Application of salvianolic acid C in preparation of medicine for preventing and treating heart failure

Abstract

The invention discloses application of salvianolic acid C in preparation of a medicine for preventing and treating heart failure. A plurality of in-vitro model results show that the salvianolic acid C can improve heart failure related indexes, increase the survival rate of myocardial cells and improve myocardial cell hypertrophy. In an in-vivo experiment, cardiac ultrasound shows that the salvianolic acid C plays a role in protecting functional impairment caused by heart failure and can improve the heart function of a mouse with heart failure to a certain extent, and HE and Masson staining results show that the salvianolic acid C can also protect the heart tissue structure and improve the pathological change of the heart tissue caused by heart failure, so that the salvianolic acid C can be used for treating heart failure. Serum biochemical results show that the salvianolic acid C has a down-regulation effect on the rise of lactic dehydrogenase (LDH), creatine kinase isoenzyme (CK-MB), cardiac troponin I (cTNI) and brain natriuretic peptide (BNP) caused by heart failure. The salvianolic acid C can be used as a medicine for preventing and treating heart failure, and has higher clinical application value and development prospect.

Description

Technical Field

[0001] The present invention belongs to the field of traditional Chinese medicine, relates to the field of preparation of anti-heart failure drugs, and particularly relates to the application of salvianolic acid C in the preparation of drugs for treating heart failure. Background Art

[0002] Heart failure (hereinafter referred to as HF) refers to symptoms and signs caused by abnormal heart structure or function due to various factors, resulting in ventricular systolic or filling disorders, and is almost the "final battlefield" for the occurrence of all cardiovascular diseases. As the terminal stage of heart diseases, the incidence of heart failure shows an increasing trend year by year with the acceleration of the aging process of the population and the increase of risk factors for cardiovascular diseases. The quality of life of HF patients has severely declined. They not only face physical symptoms such as dyspnea, fatigue, and edema, but also bear huge psychological pressure. Moreover, the readmission rate and mortality rate of HF patients remain high, bringing a heavy economic burden to the patients' families and society.

[0003] Currently, the first-line drugs for treating heart failure include angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers (ACE / ARBs), β-blockers, inotropic drugs, diuretics, and aldosterone receptor antagonists, etc. Sodium-glucose cotransporter 2 (SGLT2) inhibitors and Vericiguat (a soluble guanylate cyclase activator) have also been proven to improve the prognosis of patients. However, these drugs can only delay the progression and have a wide range of side effects, including inducing arrhythmia, causing electrolyte disorders, and aggravating renal function damage, etc. Heart failure is a multi-factor complex disease. Although modern medicine has made certain progress in the prevention and treatment of heart failure, the existing prevention and treatment strategies still cannot meet the clinical needs. Therefore, optimizing the drug treatment strategy is the key measure to extend the survival period of patients.

[0004] Salvia miltiorrhiza is the dried root and rhizome of the plant Salvia miltiorrhiza Bunge in the family Labiatae. Modern pharmacological studies have shown that Salvia miltiorrhiza has good cardio-cerebrovascular protection, and salvianolic acid C is a water-soluble phenolic acid compound extracted from Salvia miltiorrhiza, with a relatively high content and prominent activity. Modern research has shown that salvianolic acid C has various pharmacological activities such as anti-inflammatory, antioxidant, anti-fibrotic, and neuroprotective effects. However, there is no research on whether salvianolic acid C has a therapeutic effect on heart failure. Summary of the Invention

[0005] To solve the above technical problems, the present invention provides a new application of salvianolic acid C, specifically providing the application of salvianolic acid C in the preparation of drugs for treating heart failure. The molecular formula of the salvianolic acid C is: C 26 H 20 O 10 , and the structural formula is shown as follows:

[0006] Preferably, when applied, the dosage of the drug is 20-80 μM.

[0007] The research of the present invention shows that salvianolic acid C can dose-dependently reduce myocardial injury caused by doxorubicin (DOX) and cobalt chloride (Cocl2)-induced heart failure and improve isoproterenol (ISO)-induced myocardial hypertrophy within the range of 20-80 μM.

[0008] Furthermore, the present invention takes salvianolic acid C as the research object, and uses a DOX-induced heart failure model of C57BL / 6J mice to investigate the effect of salvianolic acid C on mouse heart failure. The results show that compared with the heart failure model group, salvianolic acid C at 10 mg / kg, 20 mg / kg, and 40 mg / kg has a therapeutic effect on the C57BL / 6J mouse heart failure model, can significantly improve the left ventricular function of mice (including ejection fraction EF and fractional shortening FS), improve the pathological changes of heart tissue, and reduce the levels of LDH, CK-MB, BNP, and cTNI caused by heart failure.

[0009] Preferably, the dosage form of the drug is tablet, capsule, oral liquid, pill, granule, paste, powder, mixture or syrup. The dosage form of the drug of the present invention can be diversified, and any oral preparation dosage form that can effectively deliver the active ingredient to the mammalian body is acceptable.

[0010] The drug also includes pharmaceutically acceptable excipients; the pharmaceutically acceptable excipients include any one or a combination of at least two of carrier, diluent, excipient, filler, binder, wetting agent, disintegrant, emulsifier, cosolvent, solubilizer, osmotic pressure regulator, surfactant, coating material, colorant, pH regulator, antioxidant, bacteriostatic agent or buffer.

[0011] Furthermore, the drug also includes pharmaceutically acceptable excipients. In the present invention, the "pharmaceutically acceptable" components are substances that are applicable to humans and other mammals without excessive adverse reactions (such as toxicity, irritation and allergic reactions), that is, substances with a reasonable benefit / risk ratio.

[0012] Preferably, the excipients include pharmaceutically acceptable carriers and excipients, and the carriers can be selected from starch, microcrystalline cellulose, dextrin, sterilized distilled water, normal saline, powdered sugar, etc.

[0013] Even further, the drug is an oral dosage form.

[0014] Preferably, when the application object is a mouse, the effective dose of salvianolic acid C is 10-40 mg / kg. When the application object is a human, the human dosage can be correspondingly converted according to the relevant dosing standards in the pharmaceutical field. In the case of a relatively large single dose, it can be administered in multiple doses over multiple days.

[0015] Advantages of the present invention:

[0016] (1) Research of the present invention shows that salvianolic acid C can improve the viability of cardiomyocytes in a heart failure model induced by DOX and CoCl2 in vitro and improve ISO-induced myocardial hypertrophy.

[0017] (2) Research of the present invention shows the beneficial therapeutic effect of salvianolic acid C on heart failure, which can increase the left ventricular ejection fraction and fractional shortening of heart failure mice and improve pathological phenomena such as loose arrangement of myocardial tissue, infiltration of inflammatory cells, and deposition of collagen fibers caused by heart failure.

[0018] (3) Salvianolic acid C can significantly reduce the levels of LDH, CK-MB, BNP, and cTNI caused by heart failure. Description of the Drawings

[0019] Figure 1 It shows the effect of salvianolic acid C on the survival rate of H9C2 cells in a heart failure model induced by DOX. Among them, A is the detection of cell survival rate by CCK-8; B is the representative diagram of cell state under the microscope, scale bar = 100 μm. : P < 0.001, compared with the Control group; *: P < 0.05, **: P < 0.01, ***: P < 0.001, compared with the model group.

[0020] Figure 2 It shows the effect of salvianolic acid C on the survival rate of H9C2 cells in a heart failure model induced by ischemia and hypoxia caused by CoCl2. : P < 0.001, compared with the Control group; **: P < 0.01, ***: P < 0.001, compared with the model group.

[0021] Figure 3 It shows the effect of salvianolic acid C on the cross-sectional area of H9C2 cells in a pressure overload-induced heart failure model induced by ISO. Among them, A is the representative diagram of phalloidin staining of each group, scale bar = 100 μm; B is the statistical result of the cross-sectional area of each group of cells. ##: P < 0.01, compared with the Control group; **: P < 0.01, compared with the model group.

[0022] Figure 4 It shows the results of mouse heart ultrasound examination. Among them, A is the representative diagram of echocardiogram of each group of mice; B is the statistical chart of left ventricular ejection fraction; C is the statistical chart of fractional shortening. ##: P < 0.01, : P < 0.001, compared with the Control group; *: P < 0.05, **: P < 0.01, ***: P < 0.001, compared with the model group.

[0023] Figure 5It is the HE staining result of mouse heart tissue. Scale bar = 100 μm.

[0024] Figure 6 It is the Masson staining result of mouse heart tissue. Scale bar = 100 μm.

[0025] Figure 7 It is the detection of heart failure-related indicators in mouse serum. Among them, A is the statistical chart of LDH content; B is the statistical chart of CK-MB content; C is the statistical chart of cTNI content; D is the statistical chart of BNP content. #: P < 0.05, : P < 0.001, compared with the Control group; *: P < 0.05, **: P < 0.01, ***: P < 0.001, compared with the model group. Detailed implementation manners

[0026] Next, the technical solutions of the present invention will be clearly and completely described in conjunction with the drawings and embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative efforts belong to the protection scope of the present invention.

[0027] Example 1: DOX-induced in vitro heart failure model

[0028] Taking rat H9C2 cells as the research object, 5 groups were set up, namely the control group, the DOX group, the low-dose salvianolic acid C treatment group, the medium-dose salvianolic acid C treatment group, and the high-dose salvianolic acid C treatment group. The dose of DOX was 1 μM, and the doses of salvianolic acid C were 20, 40, and 80 μM. The control group was the solvent dimethyl sulfoxide (DMSO). After co-administering for 24 h, CCK-8 staining was used to detect the survival rate of H9C2 cells, and the cell state was observed under a microscope. The calculation formula was: the survival rate of cells in the administration group = (absorbance value of the administration group - absorbance value of the blank group) / (absorbance value of the control group - absorbance value of the blank group) × 100%.

[0029] The experimental results are specifically described as follows: As Figure 1 shown, the survival rate of cells in the DOX group was 63.67% ± 6.48% (p < 0.0001), the survival rate of cells in the 20 μM salvianolic acid C treatment group was 75.43% ± 5.27% (p = 0.0126), the survival rate of cells in the 40 μM salvianolic acid C treatment group was 79.72% ± 1.37% (p = 0.0016), and the survival rate of cells in the 80 μM salvianolic acid C treatment group was 90.57% ± 1.56% (p < 0.0001). It was also observed under a microscope that the cell state had been significantly improved after administration. It shows that salvianolic acid C can significantly increase the survival rate of cardiomyocytes under the action of DOX. The results are shown in Figure 1 .

[0030] Example 2: In vitro heart failure model induced by CoCl2-induced ischemia and hypoxia

[0031] Using rat H9C2 cells as the research object, 5 groups were set up, namely the control group, the CoCl2 group, the low-dose salvianolic acid C treatment group, the medium-dose salvianolic acid C treatment group, and the high-dose salvianolic acid C treatment group. The dose of CoCl2 was 500 μM, the dose of salvianolic acid C was 20, 40, and 80 μM, and the control group was the solvent dimethyl sulfoxide (DMSO). After pre-administering salvianolic acid C for 24 h, CoCl2 was administered for 24 h, and finally, the CCK-8 staining was used to detect the survival rate of H9C2 cells. The calculation formula was: cell survival rate of the administration group = (absorbance value of the administration group - absorbance value of the blank group) / (absorbance value of the control group - absorbance value of the blank group) × 100%.

[0032] The experimental results are specifically described as follows: As Figure 2 shown, the cell survival rate of the CoCl2 group was 79.92% ± 2.55% (p = 0.0001), the cell survival rate of the 20 μM salvianolic acid C treatment group was 93.75% ± 3.17% (p = 0.0020), the cell survival rate of the 40 μM salvianolic acid C treatment group was 105.3% ± 5.67% (p < 0.0001), and the cell survival rate of the 80 μM salvianolic acid C treatment group was 114.6% ± 3.22% (p < 0.0001). It shows that salvianolic acid C can significantly improve the survival rate of cardiomyocytes under the action of CoCl2. The results are shown in Figure 2 .

[0033] Example 3: ISO-induced pressure overload in vitro heart failure model

[0034] Using rat H9C2 cells as the research object, 5 groups were set up, namely the control group, the ISO group, the low-dose salvianolic acid C treatment group, the medium-dose salvianolic acid C treatment group, and the high-dose salvianolic acid C treatment group. The dose of ISO was 100 μM, the dose of salvianolic acid C was 20, 40, and 80 μM, and the control group was the solvent dimethyl sulfoxide (DMSO). After co-administering for 24 h, phalloidin staining was used to detect the cardiomyocyte hypertrophy of H9C2 cells.

[0035] The experimental results are specifically described as follows: As Figure 3 shown, compared with the control group, the cross-sectional area of cardiomyocytes in the ISO group became larger, indicating that ISO can cause myocardial hypertrophy, while the cross-sectional area of cardiomyocytes in the administration groups with different concentrations was significantly reduced, indicating that salvianolic acid C can reverse ISO-induced myocardial hypertrophy. The results are shown in Figure 3 .

[0036] Example 4: Intervention of salvianolic acid C in a doxorubicin-induced heart failure mouse model C57BL / 6J mice were used in the experiment. The mice were randomly divided into 6 groups: control group, DOX group, DOX + Sal C (10 mg / kg) treatment group, DOX + Sal C (20 mg / kg) treatment group, DOX + Sal C (40 mg / kg) treatment group, and DOX + positive control drug (dexrazoxane DXZ 30 mg / kg) treatment group, with 10 mice in each group. The dose of DOX was 5 mg / kg, once a week for 4 consecutive weeks, and the cumulative dose was 20 mg / kg; salvianolic acid C was administered by gavage; DOX and dexrazoxane were administered by intraperitoneal injection; the control group was replaced with 0.9% normal saline. After 4 weeks of continuous administration, the mice were subjected to cardiac ultrasound examination, the body weight of the mice was weighed, blood samples were collected, serum was prepared for biochemical detection, and the heart tissues of the mice were collected for later pathological detection.

[0037] Echocardiography showed that the left ventricular function parameters, namely ejection fraction EF and fractional shortening FS, of the mice in the DOX group decreased significantly, indicating the appearance of heart failure symptoms. After administration of salvianolic acid C, EF and FS were improved in a dose-dependent manner, indicating that salvianolic acid C has a positive effect in the prevention and treatment of heart failure. The results are shown in Figure 4 。

[0038] HE staining of heart tissue sections showed that in the DOX group, there were pathological phenomena such as disordered arrangement of cardiomyocytes, cytoplasmic vacuolization, and infiltration of inflammatory cells, which were significantly improved after administration of salvianolic acid C. The results are shown in Figure 5 。

[0039] Masson staining of heart tissue sections showed that collagen fiber deposition occurred in the DOX group, which was significantly improved after administration of salvianolic acid C. The results are shown in Figure 6 。

[0040] The contents of LDH, CK-MB, cTNI, and BNP in serum were detected by a biochemical detector. The results showed that the levels of the above indicators in the DOX group increased, while the contents of these indicators decreased significantly after administration of salvianolic acid C, indicating that salvianolic acid C can protect the heart from damage, thereby reducing the myocardial enzyme spectrum level and key heart failure indicators in heart failure mice. The results are shown in Figure 7 。

[0041] In summary, under multiple models, salvianolic acid C can improve heart failure-related indicators, increase the survival rate of cardiomyocytes, improve cardiomyocyte hypertrophy, increase the left ventricular ejection fraction and fractional shortening, and can also protect the heart tissue structure and improve the pathological changes of heart tissue caused by heart failure.

[0042] The above detailed description is a specific description of one of the feasible embodiments of the present invention. This embodiment is not intended to limit the protection scope of the present invention. Any equivalent implementation or modification without departing from the present invention shall be included within the scope of the technical solution of the present invention.

Claims

1. Use of salvianolic acid C in the preparation of a drug for preventing and treating heart failure, wherein the molecular formula of the salvianolic acid C is: C 26 H 20 O 10 , and the structural formula is as follows:

2. The application according to claim 1, wherein The drug can improve the viability of cardiomyocytes in a heart failure model induced by doxorubicin and cobalt chloride.

3. The application according to claim 1, wherein The drug can improve isoproterenol-induced myocardial hypertrophy.

4. The application according to claim 1, wherein The drug can improve the cardiac function injury caused by heart failure.

5. The application according to claim 1, wherein The drug can improve the pathological changes of heart tissue caused by heart failure.

6. The application according to claim 1, characterized in that The drug can reduce the levels of LDH, CK-MB, cTNI and BNP caused by heart failure.

7. The application according to claim 1, characterized in that, The drug is made of salvianolic acid C and pharmaceutically acceptable excipients.

8. The application according to claim 7, characterized in that The dosage form of the drug is an oral preparation.

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