A traditional Chinese medicine composition for treating chronic heart failure

Abstract

The application discloses a traditional Chinese medicine composition for treating chronic heart failure, which is made from the following raw materials: Huangqi, Hei Shunpian, Dangshen, Lujiao, Dangui, Chishao, Tinglizi, Chao Baishu, Zexie, Zhishi, Fuling, Honghua and Guizhi. The traditional Chinese medicine composition has the functions of benefiting qi and warming yang, removing blood stasis and promoting water excretion, and is used as a ministerial medicinal. Zexie can promote water excretion, remove dampness, discharge heat and transform turbid and reduce fat; Zhishi can break qi, eliminate accumulation, transform phlegm, open chest and qi, and relax qi stagnation and blood stasis; the two medicines can regulate qi and blood, dredge lung qi, and disperse phlegm, water retention and accumulation, so as to achieve the effect of heart-lung coordination; Fuling can promote water excretion, remove dampness, strengthen the spleen and calm the heart, so as to enhance the effect of Huangqi on strengthening the spleen and promoting water excretion. Honghua can activate blood, dredge channels, disperse blood stasis and relieve pain; Guizhi can warm and dredge channels, assist yang and qi, and balance and descend, so as to assist Hei Shunpian in warming and tonifying kidney yang, dredging yang and qi; at the same time, the traditional Chinese medicine composition can enter the heart meridian. The traditional Chinese medicine composition has the functions of benefiting qi and warming yang, removing blood stasis and promoting water excretion.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine, specifically relating to a traditional Chinese medicine composition for treating chronic heart failure. Background Technology

[0002] Chronic heart failure (CHF) is a complex clinical syndrome caused by structural or functional abnormalities of the heart, leading to insufficient ventricular ejection and / or filling function. The estimated global prevalence of heart failure exceeds 37.7 million people, and the incidence is continuously rising. Its 5-year survival rate is similar to that of malignant tumors, making it one of the most important cardiovascular diseases of the 21st century. How to effectively treat heart failure has become a crucial issue worldwide. With in-depth research into its mechanisms, CHF is believed to be a syndrome involving multiple mechanisms, primarily the activation of the renin-angiotensin-aldosterone system (RAAS) and neuroendocrine disorders.

[0003] Traditional Chinese medicine (TCM) classifies chronic heart failure as a disease encompassing "phlegm retention," "edema," "asthma," and "palpitation." It is a condition of deficiency in the root and excess in the branch; the root deficiency refers to a deficiency of heart qi and yang, possibly accompanied by a deficiency of heart yin, while the branch excess involves blood stasis, fluid retention, and phlegm. The disease is located in the heart but is closely related to the lungs, spleen, and kidneys. Deficiency of heart qi and yang is a crucial underlying factor in the pathogenesis of heart failure, blood stasis is a central pathological link, and phlegm and fluid retention are important pathological products. For a long time, TCM treatment of chronic heart failure has primarily employed methods of tonifying qi and warming yang, promoting blood circulation, and eliminating water retention.

[0004] Over the years, continuous research in modern medicine on various aspects of CHF has gradually changed its treatment concepts, and significant progress has been made in the research of new anti-heart failure drugs in recent years. From the classic "golden triangle" to the "new triangle" based on angiotensin receptor-neprilysin inhibitors (ARNI), and now to the "new quadruple therapy," the treatment of chronic heart failure has entered a new era. The 2021 ESC Guidelines for the Diagnosis and Treatment of Acute and Chronic Heart Failure added sodium-glucose cotransporter 2 inhibitors (SGLT2i) to the "new triangle," officially changing the heart failure treatment model from the "golden triangle" to the "new quadruple therapy." Although the application of the "new quadruple therapy" can further improve the prognosis of heart failure patients and reduce the risk of sudden cardiac death, the overall efficacy is still not satisfactory, and adverse reactions such as decreased heart rate, electrolyte disturbances, and renal impairment exist.

[0005] Traditional Chinese medicine (TCM) has demonstrated significant advantages in treating chronic and intractable diseases, gaining widespread attention and increasing recognition. Based on the holistic diagnostic theory of TCM, compound herbal formulas play a crucial role in the prevention and treatment of cardiovascular diseases by promoting the functional recovery of various organs, regulating internal environmental imbalances, and reducing adverse reactions. Summary of the Invention

[0006] The purpose of this invention is to provide a traditional Chinese medicine composition for treating chronic heart failure.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] A traditional Chinese medicine composition for treating chronic heart failure is made from the following raw materials in the indicated weight ratios: Astragalus membranaceus 15–25 parts, Aconitum carmichaelii 8–10 parts, Codonopsis pilosula 8–10 parts, Cervi cornu 8–10 parts, Angelica sinensis 4–8 parts, Paeonia lactiflora 4–8 parts, Lepidium apetalum 8–12 parts, Atractylodes macrocephala (fried) 4–8 parts, Alisma plantago-aquatica 4–8 parts, Citrus aurantium 4–8 parts, Poria cocos 4–8 parts, Carthamus tinctorius 2–4 parts, and Cinnamomum cassia 4–8 parts.

[0009] To further realize the present invention, the above-mentioned traditional Chinese medicine composition is preferably made from the following raw materials in the following weight ratios: 20 parts Astragalus membranaceus, 9 parts Aconitum carmichaelii, 9 parts Codonopsis pilosula, 9 parts Cervi cornu, 6 parts Angelica sinensis, 6 parts Paeonia lactiflora, 10 parts Lepidium apetalum, 6 parts Atractylodes macrocephala (fried), 6 parts Alisma plantago-aquatica, 6 parts Citrus aurantium, 6 parts Poria cocos, 3 parts Carthamus tinctorius, and 6 parts Cinnamomum cassia.

[0010] To further realize the present invention, the above-mentioned traditional Chinese medicine composition for treating chronic heart failure is preferably prepared into decoction, pill, tea, oral liquid, tablet, capsule, granule, ointment or powder dosage form.

[0011] This invention provides an analysis of the etiology, pathogenesis, and treatment principles of chronic heart failure:

[0012] The formulation of this invention is based on the inventors' and team's many years of clinical experience and modern medical verification. They believe that "Yang deficiency" accounts for a large part of the pathogenesis of chronic heart failure, namely "chronic heart failure with Yang deficiency and blood stasis syndrome". This is because chronic heart failure is caused by various structural and other functional disorders of the cardiac system. It is often caused by ventricular blood filling and / or massive ejection, which severely impairs cardiac function and fails to fully meet the metabolic needs of the body's cells and tissues. Abnormal cardiopulmonary blood circulation is the main clinical manifestation of this disease, which falls under the category of "phlegm retention", "edema", "asthma" and "palpitation" in traditional Chinese medicine. Its location is in the heart and is closely related to the lungs, spleen and kidneys. Traditional Chinese medicine believes that chronic heart failure is a syndrome of deficiency in the root and excess in the branch. Deficiency in the root means deficiency of heart Qi and Yang, or deficiency of heart Yin. Excess in the branch means blood stasis, water retention and phlegm. Deficiency of heart Qi and Yang is the internal basis of heart failure, blood stasis is the central pathological link, and phlegm and water retention are important pathological products.

[0013] The formulation principle and compatibility rules of the traditional Chinese medicine composition for treating chronic heart failure provided by this invention are as follows:

[0014] (I) Formula Analysis

[0015] Chronic heart failure with "Yang deficiency and blood stasis syndrome" is caused by deficiency of Yang Qi, blood stasis and fluid retention. The "Suwen·Yin Yang Impression Theory" states: "Yang transforms into Qi, Yin forms into shape." Yang deficiency leads to dysfunction of Qi transformation and obstruction of the distribution of body fluids.

[0016] (II) Compatibility Rules

[0017] In this formula, Astragalus membranaceus tonifies Qi and raises Yang, promotes diuresis and reduces swelling; Aconitum carmichaelii warms the kidneys and assists Yang, transforms Qi and promotes water metabolism, together serving as the principal herbs. Codonopsis pilosula strengthens the spleen and benefits the lungs, assisting Astragalus membranaceus in tonifying vital energy; Cervi cornu warms and tonifies the liver and kidneys, nourishes essence and blood; Angelica sinensis tonifies and invigorates blood; Paeonia lactiflora clears heat and cools the blood, disperses blood stasis and relieves pain. The combination of these two herbs enhances the effects of invigorating blood, removing blood stasis, unblocking collaterals and relieving pain. Lepidium apetalum drains the lungs, relieves asthma, promotes diuresis and reduces swelling. In addition, its cold nature can also counteract the pungent and drying properties of Aconitum carmichaelii that can damage body fluids; Atractylodes macrocephala strengthens the spleen and tonifies Qi, dries dampness and promotes diuresis. The combined use of these herbs can assist the principal herbs in tonifying Qi, warming Yang, removing blood stasis and promoting diuresis, thus serving as the assistant herbs. Alisma plantago-aquatica promotes diuresis and eliminates dampness, clears heat, resolves turbidity, and lowers lipids; Citrus aurantium breaks up qi stagnation, eliminates phlegm and dissipates masses, opens the chest and promotes qi circulation, relieving chest qi stagnation and allowing qi and blood to flow smoothly; these two herbs regulate qi and guide stagnation, promote lung qi circulation, disperse stagnation and resolve phlegm, and promote edema, thus achieving the effect of harmonizing the heart and lungs; Poria cocos promotes diuresis and eliminates dampness, strengthens the spleen and calms the mind, thus enhancing the spleen-strengthening and diuretic effects of Astragalus membranaceus. Carthamus tinctorius invigorates blood circulation, unblocks menstruation, disperses blood stasis and relieves pain; Cinnamomum cassia warms and unblocks the meridians, assists yang qi transformation, calms the rebellious qi, and assists Aconitum carmichaelii in warming and tonifying kidney yang and promoting yang qi transformation; at the same time, it can guide the medicine to the heart meridian. The combined effects of these herbs are to invigorate qi, warm yang, resolve blood stasis, and promote diuresis. Attached Figure Description

[0018] Figure 1 The levels of LVEF, LVFS, and heart weight ratio in rats of each group were as follows: Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of this invention; and QFH was the high-dose group of the traditional Chinese medicine composition of this invention.

[0019] Figure 2 HE staining results of rat myocardial tissue were compared among the groups, where Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of the present invention; and QFH was the high-dose group of the traditional Chinese medicine composition of the present invention.

[0020] Figure 3 Masson staining was performed on the myocardial tissue of rats in each group, where Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of the present invention; and QFH was the high-dose group of the traditional Chinese medicine composition of the present invention.

[0021] Figure 4The levels of inflammatory factors in rats were compared among the groups, where Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of this invention; and QFH was the high-dose group of the traditional Chinese medicine composition of this invention. Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0022] Figure 5 The levels of fibrosis factors and cardiac damage indicators in rats were compared among the groups, where Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of this invention; and QFH was the high-dose group of the traditional Chinese medicine composition of this invention. Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0023] Figure 6 The expression of fibrosis-related mRNAs in rats was compared among the groups, where Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of this invention; and QFH was the high-dose group of the traditional Chinese medicine composition of this invention. Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0024] Figure 7 The expression of inflammation-related proteins in rats was compared among the groups, where Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of this invention; and QFH was the high-dose group of the traditional Chinese medicine composition of this invention. Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01.

[0025] Figure 8 The expression of fibrosis-related proteins in rats was compared among the groups, where Blank was the control group; Model was the model group; CAP was the captopril group; QFL was the low-dose group of the traditional Chinese medicine composition of this invention; and QFH was the high-dose group of the traditional Chinese medicine composition of this invention. Compared with the control group, *P<0.05, **P<0.01; compared with the model group, #P<0.05, ##P<0.01. Detailed Implementation

[0026] The present invention will be further described below with reference to specific embodiments and clinical trials.

[0027] I. Implementation Examples

[0028] Example 1:

[0029] Astragalus membranaceus 15g, Aconitum carmichaelii 8g, Codonopsis pilosula 8g, Cervi cornu 8g, Angelica sinensis 4g, Paeonia lactiflora 4g, Lepidium apetalum 8g, Atractylodes macrocephala (fried) 4g, Alisma plantago-aquatica 4g, Citrus aurantium 4g, Poria cocos 4g, Carthamus tinctorius 2g, and Cinnamomum cassia 4g.

[0030] Example 2:

[0031] Astragalus membranaceus 25g, Aconitum carmichaelii 10g, Codonopsis pilosula 10g, Cervi cornu 10g, Angelica sinensis 8g, Paeonia lactiflora 8g, Lepidium apetalum 12g, Atractylodes macrocephala (fried) 8g, Alisma plantago-aquatica 8g, Citrus aurantium 8g, Poria cocos 8g, Carthamus tinctorius 4g, and Cinnamomum cassia 8g.

[0032] Example 3:

[0033] Astragalus membranaceus 20g, Aconitum carmichaelii 9g, Codonopsis pilosula 9g, Cervi cornu 9g, Angelica sinensis 6g, Paeonia lactiflora 6g, Lepidium apetalum 10g, Atractylodes macrocephala (fried) 6g, Alisma plantago-aquatica 6g, Citrus aurantium 6g, Poria cocos 6g, Carthamus tinctorius 3g, and Cinnamomum cassia 6g.

[0034] The above-mentioned traditional Chinese medicine composition can be prepared into decoction, pills, tea, oral liquid, tablets, capsules, granules, ointment or powder dosage forms.

[0035] The method for preparing granules from the traditional Chinese medicine composition of the present invention:

[0036] Add 8 times the weight of the raw drug to water and decoct twice, 60 minutes each time. Combine the decoctions, filter, concentrate the filtrate to an appropriate amount, dry into an extract powder, add an appropriate amount of dextrin, mix well, set the drying temperature to 50-60℃, and the sieved dry granules are fed into a square cone mixer through a clean pipe for mixing for 10 minutes at a speed of 8 rpm to form granules. The granules are then dried, sized, and packaged with an inner longitudinal seal temperature of 125℃, an outer longitudinal seal temperature of 135℃, an inner transverse seal temperature of 146℃, and an outer transverse seal temperature of 146℃, with a punching speed of 68 times. The product is then obtained.

[0037] The method of taking the herbal composition of the present invention as granules is as follows: dissolve in boiling water and take 18g three times a day, with a daily dosage not exceeding 54g.

[0038] The method of taking the herbal composition of the present invention as a decoction is as follows: Prepare the above-mentioned medicines according to the formula, add 5 times the amount of water and decoct for 30 minutes each time, collect the two decoctions and concentrate to 400ml. Take 1 dose per day, divided into 2 doses, 200ml each time, and the daily dosage shall not exceed 400ml.

[0039] II. Clinical Trials

[0040] (I) Study subjects: A total of 60 patients with chronic heart failure of Yang deficiency and blood stasis type from the Affiliated Hospital of Gansu University of Traditional Chinese Medicine were selected and then randomly divided into a treatment group of 30 patients and a control group of 30 patients.

[0041] (II) Western Medicine Diagnostic Criteria:

[0042] 1. The diagnostic criteria for chronic heart failure were developed with reference to the Fmmingham study.

[0043] 2. Cardiac Function Classification Criteria: The cardiac function classification criteria were established according to the New York Heart Association (NYHA) standards revised in 1994. Patients classified as Class II, III, and IV were the primary subjects in this study.

[0044] (III) Traditional Chinese Medicine Diagnostic Criteria:

[0045] The evaluation criteria were formulated based on the "Guiding Principles for Clinical Research of New Traditional Chinese Medicines in the Treatment of Heart Failure" in the "Guiding Principles for Clinical Research of New Traditional Chinese Medicines" published by China Medical Science and Technology Press in 2002. The criteria for Yang deficiency and blood stasis syndrome in chronic heart failure are as follows:

[0046] Main symptoms: palpitations, shortness of breath, fatigue, which worsen after activity.

[0047] Secondary symptoms: spontaneous sweating, shortness of breath, cough, facial and limb edema, and oliguria.

[0048] Tongue appearance: pale and swollen tongue with ecchymosis under the tongue and white and slippery tongue coating; deep and thready pulse.

[0049] Eligibility criteria: Each participant was diagnosed by two senior TCM physicians strictly according to the above TCM syndrome diagnosis criteria. Patients meeting the following syndrome criteria must present the above primary symptoms, accompanied by at least two secondary symptoms, and the diagnosis can be made by combining tongue and pulse examination.

[0050] Inclusion criteria:

[0051] 1. All enrolled cases should meet the above diagnostic criteria for chronic heart failure;

[0052] 2. Those who are classified as Class II, III, or IV in the NYHA functional classification;

[0053] 3. Those whose TCM syndrome differentiation meets the diagnostic criteria for Yang deficiency and blood stasis syndrome;

[0054] 4. Age between 35 and 80 years old;

[0055] 5. Informed consent from the participants;

[0056] Exclusion criteria:

[0057] 1. Patients who do not meet the above inclusion criteria;

[0058] 2. Patients suffering from severe liver and kidney dysfunction, severe hematologic or endocrine system diseases, or other major systemic diseases;

[0059] 3. Patients who cannot cooperate or take medication regularly;

[0060] 4. Women who are pregnant or breastfeeding;

[0061] 5. Other risk factors: Patients with malignant arrhythmias, malignant tumors, severe infections, etc.

[0062] 6. Patients with a constitution prone to allergies or allergic reactions to medications.

[0063] (iv) Treatment methods: After enrollment, the control group received routine Western medicine treatment (sacubitril / valsartan sodium tablets + spironolactone tablets + metoprolol succinate sustained-release tablets + dapagliflozin); the treatment group received 200ml of the decoction prepared by Example 3 of the present invention orally twice daily in addition to the treatment of the control group.

[0064] Both groups underwent a 3-week treatment course, with two treatment courses in total.

[0065] (v) Observation indicators:

[0066] 1. Safety observation indicators

[0067] (1) Basic vital signs: body temperature, pulse, respiration, blood pressure;

[0068] (2) General examination items: routine blood, urine and stool tests, liver and kidney function tests, electrolyte tests and coagulation tests.

[0069] 2. Efficacy observation indicators

[0070] (1) Left ventricular ejection fraction (LVEF): Observe LVEF values ​​before and after treatment;

[0071] (2) N-terminal pro-B-type brain natriuretic peptide (NT-ProBNP): Observe the changes in NT-ProBNP before and after treatment to assess the condition of heart failure;

[0072] (3) 6-minute walk test: Observe the walking distance 6 minutes before and after treatment;

[0073] (4) Traditional Chinese Medicine symptom score;

[0074] (5) Lee's heart failure score;

[0075] (6) Cardiac function classification;

[0076] (7) Quality of life score.

[0077] (vi) Results:

[0078] (1) Comparison of general information of the two groups of patients. There was no statistically significant difference in general information between the two groups (P>0.05), as shown in Table 1.

[0079]

[0080] (2) Comparison of TCM syndrome scores between the two groups of patients before and after treatment

[0081] Before treatment, there was no statistically significant difference in TCM syndrome scores between the two groups (P>0.05). After treatment, the TCM syndrome scores of all patients decreased, but the change in the TCM group was greater than that in the Western medicine group, and the difference was statistically significant (P<0.05). The data are shown in Table 2.

[0082]

[0083] (3) The incidence of adverse reactions in the traditional Chinese medicine group was lower than that in the Western medicine group, and the difference was statistically significant (P<0.05). The data are shown in Table 3.

[0084]

[0085] (4) Before treatment, there was no significant difference in cardiac function indicators between the two groups (P>0.05); after treatment, there was a statistically significant difference between the two groups and before treatment (P<0.05), and the improvement of cardiac function indicators in the experimental group was better than that in the control group. The difference between the two groups was statistically significant (P<0.05), as shown in Table 4.

[0086]

[0087] (vii) Conclusion:

[0088] The traditional Chinese medicine composition of this invention has the effects of invigorating qi and warming yang, resolving blood stasis and promoting diuresis, and has a good clinical effect on yang deficiency and blood stasis syndrome in chronic heart failure. Compared with the control group, the TCM syndrome score of the treatment group was lower; one patient in the treatment group developed a rash and one patient developed arrhythmia, while three patients in the control group developed a rash, three patients developed drowsiness and fatigue, and two patients developed arrhythmia; the left ventricular ejection fraction increased more significantly in the treatment group than in the observation group after treatment.

[0089] III. Typical Cases

[0090] The patient, a 70-year-old female, presented to the Affiliated Hospital of Gansu University of Traditional Chinese Medicine on October 16, 2021, with a chief complaint of "recurrent chest tightness and shortness of breath for 9 years, worsening with lower extremity edema for 2 months." Nine years prior, she began experiencing chest tightness and fatigue, and had been hospitalized multiple times at local hospitals (details unknown), with symptoms fluctuating in severity. Two months prior, after a cold, she developed a cough with small amounts of white sputum, accompanied by shortness of breath on exertion and bilateral lower extremity edema, but no chest tightness or palpitations. She sought medical attention at a local hospital. A chest CT scan showed scattered inflammation in both lungs, cardiomegaly, and coronary artery calcification. After treatment with anti-infectives, bronchodilators, and other medications, her cough and sputum improved, but she still experienced shortness of breath on exertion and bilateral lower extremity edema. Past medical history: type 2 diabetes for 20 years, hypertension for 10 years, and percutaneous coronary intervention for 5 years. Presenting symptoms: cough with sputum, chest tightness and shortness of breath, wheezing upon exertion, aversion to cold and cold limbs, bilateral lower extremity edema, pale and swollen tongue with teeth marks, petechiae and ecchymoses on the edges, white and greasy tongue coating, deep, thready and slow pulse. Physical examination: blood pressure 107 / 60 mm Hg, heart rate 62 bpm. Auxiliary examinations: echocardiography showed left atrial enlargement, mild tricuspid regurgitation, mild pulmonary hypertension, and decreased left ventricular diastolic function, with an ejection fraction of 36%. N-terminal pro-brain natriuretic peptide (NT-proBNP) 438 pg / mL. Western medicine diagnosis: chronic heart failure (New York Heart Association NYHA class III), coronary heart disease, hypertension, type 2 diabetes. Traditional Chinese medicine diagnosis: heart edema, yang deficiency and blood stasis type. Treatment principle: tonify qi and warm yang, resolve blood stasis and promote diuresis. Seven doses of the herbal composition of this invention, prepared in the proportions specified in Example 2, were administered orally as a decoction.

[0091] Second visit on October 23, 2021: The patient had no cough or phlegm, felt relief from chest tightness and shortness of breath, and experienced a significant reduction in edema in both lower limbs. She still felt cold, but had no other significant discomfort. Blood pressure was 115 / 70 mm Hg, and heart rate was 65 beats / min. Her tongue was pale and swollen with a thin, white, greasy coating, and her pulse was deep and slow. She was prescribed 14 doses of the herbal composition from Example 3 of this invention, decocted in water and taken orally.

[0092] Third visit on November 13, 2021: The patient reported slight chest tightness and shortness of breath, but could meet basic daily living needs; edema in both lower extremities had subsided. Blood pressure was 110 / 70 mm Hg, heart rate 70 bpm. Tongue was pale and swollen with a thin white coating; pulse was deep and slow, but stronger than before. Repeat echocardiography revealed left atrial enlargement, mild tricuspid regurgitation, mild pulmonary hypertension, and reduced left ventricular diastolic function with an ejection fraction of 42%. The patient was prescribed the herbal composition of Example 1 of this invention, in the indicated weight proportions, and continued to take the medication.

[0093] Note: The patient is an elderly female. Due to the long duration of the illness, the yang qi of the heart and kidneys is damaged. The kidney yang is deficient and unable to receive qi and promote water metabolism. Therefore, symptoms such as chest tightness, shortness of breath, and increased气喘 with movement, as well as bilateral lower limb edema, are observed. The deficiency of yang qi in the heart and kidneys fails to warm the whole body, resulting in aversion to cold and cold limbs. The deficiency and decline of yang qi in the heart and kidneys cannot warm the spleen earth, and the spleen earth is unable to transform and transport water and dampness. The pathogenic factors of phlegm, retained fluid, and dampness invade the heart and lungs upward, leading to cough and expectoration. The tongue is pale, plump with tooth marks, there are ecchymotic spots and patches on the edges, the tongue coating is white and greasy, and the pulse is deep, thready, and slow. This is a syndrome of deficiency of heart and kidney yang, blood stasis, and retention of water. Therefore, it is necessary to benefit qi and warm yang, activate blood circulation, and promote diuresis.

[0094] In the formula, pilose antler warms the kidney and aids yang; safflower promotes blood circulation, dredges the channels, dispels stasis, and relieves pain; black processed aconite restores yang from collapse, supplements fire, and aids yang;

[0095] Astragalus membranaceus replenishes qi, raises yang, promotes diuresis, and reduces swelling; Codonopsis pilosula strengthens the spleen and benefits the lungs; Atractylodes macrocephala strengthens the spleen, replenishes qi, dries dampness, and promotes diuresis; Angelica sinensis nourishes blood, promotes blood circulation, regulates menstruation, and relieves pain; Red peony root clears heat and cools the blood, dispels stasis, and relieves pain; Lepidium apetalum purges the lung, relieves asthma, promotes diuresis, and reduces swelling; Alisma orientale promotes blood circulation to remove blood stasis, promotes diuresis, and reduces swelling; Immature bitter orange breaks qi, reduces accumulation, dissipates phlegm, and disperses stagnation; Poria cocos promotes diuresis, percolates dampness, strengthens the spleen, and tranquillizes the mind; Cinnamon twig warms the channels, aids yang in transforming qi, and calms the adverse qi.

[0096] IV. Animal experiments

[0097] 1 Materials and methods

[0098] 1.1 Animals

[0099] 60 SPF-grade male SD rats, 6 - 8 weeks old, with a body weight of (200 ± 20) g, were purchased from Beijing Spearf Bio-Technology Co., Ltd. The experimental animal license number is: SCXK (Beijing) 2019 - ********.

[0100] 1.2 Drugs

[0101] The weight ratio in Example 3 of the traditional Chinese medicine composition of the present invention was adopted.

[0102] The crude drugs were decocted twice with 8 times the weight of water for 60 minutes each time. The decoctions were combined, filtered, and the filtrate was concentrated to an appropriate amount and dried into an extract powder. An appropriate amount of dextrin was added and mixed evenly. The drying temperature was set at 50 - 60 °C. The sieved dry granules entered the conical mixer through a clean pipeline for mixing. The total mixing was carried out for 10 minutes at a rotation speed of 8 r / min to form granules, which were dried, sized, and packaged. The inner longitudinal sealing temperature was 125 °C, the outer longitudinal sealing temperature was 135 °C, the inner transverse sealing temperature was 146 °C, the outer transverse sealing temperature was 146 °C, and the punching speed was 68 times. They were divided into packages of 18 g each (委托生产 by Gansu Lanyao Pharmaceutical Co., Ltd., product batch number 20211001); Captopril tablets: (25 mg / tablet, produced by Shanxi Jinhua Huixing Pharmaceutical Co., Ltd., batch number: H19993357); DOX (100 mg / bottle, produced by Beijing Solarbio Science & Technology Co., Ltd., batch number: 25316 - 40 - 9 specification).

[0103] 1.3 Main reagents and instruments It should be noted that the "气喘" in the original text seems to be an incorrect or incomplete expression. I translated it as "increased气喘" for the sake of literal translation. You may need to check and correct it according to the actual situation. Also, the license number in item 18 is masked with asterisks as the original text "2019 - 0010" may be incomplete or incorrect in the context. You should provide the correct license number for a more accurate translation.

[0104] Interleukin-6 (IL-6), interleukin-17 (IL-17), tumor necrosis factor-α (TNF-α), transforming growth factor-α β1 (transforminggrowth factor-β, TGF- β1 Enzyme-linked immunosorbent assay kits for growth-stimulation expressed gene 2 (ST2), N-terminal pro-B-type natriuretic peptide (NT-proBNP), galectin-3 (Gal-3), and creatine kinase isoenzymes (CK-MB) (batch numbers JM-01597R2, JM-01495R2, JM-01587R2, JM-10950R2, JM-01736R2, JM-11290R2, JM-02031R2, Jiangsu Jingmei Biotechnology Co., Ltd.). Anhydrous ethanol, chloroform, isopropanol, and skim milk powder (catalog number: 508G053) were purchased from Beijing Solarbio Biotechnology Co., Ltd.; PVDF membrane (catalog number: IRVH00010) was purchased from Millipore; ECL luminescence buffer and protein electrophoresis markers (catalog numbers: 36208-A / B, 26616) were purchased from Shanghai Yisheng Biotechnology Co., Ltd.; high-performance RIPA lysis buffer, BCA protein quantification kit, and TBST buffer (catalog numbers: 20220927, 20220910, 20220917) were purchased from Beijing Solarbio Biotechnology Co., Ltd.; anti-rabbit IL-6 antibody, IL-17 antibody, TNF-α antibody, α-SMA (α-smooth muscle actin) antibody, COL-1 (Collagen I) antibody, and Tubulin antibody were all purchased from Affinity. Biosciences Ltd. (lot numbers DF6087, BF8019, AF7014, AF1032, AF7001, AF7011, Affinity Biosciences Ltd.); Anti-rabbit TGF-β1 antibody, Smad3 antibody, Smad7 antibody (lot numbers 21898-1-AP, 66516-1-Ig, 25840-1-AP, Proteintech Ltd.).

[0105] Micro-volume nucleic acid analyzer (Beijing Kai'ao Technology Development Co., Ltd., model: K5800); PCR amplification instrument (Hangzhou Langji Scientific Instruments Co., Ltd., model: A300); Real-time fluorescence PCR instrument (ROCGENE, model: 201901003); Shaker (SCILOGEX, model: SK-O180-E); Fully automated chemiluminescence imaging system (Beijing Liuyi Biotechnology Co., Ltd., model: WD-9423C); Complete electrophoresis transfer system (Bio-Rad, USA); High-speed refrigerated centrifuge (Sichuan Shuke Instrument Co., Ltd., model: TGL-1850); Microplate reader (Nanjing Detie Experimental Equipment Co., Ltd., model: HBS-1096A).

[0106] 1.4 Modeling, grouping, and drug administration

[0107] 1.4.1 CHF rat modeling method: DOX was diluted to 2 mg / mL with 0.9% sodium chloride solution. -1 After solution, 2.5 mL·kg -1 Administer intraperitoneally once a week for 6 consecutive weeks, adjusting the total DOX dose according to rat body weight to achieve a total dose of 30 mg / kg. -1 .

[0108] 1.4.2 Experimental Grouping and Intervention Methods: After one week of acclimatization, 60 rats were randomly assigned to a control group (n=12) and the remaining 48 rats were used to establish a CHF rat model as described in 1.4.1. At the end of the 6th week, the successfully modeled rats were randomly divided into a control group, a model group, a captopril group, and low- and high-dose groups of the herbal composition of this invention (n=12 each). Starting on the second day after successful model establishment, rats were administered the conventional dose, half-dose, and double-dose dose by gavage. Based on the equivalent dose conversion formula between animals and humans, the captopril group was administered 22.5 mg / (kg·d) by gavage. The low-dose herbal composition of this invention was administered at 2.84 g / (kg·d), and the high-dose herbal composition was administered at 11.34 g / (kg·d). The blank model group was administered an equal volume of sterile distilled water by gavage once daily for 6 consecutive weeks.

[0109] 1.5 Observation Indicators and Methods

[0110] 1.5.1 Echocardiography: Echocardiography was performed on the rats one day before sacrifice for tissue collection by an experienced sonographer. Left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were measured, and the average values ​​over 5 cardiac cycles were taken.

[0111] 1.5.2 Determination of the organ-to-body ratio in rats: After 6 weeks of continuous drug administration, the rat heart was removed, and residual blood was rinsed off with physiological saline. The remaining fluid was gently squeezed out, and the total weight (HW) of the heart was weighed using an electronic balance. The organ-to-body ratio of the heart was calculated. Heart-to-body ratio (mg / g) = Total heart weight (HW) / Body weight (g).

[0112] 1.5.3 Evaluation of myocardial tissue morphological changes by HE staining: Left ventricular apex tissue of each group of rats was taken, fixed with formaldehyde fixative, embedded in paraffin, and sectioned (4 μm thick). HE staining was performed and observed under a microscope.

[0113] 1.5.4 Serum IL-6, IL-17, TNF-α, TGF-β, ST2, NT-proBNP, Gal-3, and CK-MB levels were measured strictly according to the operating instructions to obtain the OD values ​​of each sample. The concentrations of the samples were calculated using standard curves and regression equations.

[0114] 1.5.5 Expression of IL-6, IL-17, TNF-α, TGF-β1, Smad3, Smad7, LncRNA-TUG1, α-SMA, and COL-1 mRNA in rat myocardial tissue. RNA was extracted according to the manufacturer's instructions, and PCR was performed according to the PrimeScript™ RT Master Mix manufacturer's instructions. Primers were synthesized by Biomed Biotechnology Co., Ltd., and the primer sequences are as follows:

[0115]

[0116] Use 2 –△△Ct The method can be used to perform relative quantification of the expression of each gene.

[0117] 1.5.6 Western Blot Detection of IL-6, IL-17, TNF-α, TGF-β1, Smad3, Smad7, α-SMA, and COL-1 Protein Expression in Myocardial Tissue Apical and left ventricular tissues were lysed and extracted. Protein concentrations were quantified using a BCA kit, followed by SDS-PAGE electrophoresis. Proteins were then transferred to PVDF membranes. To prevent nonspecific binding, fetal bovine serum albumin was used for blocking, and secondary antibody was added during incubation. Finally, enhancement chemiluminescence (ECL) was used for imaging, and band grayscale analysis was performed using ImageJ 1.x software. The ratio of the bands to the internal control β-actin was used to determine the relative protein expression level.

[0118] 1.6 Statistical Methods

[0119] Statistical analysis was performed using SPSS 26.0 software. The quantitative data, after testing, met the requirements of normal distribution and homogeneity of variance, and were expressed as mean ± standard deviation. The data were processed using one-way ANOVA and multiple comparisons were performed using the Bonferroni test. The results showed that... P <0.05 indicates statistical significance. P A value <0.01 indicates a significant difference. In cases of unequal variances, the differences between groups are not statistically significant.

[0120] 2 Results

[0121] A total of 14 rats died during the gavage administration process, including 4 in the control group and 2 in each of the other groups. 8 rats survived in each group.

[0122] 2.1 Effects of the herbal composition of the present invention on cardiac function and organ-to-body ratio in rats with heart failure

[0123] like Figure 1 As shown, compared with the control group, the LVEF, LVFS levels and heart weight ratio of rats in the model group were decreased (P<0.05); compared with the model group, the LVEF, LVFS levels and heart weight ratio of rats in the captopril group, the low-dose group and the high-dose group of the herbal composition of the present invention were increased (P<0.05), indicating that the herbal composition of the present invention can alleviate myocardial fibrosis in rats with heart failure.

[0124] 2.3 Results of HE staining and Masson staining of rat myocardial tissue in each group

[0125] like Figure 2 As shown, compared with the control group, the model group showed a small number of hypertrophic cardiomyocytes with deeply stained nuclei, some cardiomyocytes with degeneration and necrosis, muscle fiber breakage, and interstitial inflammatory cell infiltration; the captopril group showed some hypertrophic cardiomyocytes with deeply stained nuclei, and no obvious interstitial inflammatory cell infiltration; the low-dose group of the herbal composition of the present invention showed a small number of hypertrophic cardiomyocytes with deeply stained nuclei, some cardiomyocytes with pyknosis, loose muscle fiber arrangement, and interstitial inflammatory cell infiltration; the high-dose group of the herbal composition of the present invention showed cardiomyocytes with basically normal cell morphology and no obvious interstitial inflammatory cell infiltration.

[0126] like Figure 3 As shown, compared with the control group, the cross-sectional area and ventricular cavity area of ​​the myocardial tissue stained with Masson's staining in the model group rats were significantly reduced, and myocardial fibrosis of varying degrees was observed (P<0.01). Compared with the model group, the cross-sectional area and ventricular cavity area of ​​the myocardial tissue stained with Masson's staining in the captopril group, and the low- and high-dose groups of the herbal composition of this invention were increased, and the area of ​​myocardial fibrosis was reduced (P<0.05).

[0127] 2.4 Serum expression in each group of rats

[0128] The ELISA kit was used to detect factors in rat serum. The results showed that, compared with the control group, the model group rats had higher levels of inflammatory factors IL-6, IL-17, Gal-3, and TNF-α, and the fibrosis marker TGF-β. β 1. Levels of ST2, CK-MB, and the heart failure marker NT-proBNP were all elevated. P< 0.05); compared with the model group, the captopril group, and the low- and high-dose groups of the traditional Chinese medicine composition of this invention showed lower levels of IL-6, IL-17, Gal-3, TNF-α, and TGF-β in rats. β 1. ST2, CK-MB, and NT-proBNP levels all decreased ( P< 0.05), indicating that the herbal composition of the present invention can reduce the serum inflammation level in the model group rats, see Figure 4 , 5 .

[0129] 2.5 mRNA expression in myocardial tissue of rats in each group

[0130] Compared with the control group, the model group rats had higher levels of IL-6, IL-17, TNF-α, LncRNA-TUG1, and TGF-β in their myocardial tissue. β 1. The expression levels of Smad3, α-SMA, and COL-1 mRNA were all increased ( P< 0.01), Smad7 mRNA expression level decreased ( P< 0.01); Compared with the model group, the captopril group, and the low- and high-dose groups of the traditional Chinese medicine composition of this invention showed lower levels of IL-6, IL-17, TNF-α, LncRNA-TUG1, and TGF-β in the myocardial tissue of rats. β 1. The expression levels of Smad3, α-SMA, and COL-1 mRNA were all decreased. P< 0.05), Smad7 mRNA expression level increased ( P< 0.05), indicating that the herbal composition of the present invention alleviates myocardial fibrosis in rats by inhibiting the expression of inflammation-related protein mRNA, reducing extracellular matrix deposition, see [reference needed]. Figure 6 .

[0131] 2.6 Verification of myocardial proteins and expression of fibrosis-related tissues in each group of rats

[0132] Compared with the control group, the model group rats had higher levels of inflammatory proteins IL-6, IL-17, and TNF-α, as well as the fibrotic factor TGF-β in their myocardial tissue. β 1. The expression levels of Smad3, Smad7, α-SMA, and COL-1 proteins were all increased. P<0.01), Smad7 protein expression level decreased ( P< 0.01); Compared with the model group, the captopril group, and the low- and high-dose groups of the traditional Chinese medicine composition of this invention showed lower levels of IL-6, IL-17, TNF-α, and TGF-β in the myocardial tissue of rats. β 1. The expression levels of Smad3, α-SMA, and COL-1 proteins were all decreased. P< 0.05), Smad7 protein expression level increased ( P< 0.05), indicating that the herbal composition of the present invention alleviates myocardial fibrosis in rats by inhibiting the expression of inflammation-related proteins and reducing extracellular matrix deposition. Figure 7 , 8 .

Claims

1. A traditional Chinese medicine composition for treating chronic heart failure, characterized in that... It is made from the following ingredients in the indicated weight proportions: Astragalus membranaceus 15–25 parts, Aconitum carmichaelii 8–10 parts, Codonopsis pilosula 8–10 parts, Cervi cornu 8–10 parts, Angelica sinensis 4–8 parts, Paeonia lactiflora 4–8 parts, Lepidium apetalum 8–12 parts, Atractylodes macrocephala (fried) 4–8 parts, Alisma plantago-aquatica 4–8 parts, Citrus aurantium 4–8 parts, Poria cocos 4–8 parts, Carthamus tinctorius 2–4 parts, and Cinnamomum cassia 4–8 parts.

2. The traditional Chinese medicine composition for treating chronic heart failure as described in claim 1, characterized in that... It is made from the following ingredients in the indicated weight proportions: Astragalus membranaceus 20 parts, Aconitum carmichaelii 9 parts, Codonopsis pilosula 9 parts, Cervi cornu 9 parts, Angelica sinensis 6 parts, Paeonia lactiflora 6 parts, Lepidium apetalum 10 parts, Atractylodes macrocephala (fried) 6 parts, Alisma plantago-aquatica 6 parts, Citrus aurantium 6 parts, Poria cocos 6 parts, Carthamus tinctorius 3 parts, and Cinnamomum cassia 6 parts.

3. The traditional Chinese medicine composition for treating chronic heart failure as described in claim 1 or 2, characterized in that: It can be made into decoctions, pills, teas, oral liquids, tablets, capsules, granules, ointments, or powders.

Images