong fire and then change to medium fire for 30 minutes, filter to obtain the second decoction, and combine the filtered decoctions.

[0014] Further, the further processing in step 5 includes: using vacuum concentration, spray drying or freeze drying to obtain dry powder or extract powder.

[0015] Further, the dosage form of the traditional Chinese medicine composition for treating ischemic heart failure includes adding excipients, mixing, granulating or tabletting to prepare granules or tablets, and filling the obtained powder or granules into a hollow capsule to prepare a capsule; sterilizing the clear liquid and then dispensing to prepare an oral liquid; or adding a suitable binder to prepare water pills, honey pills or ointments.

[0016] Working principle of the present application:

[0017] Cassia twig: The product is the dried tender branch of Cinnamomum cassia Presl. It is pungent, sweet and warm, and belongs to the heart, lung and bladder meridians. It has the effects of sweating to relieve the exterior, warming and unblocking meridians, assisting yang and transforming qi, and reducing and descending qi. Modern pharmacological studies have shown that cassia twig contains volatile oil (cinnamyl aldehyde, cinnamic acid, cinnamyl alcohol), flavonoids (quercetin, kaempferol), polysaccharides, coumarin and other components. Cinnamyl aldehyde can dilate the coronary artery, reduce peripheral resistance, improve myocardial microcirculation, inhibit platelet aggregation and reduce blood viscosity, and has antioxidant and anti-oxidative effects to reduce the generation of oxygen free radicals and reduce ventricular remodeling.

[0018] White atractylodes: The product is the dried rhizome of Atractylodes macrocephala Koidz. It is bitter, sweet and warm, and belongs to the spleen and stomach meridians. It has the effects of invigorating the spleen and tonifying qi, drying dampness and promoting water, stopping sweating and securing the fetus. Modern pharmacological studies have shown that white atractylodes mainly contains sesquiterpene lactones (atractylenolide, atractylenolide), polysaccharides, flavonoids and other components. White atractylodes polysaccharide can enhance the antioxidant capacity of myocardial cells; atractylenolide II and III can reduce inflammatory cell infiltration in myocardial tissue after myocardial infarction, inhibit platelet activation and myocardial fibrosis.

[0019] Poria cocos: The product is the dried sclerotium of Poria cocos (Schw.) Wolf. It is sweet and mild, and belongs to the heart, lung, spleen and kidney meridians. It has the effects of promoting water and removing dampness, invigorating the spleen and calming the heart. Modern pharmacological studies have shown that Poria cocos contains triterpenes (pachymic acid, pachymic acid), pachyman, sterols and other components. Pachyman has diuretic, immunoregulatory and antioxidant effects, triterpenes can block the renin-angiotensin-aldosterone system, reduce angiotensin II levels, reduce ventricular remodeling, and have anti-inflammatory effects, and have protective effects on ischemic heart failure.

[0020] Dried ginger: The product is the dried rhizome of Zingiber officinale Rosc. It is pungent and warm, and belongs to the spleen, stomach, kidney, heart and lung channels. It has the effects of warming middle energizer, dispelling cold, restoring yang and unblocking collaterals, and warming lung and resolving fluid. Modern pharmacological studies have shown that dried ginger mainly contains gingerols (6-gingerol, 8-gingerol, 10-gingerol), gingerenol, flavonoids, volatile oils and other ingredients. 6-gingerol can improve myocardial energy metabolism by increasing sarcoplasmic reticulum / endoplasmic reticulum calcium ion ATPase activity, inhibit the nuclear factor-kappa B pathway, and reduce myocardial inflammatory response. In ischemic heart failure, the active ingredients of dried ginger can also reduce myocardial oxidative stress and improve cardiac function.

[0021] Eleutherococcus senticosus: The product is the dried root and rhizome or stem of Acanthopanax senticosus (Rupr. et Maxim.) Harms of the Araliaceae family. It is pungent and slightly bitter, and warm. It belongs to the spleen, kidney and heart channels. It has the effects of benefiting qi and invigorating the spleen, and tonifying the kidney and tranquilizing the mind. Modern pharmacological studies have shown that eleutherococcus contains flavonoids, organic acids, lignans, coumarins and other ingredients. Among them, eleutheroside B can reduce myocardial necrotic apoptosis and improve myocardial damage through the nuclear factor-erythroid factor 2-related factor 2 / heme oxygenase-1 signaling pathway. Clinical studies have shown that eleutherococcus can improve the left ventricular diastolic function of patients with angina pectoris, and has the effects of dilating coronary arteries, increasing coronary blood flow, slowing heart rate and inhibiting myocardial contractility.

[0022] Verbena officinalis: The product is the dried aboveground part of Verbena officinalis L. of the Verbena officinalis L. It is bitter and cool, and belongs to the liver and spleen channels. It has the effects of activating blood and resolving stasis, detoxifying, promoting water, reducing yellow and stopping malaria. Modern pharmacological studies have shown that verbena mainly contains iridoid glycosides (verbascoside, hibiscus glycoside), flavonoids (luteolin, apigenin), phenethyl glycosides and other ingredients. Verbascoside can inhibit platelet activation and aggregation, improve coronary microcirculation, and reduce the degree of myocardial ischemia; ursolic acid can inhibit the transforming growth factor-beta signaling pathway, reduce myocardial collagen deposition, and delay the progression of myocardial fibrosis.

[0023] Water red seed: The product is the dried mature fruit of Polygonum orientale L. of the Polygonaceae family. It is salty and slightly cold, and belongs to the liver and stomach channels. It has the effects of dispersing blood, resolving symptoms, resolving accumulation and relieving pain, and promoting water and reducing swelling. Modern pharmacological studies have shown that water red seed contains flavonoids (such as taxifolin, quercetin, iridoid glycosides, etc.), amides (p-coumaroyl tyramine, p-feruloyl tyramine, etc.), alkaloids and other ingredients. Quercetin and taxifolin can reduce lipid peroxidation, improve myocardial ischemia and hypoxia, and inhibit myocardial cell apoptosis.

[0024] Cornus officinalis Sieb. et Zucc. ) of Cornaceae. Sour, astringent, and slightly warm. It belongs to the liver and kidney channels. It has the effects of tonifying liver and kidney and astringing and consolidating. Modern pharmacological studies have shown that Cornus officinalis contains iridoid glycosides (loganin, loganic acid), organic acids, flavonoids, polysaccharides and other components. Loganic acid can scavenge oxygen free radicals, restore mitochondrial respiratory function to improve cardiac remodeling after myocardial infarction, and delay the development of heart failure.

[0025] Glycyrrhiza uralensis Fisch., Glycyrrhiza inflata Bat. or Glycyrrhiza glabra L. of Leguminosae. It is a processed product of Glycyrrhiza, taking Glycyrrhiza slices, and frying according to the honey frying method (frying to yellow to dark yellow, taking out when not sticky, and airing). Sweet, and flat. It belongs to the heart, lung, spleen, and stomach channels. It has the effects of tonifying the spleen and stomach, and benefiting qi and restoring pulse. Modern pharmacological studies have shown that honey-fried Glycyrrhiza contains triterpenoid saponins (glycyrrhizic acid, 18β-glycyrrhizinic acid), flavonoids (liquiritin, isoliquiritigenin), polysaccharides and other components. Glycyrrhizic acid and its metabolites can regulate potassium channel current, improve myocardial electrical activity, and reduce the incidence of arrhythmia. Glycyrrhizinic acid improves heart function by up-regulating the nuclear factor-hemoglobin oxygenase 2-related factor 2 / hemoglobin oxygenase 1 signaling pathway to inhibit oxidative stress, apoptosis and regulate mitochondrial function.

[0026] Compatibility principle: Ginger and Ramulus Cinnamomi are the monarch drugs, Atractylodes Macrocephala and Poria Cocos are the minister drugs, Acanthopanax senticosus, Verbena Officinalis, Fructus Rubi, and Cornus officinalis are the assistant drugs, and honey-fried Glycyrrhiza is the minister drug. The nine components cooperate with each other, are warm but not dry, beneficial but not harmful, and have both dispersing and astringing effects, and the composition ratio is based on the nature, efficacy and synergistic effect of each component, and is obtained through multiple rounds of orthogonal experiments: the dosage ratio of ginger and Ramulus Cinnamomi is controlled at 1:1-1.5, which can avoid the dry heat reaction caused by excessive warming of yang, and maximize the effect of warming and unblocking heart yang; the dosage ratio of Acanthopanax senticosus and Cornus officinalis is 1:1, which takes into account the effects of tonifying qi and tonifying liver and kidney, and avoids the greasy and stomach-stifling caused by single tonification; the dosage ratio of Fructus Rubi and Verbena Officinalis is 2:1, which enhances the effects of promoting blood circulation and diuresis, and avoids the risk of bleeding caused by excessive blood circulation. The above ratio is verified by clinical pre-experiments, which is the key to achieving 'warming yang without dryness, promoting blood circulation without damaging the normal, and diuresis without consuming qi'.

[0027] The dried ginger and the ramulus cinnamomi in the prescription are used as the monarch drug, the ramulus cinnamomi tastes acrid and sweet, is warm in nature, and is attributed to the heart, lung and bladder meridians, can directly supplement heart yang, and can also warm and unblock meridians, guide other drugs to the heart meridian, and enhance the targeting effect of the whole prescription on the cardiovascular system, meanwhile, the dried ginger can warm and transport the spleen yang, and provide support for the heart yang; meanwhile, the heat of the dried ginger can enhance the effect of the ramulus cinnamomi on warming the heart yang, and improve the symptoms of heart yang deficiency, such as fear of cold, cold hands and feet, pale tongue and white coating, etc.

[0028] The atractylodes and the poria are used as the ministerial drug: the two are a classic combination of 'invigorating the spleen and promoting diuresis' (derived from the Lingui Zhugan Decoction); the atractylodes tastes sweet and bitter, is warm in nature, and is attributed to the spleen and stomach meridians, can invigorate the spleen and boost qi, and dry dampness and promote diuresis - by supplementing the spleen qi and invigorating the spleen and stomach, the water and dampness retention caused by the spleen yang deficiency is improved (water and dampness overflow easily to the heart and lung, and aggravate palpitation and shortness of breath, or overflow to the skin to cause edema), and provide the basis for 'promoting diuresis' by'spleen transportation'; the poria tastes sweet and bland, is neutral in nature, and is attributed to the heart, lung, spleen and kidney meridians, can promote diuresis and remove dampness, and invigorate the spleen and calm the heart - on the one hand, it helps the atractylodes to enhance the diuretic effect, and remove excess water and dampness in the body (reduce the burden on the cardiovascular system); on the other hand, its 'calming the heart' effect can directly improve palpitation and insomnia, and form a synergistic effect of 'warming yang and calming the heart' with the 'warming heart yang' of the ramulus cinnamomi.

[0029] The accompanying drugs are the acanthopanax, the cornus, the water red flower seed and the verbena - tonifying qi and kidney, promoting blood circulation and diuresis, and taking into account 'complementary symptoms' and 'balance', the accompanying drugs in the prescription are 'assistant drugs', which assist the monarch and ministerial drugs to enhance the curative effect.

[0030] Compared with the prior art, the present application has the following advantages:

[0031] Firstly, the traditional Chinese medicine composition in the present application does not have a complete corresponding formula, and the prescription idea is similar to the classic prescription Lingui Zhugan Decoction in terms of warming yang and transforming dampness, and invigorating the spleen and removing dampness, but the present application has the following advantages:

[0032] 1. The addition of the acanthopanax breaks through the limitation of the Lingui Zhugan Decoction in only warming and supplementing the middle yang, can consider supplementing kidney qi while warming the spleen yang, so that the prescription is not only suitable for water and dampness retention caused by simple middle yang deficiency, but also makes the 'fire' of warming yang rooted in the lower jiao and the conception vessel, so that the qi transformation of the triple energizer has a source.

[0033] 2. The combination of the verbena and the water red flower seed adds the characteristics of promoting blood circulation and diuresis to the prescription, and the two can break down stasis while promoting diuresis when used together, solve the shortcoming of the Lingui Zhugan Decoction in only promoting diuresis but not transforming stasis, and are more suitable for improving water and sodium retention in patients with ischemic heart failure.

[0034] 3. The addition of dogwood achieves the balance of tonifying and astringing and eliminating evil without damaging the healthy, on the one hand, dogwood assists acanthopanax to enhance the kidney tonifying power, on the other hand, it can restrict the dissipation of water and blood of verbena and water red flower seed, avoiding the hidden danger of deficiency of qi and yin caused by simple water diuresis in long-term use of the classic Lingui Zhugan Decoction, reflecting the organic extension and clinical upgrading of modern Chinese medicine to the classic path.

[0035] 4. Animal tests prove that the traditional Chinese medicine composition can significantly reduce the N-terminal B-type natriuretic peptide level, can significantly improve the pathological changes of heart tissue of the model rats, can regulate the heart function of the rats, especially can improve the systolic function of the heart, and can reduce the expression of inflammatory factors such as interleukin-1β, interleukin-6 and tumor necrosis factor alpha, and the mechanism thereof involves multiple pathways such as extracellular matrix receptor interaction, chemokine signaling pathway, relaxin signaling pathway, cyclic adenosine monophosphate signaling pathway, nuclear factor-kappa B signaling pathway, fatty acid metabolism and Toll-like receptor signaling pathway, and is an effective drug for treating ischemic heart failure.

[0036] 5. When the traditional Chinese medicine composition in the application is removed, the regulation effect of the traditional Chinese medicine composition on heart failure markers and inflammatory factors is significantly weakened. BRIEF DESCRIPTION OF DRAWINGS

[0037] Figure 1 The ultrasound cardiogram detection results of the rats are shown in the table, wherein: (A) is a representative image of the ultrasound cardiogram of each group; (B) is the change of left ventricular ejection fraction (LVEF) of each group; (C) is the change of left ventricular shortening fraction (LVFS) of each group. Compared with the blank control (sham operation group), *P<0.05; **P<0.01; ***P<0.001; compared with the model group, #P<0.05; ##P<0.01; ###P<0.001.

[0038] Figure 2 The serum factor detection results of the rats are shown in the table, wherein: (A) is the N-terminal B-type natriuretic peptide level of each group; (B) is the interleukin-1β level of each group; (C) is the interleukin-6 level of each group; (D) is the tumor necrosis factor-α level of each group. Compared with the blank control group, #P<0.05; ##P<0.01; ###P<0.001; compared with the model group, *P<0.05; **P<0.01; ***P<0.001.

[0039] Figure 3 The heart tissue pathological section staining of the rats is shown in the table, wherein: (A) is the HE staining result of each group, (B) is the Masson staining result of each group.

[0040] Figure 4Figure 8 is a volcano plot of differentially expressed genes between the model group and the traditional Chinese medicine group for the rat heart transcriptome results; wherein: (A) is a volcano plot of differentially expressed genes between the model group and the traditional Chinese medicine group; (B) is a pathway enrichment analysis of differentially expressed genes between the model group and the traditional Chinese medicine group.

[0041] Figure 5 Figure 9 is a rat liver and kidney function result; wherein: (A) is the alanine aminotransferase level of each group, (B) is the aspartate aminotransferase level of each group; (C) is the blood creatinine level of each group; (D) is the blood urea level of each group.

[0042] Figure 6 Figure 10 is the regulatory effect of the traditional Chinese medicine composition without dried ginger, acanthopanax, water red flower seed, verbena and dogwood on ischemic heart failure; wherein: (A) is the N-terminal B-type natriuretic peptide level of each group; (B) is the interleukin-1β level of each group; (C) is the interleukin-6 level of each group; (D) is the tumor necrosis factor-α level of each group. Compared with the blank control (sham operation group), *P<0.05; **P<0.01; ***P<0.001; compared with the model group, #P<0.05; ##P<0.01; ###P<0.001; compared with the composition without dried ginger, acanthopanax, water red flower seed, verbena and dogwood, +P<0.05; ++P<0.01; +++P<0.001. DETAILED DESCRIPTION

[0043] The application will be described in further detail below with specific embodiments in conjunction with the accompanying drawings.

[0044] Example 1: A preparation method of a traditional Chinese medicine composition for treating ischemic heart failure, comprising the following steps: 10 parts of cassia twig, 10 parts of white atractylodes, 10 parts of poria, 9 parts of dried ginger, 10 parts of acanthopanax, 15 parts of water red flower seed, 6 parts of verbena, 9 parts of dogwood, and 6 parts of fried licorice are soaked in 700 ml of water for 30 minutes; after boiling with a strong fire, the fire is changed to a weak fire for 60 minutes, and the first decoction is filtered; after the decoction ends, the medicinal liquid and the medicinal residue are separated, 8 times the amount of water is added to the medicinal residue, and after boiling with a strong fire, the fire is changed to a weak fire for 30 minutes, and the second decoction is filtered; the filtrates are combined. After further adding conventional granule adjuvants and sterilizing, granules are prepared according to the conventional method.

[0045] Example 2: A preparation method of a traditional Chinese medicine composition for treating ischemic heart failure, comprising the following steps: 5 parts of cassia twig, 5 parts of white atractylodes, 5 parts of poria, 6 parts of dried ginger, 5 parts of acanthopanax, 10 parts of water red flower seed, 3 parts of verbena, 6 parts of dogwood, and 3 parts of fried licorice are soaked in 700 ml of water for 30 minutes; after boiling with a strong fire, the fire is changed to a weak fire for 60 minutes, and the first decoction is filtered; after the decoction ends, the medicinal liquid and the medicinal residue are separated, 10 times the amount of water is added to the medicinal residue, and after boiling with a strong fire, the fire is changed to a weak fire for 30 minutes, and the second decoction is filtered; the filtrates are combined. After further adding conventional granule adjuvants and sterilizing, granules are prepared according to the conventional method.

[0046] Example 3: A preparation method of a traditional Chinese medicine composition for treating ischemic heart failure, comprising the following steps: 15 parts of guizhi, 15 parts of baizhu, 15 parts of fuling, 12 parts of ganjiang, 15 parts of cigou, 30 parts of shuihonghuazi, 9 parts of maibangcao, 12 parts of shanzhu, and 9 parts of zhigancao are soaked in 1000 ml of water for 30 minutes; after boiling with strong fire, the fire is changed to weak fire for 60 minutes, and the first decoction is obtained by filtration; after the end of decoction, the medicinal liquid and the residue are separated, 15 times the amount of water is added to the residue, and after boiling with strong fire, the fire is changed to weak fire for 30 minutes, and the second decoction is obtained by filtration; the fourth decoction is obtained by repeating the decoction and combining the filtrates. After sterilization by further adding conventional granule adjuvants, granules are prepared by conventional methods.

[0047] Test Example 1: Animal pharmacology experiment (using the pharmaceutical composition of Example 1)

[0048] 1. Materials and methods

[0049] Animal research experiment using the traditional Chinese medicine composition of Example 1

[0050] Eighty-five specific pathogen-free male SD rats weighing 200±20 g were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. The rats were given free access to standard synthetic feed and pure water, and the feeding conditions were temperature (24℃±2℃), humidity (45%-50%), and 12 hours of light per day. A myocardial infarction model was established in rats by ligating the left anterior descending branch by skilled personnel. Briefly, the rats were anesthetized using isoflurane ventilation, fixed in a supine position on a dissecting table, shaved, prepared for skin, and disinfected. A tube was connected to an animal ventilator (tidal volume 3 ml / 100 g body weight, breathing frequency 60 times / minute). The ribs were separated to expose the heart, and a surgical silk thread was used to ligate 2-3 mm below the left atrial appendage. Ligation success was confirmed by electrocardiogram examination (e.g., ST segment elevation) and immediate pallor of the ligation area. In the sham operation group, only the thread was passed, and no arterial ligation was performed. After the myocardial infarction model was successfully prepared, the rats were normally fed for 1 week for echocardiography detection. A heart ultrasound LVEF<50% was considered a successful ischemic heart failure model. The model rats were randomly divided into 6 groups using a random number table method, in order of sham operation group, model group, traditional Chinese medicine composition low-dose group (3.75 g / kg / d), traditional Chinese medicine composition medium-dose group (7.5 g / kg / d), traditional Chinese medicine composition high-dose group (15 g / kg / d), and enalapril group (0.45 mg / kg / d), and continuous administration was performed for 28 days. The animal drug dose was equivalent to the human body surface area and body weight. The rats in the Sham group and Model group were given the same amount of phosphate buffered saline.

[0051] 2. Echocardiography detection

[0052] Anesthetized rats were used by two-dimensional M-mode and B-mode echocardiography to detect animal heart function by technicians who were unaware of the grouping, and the results were as followsFigure 1 As shown in Table 1, the model group showed a significant decrease in LVEF and FS compared with the sham group, indicating that the model group successfully constructed a heart function injury model.

[0053] Figure 1 A (echocardiography): The myocardial movement of the sham group showed regular waveforms, and the ventricular wall contraction and diastole were coordinated. The myocardial waveforms of the model group were disordered, and the ventricular wall movement was abnormal, indicating impaired heart function. The myocardial waveforms of the different dose treatment groups and the enalapril group (positive control group, angiotensin converting enzyme inhibitor) gradually improved compared with the model group.

[0054] Figure 1 B (left ventricular ejection fraction, LVEF): The LVEF value of the sham group was relatively high (about 80% or more); the LVEF of the model group was significantly reduced (P<0.01, indicating a very significant difference compared with the sham group); the LVEF of the medium and high dose groups was significantly higher than that of the model group (#, ## indicating significant or very significant differences compared with the model group), indicating that the treatment could improve the left ventricular ejection function.

[0055] Figure 1 C (fractional shortening, FS): The FS value of the sham group was relatively high (about 50% or more); the FS of the model group was significantly reduced (P<0.01, indicating a very significant difference compared with the sham group); the FS of the medium and high dose groups was significantly higher than that of the model group (#, ## indicating significant or very significant differences compared with the model group).

[0056] In summary, compared with the blank control group, the heart structure of the model group changed, and the left ventricular ejection fraction and fractional shortening decreased. After treatment with different doses of traditional Chinese medicine, the left ventricular ejection fraction and fractional shortening significantly increased (P<0.05). The above results show that the model group successfully constructed a heart function injury model, and treatment with different doses of traditional Chinese medicine composition (especially medium and high doses) can improve the movement function of myocardium, increase the left ventricular ejection fraction (LVEF) and fractional shortening (FS), and thus improve the heart function and increase the contraction function of the heart, further indicating that the traditional Chinese medicine composition plays a protective role on the heart and can provide experimental basis for the treatment or intervention of related heart diseases.

[0057] 3. Serological detection

[0058] After the rats were anesthetized, blood was collected from the abdominal aorta, and after standing at room temperature for 1 hour, it was centrifuged at 3000 rpm for 10 minutes in a 4°C centrifuge. The levels of N-terminal B-type natriuretic peptide, interleukin-1β, interleukin-6, and tumor necrosis factor-α were detected by enzyme-linked immunosorbent assay according to the instructions, and the results are shown in Table 2. Figure 2

[0059] Figure 2 ​​​A (N-terminal pro-B-type natriuretic peptide, NT-proBNP level): The NT-proBNP level of the Sham group (sham operation group) was low; the NT-proBNP level of the model group was significantly increased (## indicates a very significant difference compared with the Sham group); the NT-proBNP levels of the low-dose group, the medium-dose group, the high-dose group and the enalapril (positive control group, angiotensin converting enzyme inhibitor) group were significantly lower than those of the model group, and the medium-dose, high-dose and enalapril groups were more significantly reduced (#, ##, ### indicates a significant or very significant difference compared with the model group).

[0060] Figure 2 B (interleukin-1β, IL-1β level): The IL-1β level of the Sham group was low; the IL-1β level of the model group was significantly increased (## indicates a very significant difference compared with the Sham group); the IL-1β levels of the different dose groups and the enalapril group were significantly lower than those of the model group (#, ##, ### indicates a significant or very significant difference compared with the model group).

[0061] Figure 2 C (interleukin-6, IL-6 level): The IL-6 level of the Sham group was low; the IL-6 level of the model group was significantly increased (## indicates a very significant difference compared with the Sham group); the IL-6 levels of the different dose groups and the enalapril group were lower than those of the model group, and the medium-dose, high-dose and enalapril groups were more significantly reduced (#, ##, ### indicates a significant or very significant difference compared with the model group).

[0062] Figure 2 D (tumor necrosis factor-α, TNF-α level): The TNF-α level of the Sham group was low; the TNF-α level of the model group was significantly increased (## indicates a very significant difference compared with the Sham group); the IL-6 levels of the different dose groups and the enalapril group were lower than those of the model group, and the medium-dose, high-dose and enalapril groups were more significantly reduced (#, ##, ### indicates a significant or very significant difference compared with the model group).

[0063] ​​​​The above results show that, compared with the blank control group, the heart failure markers (N-terminal B-type natriuretic peptide) and inflammatory indicators (interleukin-1 beta, interleukin-6, tumor necrosis factor-alpha) of the model group are significantly increased, and different doses of treatment (especially medium and high doses) and ACEI treatment can reduce the NT-proBNP level, improve the heart function related indicators, and significantly inhibit the expression of inflammatory factors IL-1 beta, IL-6 and TNF-alpha, reduce the inflammatory response, indicating that the traditional Chinese medicine composition has an improvement effect on heart failure, and can reduce the heart inflammatory indicators, and plays a protective role on the heart through anti-inflammatory mechanism, providing an experimental basis for the treatment or intervention of related heart diseases.

[0064] 4. Heart tissue pathological section staining

[0065] The rat heart tissue was fixed in 4% paraformaldehyde overnight. The fixed tissue was embedded in paraffin to make 4-micron sections. Hematoxylin-eosin and Masson staining were performed according to the instructions, and the results are shown in Figure 3 As shown in the results, compared with the blank control group, Figure 3 A HE staining result shows that the myocardial cell structure of the model group is disordered and arranged loosely, the cell nucleus is pyknosis or fragmentation, and obvious immune infiltration appears. After treatment with traditional Chinese medicine, the immune infiltration is reduced, and the cell structure and arrangement integrity are reserved. Compared with the blank control group, Figure 3 B Masson staining shows that the myocardial cells of the model group have obvious blue collagen fiber distribution, and the traditional Chinese medicine treatment can reduce collagen deposition. This shows that the traditional Chinese medicine composition has a protective effect on the heart tissue.

[0066] The results show that the model group successfully constructs a model of myocardial injury (possibly accompanied by myocardial fibrosis). The medium and high dose treatments can improve the pathological morphology of myocardial tissue and reduce the degree of myocardial fibrosis, and play a protective role on the heart tissue, providing an experimental basis at the histological level for the treatment or intervention of related heart diseases.

[0067] 5. Transcriptional analysis of the potential mechanism of the traditional Chinese medicine composition

[0068] The left ventricular myocardial tissue of experimental animals is taken, total RNA is extracted by a conventional method, and the RNA integrity is detected by agarose gel electrophoresis, and the RNA purity and concentration are detected by ultraviolet spectrophotometry or a biological analyzer. After the qualified RNA sample is subjected to mRNA enrichment or rRNA removal, cDNA is synthesized by fragmentation and reverse transcription, and a sequencing library is obtained after end repair, A tailing and adapter ligation. After the library is qualified by quality detection, double-end sequencing is performed by using a high-throughput sequencing platform. The original sequencing data are subjected to quality control, and low-quality reads and adapter sequences are removed to obtain high-quality clean reads. The clean reads are aligned to a reference genome or a transcript database, and the expression of genes or transcripts is calculated. According to sample grouping, the gene expression in different groups is statistically analyzed, and differentially expressed genes are screened. The standard for differential screening can be that the absolute value of the logarithmic conversion fold change is > 0.25 and the significance level P value is < 0.05. The differentially expressed genes are subjected to pathway enrichment analysis to explore their possible biological functions and signal pathways, and the results are shown in Figure 4 115 significantly changed differential genes are identified in the model group and the traditional Chinese medicine group, and the differential genes are subjected to pathway enrichment, which is mainly related to extracellular matrix receptor interaction, chemokine signaling pathway, relaxin signaling pathway, cyclic AMP signaling pathway, nuclear factor-kappa B signaling pathway, fatty acid metabolism, Toll-like receptor signaling pathway and the like. This result shows that the traditional Chinese medicine composition of the present application has a protective effect on heart tissue and has a multi-target and multi-pathway mechanism of action.

[0069] 6. Safety indicators

[0070] After the experimental animals in each group are subjected to corresponding treatment, blood samples are collected. Then, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum creatinine (Scr) and urea (UREA) in the blood are detected by using biochemical detection methods.

[0071] The results are shown in the results of liver function and kidney function. Figure 5 During the entire experiment, compared with the normal group, no differences in the liver function indicators alanine aminotransferase and aspartate aminotransferase and no differences in the kidney function indicators creatinine and urea are found in the traditional Chinese medicine composition groups of different doses, which shows that the traditional Chinese medicine composition of the present application is safe.

[0072] 7. Regulation effect of the traditional Chinese medicine composition of the present application on ischemic heart failure

[0073] The results are shown in Figure 6In all four indicators, the expression of the model group was significantly higher than that of the sham operation group, indicating that the experimental modeling was successful, and the animal model had obvious inflammatory response and heart failure related pathological changes. And the Lingguizhugan Decoction (not containing the dried ginger, acanthopanax, water red flower seed, verbena and dogwood of the present application) and the positive Chinese medicine can significantly reduce the level of the index: compared with the model group, the expression of NT-proBNP, IL-1β, IL-6 and TNF-α in all dosing groups of Lingguizhugan Decoction (3.2g / kg, 6.4g / kg) and positive Chinese medicine (7.5g / kg, 15g / kg) was significantly reduced, proving that both drugs can effectively improve the inflammatory response and heart failure state. However, it can be seen that the intervention effect of Lingguizhugan Decoction is weaker than that of the traditional Chinese medicine composition of the present application, such as the regulation of heart failure markers (N-terminal B-type natriuretic peptide) and inflammatory factors (interleukin-1β, interleukin-6, tumor necrosis factor-α levels) is significantly weakened (see Figure 6 ), indicating that the efficacy of the traditional Chinese medicine composition of the present application is closely related to the type of each medicinal ingredient in the prescription.

[0074] The dried ginger in the prescription is warm in nature and pungent in taste, and enters the heart, lung, spleen and stomach meridians, and can enhance the effect of warming and unblocking of guizhi, and is aimed at the core pathogenesis of 'heart yang deficiency' of patients with ischemic heart failure, and can relieve symptoms such as aversion to cold, cold limbs, palpitation, etc., and can significantly improve the effect of warming yang in cooperation with guizhi; and acanthopanax is warm in nature, pungent and slightly bitter in taste, and enters the spleen, kidney and heart meridians, and has the effects of tonifying qi and spleen, tonifying kidney and calming the mind, and promoting blood circulation to remove blood stasis, and can improve the symptoms of fatigue and shortness of breath caused by myocardial ischemia, and also contains acanthopanax saponin which can be used to protect myocardial cells and reduce ischemia-reperfusion injury; water red flower seed is cold in nature and salty in taste, and enters the liver and stomach meridians, and can promote blood circulation and remove stasis, and tonify the spleen and drain dampness, and is aimed at the symptoms of blood stasis and edema often accompanied by ischemic heart failure, and has the effect of promoting water and removing edema in cooperation with poria and atractylodes in the prescription, and can also improve myocardial microcirculation; the addition of verbena can enhance the blood circulation and remove blood stasis of the whole prescription, promote the dispersion of blood stasis and improve the myocardial ischemia state through its effect of promoting blood circulation and removing blood stasis and promoting water and removing edema; and the use of dogwood is to enhance the body's tolerance and reduce adverse reactions of drugs.

[0075] In summary, the traditional Chinese medicine composition of the present application shows good effects in improving heart function, reducing myocardial injury and inflammation, regulating myocardial fibrosis, etc., and has a multi-target and multi-pathway mechanism, which provides strong experimental evidence for its clinical treatment of heart diseases (such as ischemic heart failure, myocardial injury related diseases, etc.). Future clinical studies can be further carried out to verify its efficacy and safety in the human body, and its molecular mechanism of action can be further explored, providing more support for the modernization of traditional Chinese medicine and the precise treatment of heart diseases. In addition, the combination of the traditional Chinese medicine composition with existing clinical drugs (such as ACEI, etc.) can be further explored to play a synergistic role and improve the treatment effect.

Claims

1. A traditional Chinese medicine composition for treating ischemic heart failure, characterized in that, The medicine is made from the following raw materials in parts by weight: 5-15 parts cinnamon twig, 5-15 parts atractylodes macrocephala, 5-15 parts poria cocos, 6-12 parts dried ginger, 5-15 parts eleutherococcus senticosus, 10-30 parts water lily seeds, 3-9 parts verbena, 6-12 parts cornus officinalis, and 3-9 parts prepared licorice root. In this traditional Chinese medicine composition, cinnamon twig and dried ginger are the principal herbs, atractylodes macrocephala and poria cocos are the assistant herbs, eleutherococcus senticosus, verbena, water lily seeds, and cornus officinalis are the adjuvant herbs, and prepared licorice root is the guiding herb.

2. The traditional Chinese medicine composition for treating ischemic heart failure according to claim 1, characterized in that, It is made from the following raw materials in parts by weight: 10 parts cinnamon twig, 10 parts atractylodes macrocephala, 10 parts poria cocos, 9 parts dried ginger, 10 parts eleutherococcus senticosus, 15 parts water lily seeds, 6 parts verbena, 9 parts cornus officinalis, and 6 parts prepared licorice root.

3. A method for preparing a traditional Chinese medicine composition for treating ischemic heart failure according to any one of claims 1-2, characterized in that, Includes the following steps: Step 1: Weigh out each Chinese medicinal herb as raw material according to the prescription ratio; Step 2: Soak the herbs in 10-15 times their volume of water for 30-60 minutes, based on the amount of herbs taken in Step 1. Step 3: Boil the soaked herbs twice, filter the liquids separately, and combine them. Step 4: Concentrate the combined decoction to obtain a medicinal paste; Step 5: Further process the obtained liquid ointment to prepare the desired dosage form.

4. The method for preparing a traditional Chinese medicine composition for treating ischemic heart failure according to claim 3, characterized in that, The decoction method in step 3 is to first bring it to a boil over high heat, then simmer it over low heat for 60 minutes, and filter it to obtain the first decoction. After the decoction is finished, separate the decoction from the dregs. Add 8 times the amount of water to the dregs, bring it to a boil over high heat, then simmer it over low heat for 30 minutes, and filter it to obtain the second decoction. Combine the filtered decoctions.

5. The method for preparing a traditional Chinese medicine composition for treating ischemic heart failure according to claim 3, characterized in that, The further processing in step 5 includes: obtaining dry powder or extract powder by vacuum concentration, spray drying or freeze drying.

6. The method for preparing a traditional Chinese medicine composition for treating ischemic heart failure according to claim 3, characterized in that, The preparation of the required dosage form includes: adding excipients and mixing, granulating or compressing to make granules or tablets, or filling powder or granules into empty capsules to prepare capsules, or sterilizing and dispensing clarified drug liquid to prepare oral liquid, or adding suitable binders to make water pills, honey pills or ointments.

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