Use of Xinshubao tablets in preparing medicines for preventing and / or treating heart failure

Abstract

The invention discloses the use of Xinshubao tablets in preparing drugs for preventing and / or treating heart failure. Animal experiments prove that Xinshubao tablets can significantly prevent and treat heart failure by improving cardiac function and reducing cardiac fibrosis. Xinshubao tablets are applied to heart failure, especially to the prevention and treatment of heart failure with mildly reduced ejection fraction or heart failure with reduced ejection fraction, and have broad application prospects.

Description

Technical Field

[0001] The present invention specifically relates to the use of Xinshubao tablets in preparing medicines for preventing and / or treating heart failure. Background Art

[0002] Heart failure (HF) is growing significantly worldwide with an aging population, with a clinical incidence of 2% and an annual mortality rate of nearly 5%. Treatment as early as possible is crucial to improving heart failure and improving the quality of life, but there is currently a lack of effective early warning markers and effective drugs for early treatment. According to the 2022 ACC\AHA\HSFA clinical diagnosis and treatment guidelines, the existing "new quadruple" treatment model drugs are all blockers that mainly target target groups represented by sodium-glucose co-transporter 2 inhibitors (SGLT2i), and the combination of drugs has certain interactions and side effects, which increases the adverse reactions of some patients and threatens the physical and mental health of patients. Therefore, preventing and treating the early occurrence and development of heart failure is the important challenge currently faced.

[0003] Xinshubao Tablets are a drug prepared by Zhangzhou Pien Tze Huang Pharmaceutical Co., Ltd. for the treatment of coronary heart disease, chest tightness caused by qi deficiency and blood stasis, angina pectoris, hypertension, hyperlipidemia and arteriosclerosis, etc. It is composed of salvia miltiorrhiza, white peony root, curcuma, Acanthopanax senticosus and hawthorn. There is no record of the treatment of early heart failure in its instructions approved by the drug regulatory authorities, and there is no research on the application of Xinshubao Tablets in the treatment of heart failure, especially mildly reduced ejection fraction or reduced ejection fraction heart failure. Summary of the invention

[0004] To solve the above problems, the present invention provides a composition for use in preparing a drug for preventing and / or treating heart failure, wherein the composition is prepared from the following raw materials in parts by weight:

[0005] 100-300 parts of Salvia miltiorrhiza, 1000-3000 parts of White Peony Root, 150-450 parts of Curcuma Radix, 500-1500 parts of Acanthopanax Senticosus, and 1000-3000 parts of Crataegus.

[0006] Furthermore, the drug is a drug for preventing and / or treating heart failure with mild reduced ejection fraction or heart failure with reduced ejection fraction.

[0007] Furthermore, the drug is a drug for improving heart function.

[0008] Furthermore, the drug is a drug that increases cardiac ejection fraction and fractional shortening.

[0009] Furthermore, the drug is a drug that regulates the expression of ANP and BNP factors in cardiac serum.

[0010] Furthermore, the drug is a drug for alleviating cardiac fibrosis.

[0011] Furthermore, the drug is a drug that reduces collagen fiber deposition in the heart.

[0012] Furthermore, the drug is a drug that reduces cardiac fibrosis-related proteins ARRB1 and THBD. Furthermore, the drug is a drug that increases cardiac fibrosis-related protein FGF1.

[0013] Furthermore, the composition is prepared from the following raw materials in parts by weight:

[0014] 200 parts of Salvia miltiorrhiza, 2000 parts of White Peony Root, 300 parts of Curcuma Radix, 1000 parts of Acanthopanax Senticosus, and 2000 parts of Crataegus.

[0015] Furthermore, the medicine is a preparation prepared with the composition as an active ingredient and pharmaceutically acceptable excipients; the preparation is an oral preparation.

[0016] Furthermore, the oral preparation is preferably Xinshubao tablets.

[0017] The "heart failure with mildly reduced ejection fraction" mentioned in the present invention refers to heart failure with a left ventricular ejection fraction of 41%-49%.

[0018] The "heart failure with reduced ejection fraction" mentioned in the present invention refers to heart failure with a left ventricular ejection fraction of ≤40%.

[0019] The invention discloses a method for preparing a drug for preventing and / or treating heart failure. Animal experiments show that the Xinshubao tablets can significantly prevent and treat heart failure by improving cardiac function and reducing cardiac fibrosis. The Xinshubao tablets are applied to heart failure, especially to the prevention and treatment of heart failure with mildly reduced ejection fraction or heart failure with reduced ejection fraction, and have broad application prospects.

[0020] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0021] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The results of dynamic ejection fraction evaluation of heart failure mice by cardiac ultrasound;

[0023] Figure 2 The results of cardiac ultrasound evaluation of the effect of Xinshubao tablets on the ejection fraction of heart function in mice with heart failure;

[0024] Figure 3 The results of cardiac ultrasound evaluation of the effect of Xinshubao tablets on the short-axis shortening rate of heart function in mice with heart failure;

[0025] Figure 4 HE staining was used to detect the effect of Xinshubao tablets on the cardiac tissue morphology of mice with heart failure;

[0026] Figure 5 ELISA test results of the effect of Xinshubao tablets on the expression of ANP in heart failure mice;

[0027] Figure 6 ELISA test results of the effect of Xinshubao tablets on BNP expression in heart failure mice;

[0028] Figure 7 Masson staining was used to detect the effect of Xinshubao tablets on the degree of cardiac tissue fibrosis in mice with heart failure;

[0029] Figure 8 Western blotting to detect the effect of Xinshubao tablets on the expression of THBD in the heart of mice with heart failure;

[0030] Fig. 9 Western blotting to detect the effect of Xinshubao tablets on the expression of FGF1 in the heart of mice with heart failure;

[0031] Fig.10 Western blotting results show the effect of Xinshubao tablets on the expression of ARRB1 in the heart of mice with heart failure. DETAILED DESCRIPTION

[0032] The raw materials, equipment and reagents used in the specific embodiments of the present invention are all known products, which are obtained by purchasing commercially available products. Among them, Xinshubao tablets are provided by Zhangzhou Pien Tze Huang Pharmaceutical Co., Ltd., which are prepared from Salvia miltiorrhiza, White Peony Root, Curcuma Radix, Acanthopanax Senticosus and Crataegus Pinellia with a mass ratio of 200:2000:300:1000:2000 as raw materials.

[0033] Example 1 Study on the application of Xinshubao tablets in treating heart failure

[0034] 1. Methods

[0035] 1. Establishment and evaluation of mouse models of heart failure with mild or reduced ejection fraction

[0036] (1) Animal grouping: 110 adult male C57BL / 6J mice were randomly divided into 4 groups, 27 mice in each group: Group 1 was Sham group (sham operation group); Group 2 was Model group (myocardial infarction model group); Group 3 was XSB-L group (Model + Xinshubao tablets low-dose group); Group 4 was XSB-H group (Model + Xinshubao tablets high-dose group). 24 hours after LAD ligation, the male C57BL / 6N mice were continuously gavaged with pure water in the Sham group and the Model group, the Model + Xinshubao tablets low-dose group was administered with a concentration of 0.026 g / ml and a dosage of 0.26 g / kg, and the Model + Xinshubao tablets high-dose group was administered with a concentration of 0.052 g / ml and a dosage of 0.52 g / kg. After that, gavage was continued for 56 days (8 weeks), once a day.

[0037] (2) Modeling of heart failure with mild or reduced ejection fraction: Male C57BL / 6J mice were anesthetized by intraperitoneal injection of 1% sodium pentobarbital and fixed on the operating table. The mice were intubated and supported by a ventilator with a frequency of 120 times / min, an inspiratory-expiratory ratio of 1:1, and a tidal volume of 1.7. The chest cavity was opened at the axilla of the left forelimb with ophthalmic scissors and at the third and fourth intercostal spaces with microscissors to expose the left anterior descending coronary artery (LAD) of the heart. The needle was inserted horizontally 2 to 3 mm from the lower edge of the left atrial appendage, ligated, and the chest cavity was closed.

[0038] 2. Evaluation of the efficacy of Xinshubao tablets on mice with heart failure

[0039] (1) Cardiac ultrasound examination: 8 weeks after surgery, the mice were placed in an anesthesia box with 5% isoflurane for anesthesia, fixed on the operating table in the supine position, and maintained with 1% isoflurane for anesthesia. The parasternal short-axis section was marked at the level of the left ventricular papillary muscle, and the left ventricular ejection fraction (LVEF), left ventricular short-axis shortening rate (LVFS), etc. were recorded using M-mode ultrasound mode. The parasternal long-axis section was marked at the level of the left ventricular outflow tract, and the ventricular wall motion image was recorded using B-mode echocardiography.

[0040] (2) Hematoxylin-eosin staining to detect morphological changes in mouse heart tissue: The experiment was terminated 8 weeks after surgery. The mouse eyeballs were removed to collect blood, and the serum was collected by centrifugation. The heart tissue samples were collected and stored at -80 degrees and fixed with 4% paraformaldehyde. After paraffin embedding, the slice thickness was about 6 μm. The paraffin sections were dewaxed to water. The sections were stained with hematoxylin stain for 3-5 minutes, washed with tap water, differentiated with differentiation solution, washed with tap water, blued with blue regeneration solution, and rinsed with running water. The sections were dehydrated in 85% and 95% gradient alcohol for 5 minutes each, and stained in eosin stain for 5 minutes. Dehydrated and sealed with neutral gum. Microscopic examination, image acquisition and analysis. Under the microscope, the cell nucleus was stained blue and the cytoplasm was stained red.

[0041] (3) ELISA detection of ANP and BNP content in mouse serum: Take blood samples from mouse ocular veins, store at room temperature for 2 hours, centrifuge at 2000rpm for 10 minutes, and take the supernatant. Set standard wells, blank wells and sample wells respectively, add 50ul of diluted standard solution to the standard wells, add 50ul of sample diluent to the blank wells, add 50ul of the sample to be tested to the remaining wells, and incubate in a 37° oven for 60 minutes. After discarding the liquid in the wells, add 50ul of biotin antibody working solution, cover the ELISA plate and incubate in a 37° oven for 45 minutes, discard the liquid in the wells, add 350ul of washing solution to each well, soak for 2 minutes, discard the liquid in the wells and pat dry on thick absorbent paper. After repeating the washing steps 3 times, add 100ul of enzyme conjugate working solution to each well, cover the plate and incubate in a 37° oven for 30 minutes. After discarding the enzyme conjugate working solution in the well, add 350ul of washing solution and soak for 2min, discard the washing solution in the well and pat dry, repeat the washing step 5 times, add 90ul of substrate solution to each well, cover the film and incubate for 15min (37°, avoid light). After adding 50ul of stop solution, immediately use an ELISA reader to measure the absorbance value of each well at 450nm. The levels of ANP and BNP in mouse serum were detected using ELISA kits from Wuhan Huamei and Elabscience according to the instructions.

[0042] 3. Evaluation of the effect of Xinshubao tablets on improving cardiac fibrosis in mice with heart failure

[0043] (1) Masson staining: The embedded tissue sections were routinely dewaxed and stained with prepared hematoxylin for 3 min, washed with water and differentiated with differentiation solution for 3 s, rinsed with tap water and then blued with bluing solution, rinsed with tap water and then stained with ponceau acid fuchsin solution for 8 min, rinsed with tap water and then soaked in phosphomolybdic acid for 3 min and then directly stained with aniline blue for 3 min; soaked in 1% glacial acetic acid for 1 min, dehydrated and sealed, and the morphological changes were observed and photographed using a panoramic scanner of a digital sectioning system.

[0044] (2) Western blotting to detect the expression of cardiac fibrosis proteins in mice: Extract proteins from the left ventricle of mice, add them to 1×RIPA buffer containing protease inhibitors, grind them with a homogenizer, and centrifuge to obtain the supernatant. After the concentration is determined, the extracted proteins are electrophoresed on a 10% SDS-polyacrylamide gel and transferred to a polyvinylidene fluoride (PVDF) membrane through an electrotransfer process. Block the membrane with 5% skim milk powder (dissolved in 1X TBST) for 1 hour. After the PVDF membrane is blocked, it is incubated with specific antibodies (ARRB1, FGF1 and THBD) overnight. After washing with TBST, it is incubated with goat anti-rabbit or goat anti-mouse IgG-horseradish peroxidase (HRP)-coupled secondary antibodies at room temperature for another 1 hour. After washing with TBST, it is detected with enhanced chemiluminescence (ECL) detection reagent.

[0045] 2. Results

[0046] 1. Evaluation of the efficacy of Xinshubao tablets on mice with mild or reduced ejection fraction heart failure

[0047] According to the 2022 ACC\AHA\HSFA clinical diagnosis and treatment guidelines, heart failure with reduced LVEF (HFrEF) is defined as LVEF ≤ 40%; heart failure with mildly reduced LVEF (HFmrEF) is defined as LVEF = 41%-49%. During the 1-4 weeks after modeling, LVEF was found to be 41%-49%, and heart failure characteristics of LVEF ≤ 40% were observed after 4 weeks. The dynamic process results of ejection fraction by echocardiography are shown in Figure 1 .

[0048] Cardiac ultrasound examination of 8-week-old heart failure mice showed that compared with the model group, Xinshubao tablets could increase the ejection fraction and short-axis shortening rate of the mice's hearts. Figure 2 and 3 As shown in the figure, the ejection fraction of the XSB-L group was (47.0837±4.20133)%, and the short axis shortening rate was (23.1690±2.41476)% (N=5), and the ejection fraction of the XSB-H group was (44.7832±1.83272)%, and the short axis shortening rate was (22.0316±0.97402)% (N=6). Hematoxylin-eosin staining was used to detect the morphological changes of mouse heart tissue, as shown in the figure. Figure 4 As shown in the figure, compared with the model group, Model + Xinshubao tablets can reduce inflammatory cell infiltration in cardiac tissue, cell nucleus condensation and other lesions, indicating that Xinshubao tablets can inhibit cardiac tissue cell necrosis. ELISA was used to detect the levels of ANP and BNP in mouse serum. The results showed that compared with the model group, the expression of ANP and BNP in the serum of mice in the high-dose Model + Xinshubao group was significantly reduced. Figure 5 and Figure 6 The above experimental results show that the effect of Xinshubao at low doses is better than that at high doses, indicating that Xinshubao can achieve excellent effects within this dose range (human equivalent dose).

[0049] In summary, Xinshubao tablets can increase cardiac ejection fraction and short-axis shortening rate, inhibit cardiac tissue cell necrosis, and regulate the expression of ANP and BNP factors in mouse heart serum, thereby improving cardiac function.

[0050] 2. Evaluation of the effect of Xinshubao tablets on improving cardiac fibrosis in mice with heart failure

[0051] Masson staining of mouse hearts showed that compared with the model group, the deposition of collagen fibers in the hearts of the high- and low-dose groups of Model+Xinshubao tablets was significantly reduced. Figure 7The expression of cardiac fibrosis proteins in mice was detected by Western blotting. Figure 8 , 9 As shown in Figures 10, compared with the model group, the cardiac fibrosis-related proteins ARRB1 and THBD in the Model+Xinshubao tablets high and low dose groups were significantly reduced, while FGF1 was significantly increased. These results indicate that Xinshubao tablets can alleviate cardiac fibrosis.

[0052] In summary, Xinshubao tablets can regulate the expression of proteins ARRB1, FGF1 and THBD involved in regulating fibrosis during cardiac remodeling, and can reduce the deposition of collagen fibers, thus having an anti-cardiac fibrosis effect.

Claims

1. Use of a composition in the preparation of a medicament for preventing and / or treating heart failure, characterized in that: The composition is prepared from the following raw materials in parts by weight: 200 parts of Salvia miltiorrhiza, 2000 parts of White Peony Root, 300 parts of Curcuma Radix, 1000 parts of Acanthopanax Senticosus, and 2000 parts of Crataegus.

2. The use according to claim 1, characterized in that: The drug is a drug for preventing and / or treating heart failure with mildly reduced ejection fraction or heart failure with reduced ejection fraction.

3. The use according to claim 1 or 2, characterized in that: The drug is a drug for improving heart function.

4. The use according to claim 3, characterized in that: The drug is a drug that increases cardiac ejection fraction and fractional shortening.

5. The use according to claim 3, characterized in that: The drug is a drug for inhibiting cardiac tissue cell necrosis and / or regulating the expression of ANP and BNP factors in cardiac serum.

6. The use according to claim 1 or 2, characterized in that: The drug is a drug for alleviating cardiac fibrosis.

7. The use according to claim 6, characterized in that: The drug is a drug that reduces collagen fiber deposition in the heart.

8. The use according to claim 6, characterized in that: The drug is a drug that reduces cardiac fibrosis-related proteins ARRB1 and THBD, and / or increases cardiac fibrosis-related protein FGF1.

9. The use according to claim 1 or 2, characterized in that: The medicine is a preparation prepared by taking the composition as the active ingredient and adding pharmaceutically acceptable excipients.

10. The use according to claim 9, characterized in that: The preparation is an oral preparation.

11. The use according to claim 10, characterized in that: The oral preparation is Xinshubao tablets.

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