Application of ginseng epoxyalkynol in the preparation of drugs for the prevention and treatment of heart failure

Abstract

The present invention belongs to the field of traditional Chinese medicine, and specifically relates to the application of ginseng epoxy alkynol in the preparation of drugs for the prevention and treatment of heart failure. Panaxydol (PND) has a down-regulating effect on the increase of lactate dehydrogenase (LDH), creatine kinase (CK), creatine kinase isoenzyme (CK-MB), aspartate aminotransferase (AST), and α-hydroxybutyrate dehydrogenase (HBDH) caused by heart failure. The results of cardiac ultrasound cardiography show that ginseng epoxy alkynol plays a protective role in the functional damage caused by heart failure and can improve the heart function of rats with heart failure to a certain extent. The results of HE staining show that ginseng epoxy alkynol can also protect the heart tissue structure and improve the pathological changes of heart tissue caused by heart failure. The above results suggest that the drug has good application prospects in the treatment of heart failure.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine, and specifically relates to the application of ginseng epoxyalkynol in the preparation of medicines for treating heart failure. Background Art

[0002] Heart failure (HF) is a complex clinical syndrome caused by changes in cardiac structure and function due to initial myocardial damage caused by various reasons, which ultimately leads to poor ventricular pumping function. The clinical features of HF include dyspnea, fatigue, decreased exercise tolerance, and pulmonary congestion and peripheral edema caused by fluid retention. It is a common critical illness in clinical practice and is often the main complication of all organic heart diseases of different etiologies.

[0003] Heart failure is a complex clinical syndrome and a serious stage of various heart diseases. The causes of chronic heart failure include coronary heart disease, cardiomyopathy, arrhythmia, hypertension, etc. Western medicine believes that pathological "remodeling" of the myocardium is the basic mechanism for the occurrence and development of heart failure. There are two key processes leading to the progression of heart failure. One is the occurrence of myocardial death events, such as acute myocardial infarction, myocardial damage and necrosis caused by severe myocarditis, etc.; the other is the systemic response caused by overactivation of the neuroendocrine system, in which the overexcitation of the renin-angiotensin-aldosterone system and the sympathetic nervous system plays a major role. Cutting off these two key processes is the basis for the effective prevention and treatment of heart failure and can improve the prognosis of patients.

[0004] As a polyacetylene compound, ginseng epoxy alkynol (PND) is a class of 17-carbon aliphatic long-chain compounds with two conjugated acetylenic bonds in its structural skeleton. Studies have shown that ginseng epoxy alkynol has anti-inflammatory effects, anti-platelet aggregation and inhibition of cell oxidase, but the specific application and effect of ginseng epoxy alkynol in the treatment of heart failure need further study. Summary of the invention

[0005] In view of the deficiencies in the prior art, the present invention provides the use of ginseng epoxy alkynol in the preparation of a drug for treating heart failure.

[0006] In order to achieve the purpose of the present invention, the technical solution adopted is as follows:

[0007] Application of ginseng epoxyalkynol in the preparation of drugs for preventing and treating heart failure.

[0008] Preferably, the drug can down-regulate the level of one or more of LDH, CK, CK-MB, AST and HBDH caused by heart failure.

[0009] Preferably, the drug can improve cardiac function damage caused by heart failure.

[0010] Preferably, the drug can improve pathological changes in cardiac tissue caused by heart failure.

[0011] Preferably, the drug is capable of increasing cardiac output.

[0012] Preferably, the drug is capable of increasing cardiac output.

[0013] Preferably, the drug can improve cardiac ejection fraction index.

[0014] Preferably, the medicament is capable of improving the cardiac fractional shortening index.

[0015] Preferably, the ginseng epoxy alkynol is a component extracted from ingredients including but not limited to Panax notoginseng and ginseng.

[0016] Another object of the present invention is to provide a medicine for treating heart failure, the raw materials of the medicine comprising ginseng epoxyalkynol and pharmaceutically acceptable excipients.

[0017] Preferably, the dosage form of the drug includes tablets, pills, granules, powder suspensions, capsules, suppositories, oral solutions, emulsions, injections or aerosols.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] (1) The present invention found that ginseng epoxy alkynol can effectively improve heart failure and downregulate the levels of LDH, CK, CK-MB, AST and HBDH caused by heart failure.

[0020] (2) Ginseng epoxy alkynol can improve cardiac function damage, increase cardiac output, ejection fraction and shortening fraction. Cardiac ultrasound results show that ginseng epoxy alkynol has a protective effect on cardiac function damage caused by heart failure and can improve heart failure induced by left anterior descending coronary artery ligation (LDA) in rats to a certain extent.

[0021] (3) Ginseng epoxy alkynol can also protect cardiac tissue structure and improve cardiac tissue pathological changes caused by heart failure. HE staining showed that the cell structure of the ginseng epoxy alkynol group was relatively complete, with abundant and uniform cytoplasm, normal interstitium, reduced number of cell necrosis, and reduced degree of leukocyte infiltration. Therefore, ginseng epoxy alkynol has broad application prospects in the field of prevention and treatment of heart failure.

[0022] (4) Ginseng epoxyalkynol can target the pathophysiological mechanism of heart failure and prevent the decline of cardiac function. It has a good therapeutic effect in both the early and late stages of heart failure. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 is the content of aspartate aminotransferase (AST) in the myocardial enzyme spectrum of the serum of rats in each group, Figure 1From left to right are the index content results after 3 days and 28 days of administration of ginseng epoxyalkynol (PND).

[0024] Figure 2 is the content of lactate dehydrogenase (LDH) in the myocardial enzyme spectrum of the serum of rats in each group, Figure 2 From left to right are the index content results after 3 days and 28 days of administration of ginseng epoxyalkynol (PND).

[0025] Figure 3 is the content of α-hydroxybutyrate dehydrogenase (HBDH) in the myocardial enzyme spectrum of the serum of rats in each group, Figure 3 From left to right are the index content results after 3 days and 28 days of administration of ginseng epoxyalkynol (PND).

[0026] Figure 4 is the content of creatine kinase (CK) in the myocardial enzyme spectrum of the serum of rats in each group, Figure 4 From left to right are the index content results after 3 days and 28 days of administration of ginseng epoxyalkynol (PND).

[0027] Figure 5 is the content of creatine kinase isoenzyme (CK-MB) in the myocardial enzyme spectrum of the serum of rats in each group, Figure 5 From left to right are the index content results after 3 days and 28 days of administration of ginseng epoxyalkynol (PND).

[0028] Figure 6 These are the echocardiographic test results of rats in each group.

[0029] Figure 7 The cardiac function indexes of rats in each group included ejection fraction (EF), fractional shortening (FS), cardiac output (CO) and stroke volume (SV).

[0030] Figure 8 Representative images of heart tissue sections after HE staining. DETAILED DESCRIPTION

[0031] The present invention will be further described below in conjunction with specific implementation modes.

[0032] The raw materials, equipment and reagents used in the specific implementation of the present invention are all known products, which are obtained by purchasing commercially available products. Among them, Panax notoginseng was purchased from the Wenshan medicinal material base in Yunnan, and ginseng epoxy alkynol was obtained by laboratory separation. The specific preparation method is in accordance with Example 1 of Chinese invention patent CN 117717574 A, and the ginseng epoxy alkynol HPLC was determined to be ≥95%.

[0033] Example Effects of Ginseng Cyclopentanol on Treating Heart Failure and Improving Cardiac Function

[0034] 1. Preparation of Heart Failure Rat Model

[0035] After SD rats were fed with basic feed in an SPF animal room for 3 days, the animal model was prepared: 1% sodium pentobarbital was used to perform intraperitoneal anesthesia on the rats. After waiting for 5 minutes, the rats were pinched at the tip and root of the tail to observe if they had no reaction. After ensuring that the rats were in an anesthetized state, tracheal intubation was performed and the rats were connected to a small animal ventilator for artificial ventilation. The most obvious part of the heart beat was located, the skin was cut longitudinally, the muscles were bluntly separated with elbow hemostats, the heart was fully exposed with a chest opener, and the left anterior descending branch of the heart coronary artery was permanently ligated with 5-0 medical sutures. After the ligation was completed, the chest was gently squeezed to remove the air, the ribs were sutured in an "8" shape, the epidermis was sutured, and iodine was used for disinfection. Except for no ligation, the sham operation group underwent the same operation.

[0036] Before modeling, the rats' electrocardiogram rhythm was regular, the duration of each segment was normal, and the amplitude and morphology of each wave group were normal, which was a normal electrocardiogram; 1 to 5 minutes after ligation of the left anterior descending branch of the coronary artery, the electrocardiogram showed that the ST segment was arched upward, indicating that the rat myocardial ischemia model was successfully established.

[0037] 2. Grouping and Dosing

[0038] The rats were randomly divided into 6 groups, namely, sham operation group, model group, positive drug enalapril group and ginseng epoxyalkynol low-dose, medium-dose and high-dose groups, with 9 rats in each group.

[0039] The drug administration started on the second day after the model was successfully established. The low, medium and high dose groups of ginseng epoxy alkynol were given 1mg / kg, 2mg / kg and 5mg / kg of ginseng epoxy alkynol, respectively, and the positive drug group was given 5mg / kg of enalapril maleate for four consecutive weeks. Blood was collected from the orbits of rats in each group 3d and 28d after administration, and serum was obtained by centrifugation at 3500 rpm / min for 10min, stored at -80℃, and then five myocardial enzyme spectrum tests were performed. Four weeks after administration, the rats were anesthetized and the organs were stored at -80℃ for subsequent experiments.

[0040] 3. Detection of serum myocardial enzyme indexes

[0041] In this study, biochemical detector was used to detect the levels of cardiac injury markers such as LDH, CK, and CK-MB in serum to evaluate the severity of heart failure.

[0042] Data are presented as mean ± SEM and statistically analyzed using GraphPad Prism software. All data were analyzed using independent sample t-test. # P ≤ 0.05, ## P ≤0.01, #### P≤ 0.0001; compared with the model, ** P ≤0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

[0043] The results of serum AST levels after 3d and 28d of administration are shown in Figure 1 The results of serum LDH levels after 3d and 28d of administration are shown in Figure 2 The results of serum HBDH levels after 3d and 28d of administration are shown in Figure 3 The results of serum CK levels after 3d and 28d of administration are shown in Figure 4 The results of serum CK-MB levels after 3d and 28d of administration are shown in Figure 5 . As can be seen from the figure, after 3 days of administration, there was a significant statistical difference between the sham operation group and the model group, proving that the model was successfully established. After 28 days of administration, the myocardial enzyme level of the model group increased compared with the sham operation group; and after different doses of ginseng epoxy alkynol treatment during this process, the myocardial enzyme spectrum (AST, LDH, HBDH, CK and CK-MB) levels decreased, indicating that the drug can protect the heart from damage and thus reduce the myocardial enzyme spectrum levels of rats with heart failure, proving that ginseng epoxy alkynol (PND) has a certain therapeutic effect on LDA-induced heart failure model rats.

[0044] 4. Cardiac ultrasound examination

[0045] Echocardiography was performed using the Vevo 3100 high-resolution small animal ultrasound imaging system (MX250 probe, frequency 21MHz) to evaluate cardiac function. Rats were placed in the supine position on the physiological information monitoring platform and anesthetized with isoflurane inhalation. Long-axis B-ultrasound and M-ultrasound images of the heart were obtained beside the left sternum; the probe was rotated 90 degrees, and short-axis B-ultrasound and M-ultrasound images were obtained at the level of the papillary muscle. Vevo LAB ultrasound analysis software was used to accurately measure various parameters of the left ventricle, and the average value of at least 3 consecutive cardiac cycles was taken. The main indicators included ejection fraction (EF), shortening fraction (FS), stroke volume (SV) and cardiac output (CO). When the ejection fraction (EF) of the experimental rat was less than 45% and the shortening fraction (FS) was less than 30%, it indicated that the heart failure model had been successfully established.

[0046] Data are presented as mean ± SEM, and statistical analysis was performed using GraphPad Prism software. All data were analyzed using independent sample t-test. # P ≤ 0.05, ## P ≤0.01, ####P ≤0.0001; compared with the model group, * P ≤ 0.05, ** P ≤0.01, *** P ≤ 0.001, **** P ≤ 0.0001.

[0047] like Figure 6-7 As shown in the ultrasound cardiogram, the sham group had normal heart chamber size and normal left ventricular anterior wall pulsation amplitude; however, the model group had significantly increased chamber size and significantly weakened left ventricular anterior wall pulsation amplitude; compared with the model group, after treatment with different doses of ginseng epoxy alkynol, the rat heart chamber size was significantly reduced, and the left ventricular anterior wall pulsation amplitude increased, and the effect was comparable to that of the positive drug enalapril, indicating that ginseng epoxy alkynol can improve heart morphology and function. In addition, among the cardiac function test indicators, the ejection fraction (EF) of rats with heart failure dropped below 45%, and the shortening fraction (FS) was less than 30%, indicating that their heart function was severely damaged; after giving heart failure rats different doses of ginseng epoxy alkynol, their heart function was significantly improved. After high-dose (5 mg / kg) administration of ginseng epoxyalkynol, the improvement of rat cardiac function was particularly significant, and the effect was stronger than that of the positive drug enalapril maleate, as shown by a significant increase in ejection fraction (EF) and fractional shortening (FS), as well as a significant increase in stroke volume (SV) and cardiac output (CO). The above results fully demonstrate the positive effect of preventive administration of ginseng epoxyalkynol in the treatment of heart failure.

[0048] 5. Tissue Section Preparation and HE Staining

[0049] Fresh heart tissue was fixed with 4% paraformaldehyde for 24 hours, embedded in paraffin, and the trimmed wax block was placed in a paraffin slicer to slice 3-4μm thick. The slices were dewaxed, dried, and HE stained. The slices were placed in xylene I for 20 minutes, xylene II for 20 minutes, anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, and 75% alcohol for 5 minutes, and washed with tap water. Hematoxylin staining: The slices were dyed in hematoxylin staining solution for 3-5 minutes, washed with tap water, differentiated with differentiation solution, washed with tap water, blued with blue rebate solution, and rinsed with running water. Eosin staining: The slices were dehydrated in 85% and 95% gradient alcohol for 5 minutes each, and then stained in eosin staining solution for 5 minutes. Dehydration and sealing: The slices were placed in anhydrous ethanol I for 5 minutes, anhydrous ethanol II for 5 minutes, anhydrous ethanol III for 5 minutes, xylene I for 5 minutes, and xylene II for 5 minutes to make them transparent, and sealed with neutral gum. Microscope examination, image acquisition and analysis.

[0050] Results: HE staining of heart tissues of ginseng epoxyalkynol to improve heart failure is shown in Figure 8 , the cell nucleus is blue and the cytoplasm is red in the image. Compared with the sham operation group, the cardiomyocytes in the model group showed coagulative necrosis under the microscope, the cytoplasm was strongly eosinophilic, the horizontal stripes disappeared, the myocardial tissue was disordered, the cardiomyocytes in the infarct area disappeared, the nuclei mostly disappeared and were condensed, and some leukocytes were infiltrated; compared with the model group, in the tissue sections of the ginseng epoxy alkynol administration group, the cell structure was relatively complete, the cytoplasm was rich and uniform, the interstitium was normal, the number of cell necrosis was reduced, and the degree of leukocyte infiltration was reduced. It shows that ginseng epoxy alkynol can protect the heart from tissue pathological changes caused by vascular ligation to a certain extent.

[0051] Conclusion: Ginseng epoxyalkynol can effectively improve heart failure, downregulate the elevated levels of myocardial enzymes such as LDH, CK, CK-MB, AST and HBDH caused by heart failure, improve cardiac function damage, increase ejection fraction, shortening fraction, cardiac output and stroke volume, and can also protect the heart tissue structure and improve the pathological changes of heart tissue caused by heart failure.

[0052] The above detailed description is a specific description of one feasible embodiment of the present invention. The embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or modification that does not deviate from the present invention should be included in the scope of the technical solution of the present invention.

Claims

1. Use of ginseng epoxyalkynol in the preparation of drugs for preventing and treating heart failure; the drug can improve the heart function damage caused by heart failure, and the drug can improve the pathological changes of heart tissue caused by heart failure.

2. The use according to claim 1, characterized in that: The drug can down-regulate the level of one or more of LDH, CK, CK-MB, AST and HBDH caused by heart failure.

3. The use according to claim 1, characterized in that: The drug increases cardiac output.

4. The use according to claim 1, characterized in that: The drug increases cardiac output.

5. The use according to claim 1, characterized in that: The drug can improve the cardiac ejection fraction index.

6. The use according to claim 1, characterized in that: The drug can improve the cardiac fractional shortening index.

7. The use according to claim 1, characterized in that: The dosage form of the drug is selected from tablets, pills, granules, powders, suspensions, capsules, suppositories, oral solutions, emulsions, injections or aerosols.

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