Anti-cardiotoxicity application of diosmetin

Abstract

The invention discloses application of diosmetin in preparation of a medicine for preventing and / or treating cardiotoxicity or diseases caused by cardiotoxicity.

Description

Technical Field

[0001] The present application is generally in the field of medicine, and specifically relates to the anti-cardiotoxic use of diosmylin. Background Art

[0002] The anthracycline drug doxorubicin (DOX) and its derivatives are widely used clinically as broad-spectrum cancer chemotherapy drugs, but at the same time, their severe dose-dependent cardiotoxic side effects can lead to myocardial damage, ultimately causing arrhythmias, systolic and diastolic dysfunction, dilated cardiomyopathy, and ultimately congestive heart failure. It is reported that the incidence of heart failure is as high as 26% in patients treated with DOX or DOX derivatives, and it may occur acutely during treatment or several years after the end of chemotherapy. Therefore, based on the anti-tumor effects of DOX and its derivatives, the development of drugs that protect the heart from damage is a hot topic of current research.

[0003] Current methods for preventing and reducing DOX toxicity include: changing from a one-time infusion of DOX to continuous infusion; using liposome-encapsulated DOX; using the iron chelator dexrazoxane; using renin-angiotensin (RAS) and beta-blockers; and applying traditional Chinese medicine (resveratrol, pyrrolidine), etc. However, there is currently no unified understanding of the application of traditional Chinese medicine in this field.

[0004] There is an urgent need for drugs to treat or prevent diseases caused by anti-cardiotoxicity, especially DOX-induced cardiotoxicity. Summary of the invention

[0005] One or more embodiments of the present application provide the use of diosmetin in preparing a medicament for preventing and / or treating cardiotoxicity.

[0006] In one or more embodiments, the cardiotoxicity is doxorubicin-induced cardiotoxicity.

[0007] In one or more embodiments, the diosmetin is administered simultaneously with doxorubicin.

[0008] One or more embodiments of the present application provide the use of diosmetin in preparing a medicament for preventing and / or treating diseases caused by cardiotoxicity.

[0009] In one or more embodiments, the cardiotoxicity is doxorubicin-induced cardiotoxicity.

[0010] In one or more embodiments, the diosmetin is administered simultaneously with doxorubicin.

[0011] In one or more embodiments, the disease caused by cardiotoxicity is myocardial damage, arrhythmia, systolic and / or diastolic dysfunction, dilated cardiomyopathy or congestive heart failure.

[0012] One or more embodiments of the present application provide use of a pharmaceutical composition comprising diosmetin in the preparation of a medicament for preventing and / or treating cardiotoxicity.

[0013] One or more embodiments of the present application provide use of a pharmaceutical composition comprising diosmetin in the preparation of a medicament for preventing and / or treating diseases caused by cardiotoxicity.

[0014] In one or more embodiments, the cardiotoxicity is doxorubicin-induced cardiotoxicity.

[0015] In one or more embodiments, the disease caused by cardiotoxicity is myocardial damage, arrhythmia, systolic and / or diastolic dysfunction, dilated cardiomyopathy or congestive heart failure.

[0016] In one or more embodiments, the pharmaceutical composition further comprises doxorubicin.

[0017] In one or more embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0018] In one or more embodiments, the term "treatment" and its derivatives include prophylactic treatment, therapeutic treatment, symptomatic treatment, and causal treatment.

[0019] In one or more embodiments, the term "prevention" and its derivative terms refer to the use of various clinical treatment methods to provide timely treatment to those suffering from or susceptible to certain diseases, prevent deterioration, enable early recovery from the disease, reduce the adverse effects of the disease, and prevent complications and disability.

[0020] In one or more embodiments, the term "simultaneous administration" and its derivatives include simultaneous administration, administration in close proximity to each other, and sequential administration separated in any manner. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 A two-dimensional diagram showing ultrasound detection of mouse cardiac beats.

[0022] Figure 2 Represents the experimental results of cardiac ultrasound detection in mice.

[0023] Figure 3 This is a HE staining image of mouse heart.

[0024] Figure 4 Represents the levels of CK-MB, BNP and MDA in mouse serum. DETAILED DESCRIPTION

[0025] Materials and Methods

[0026] Drugs and reagents: Dioscorea linalool (purity 98%) was purchased from Chengdu Lemeitian Pharmaceutical Technology Co., Ltd. (batch number: DST230406-018); Doxorubicin HCl (Doxorubicin HCl), Selleck Biotechnology Co., Ltd., USA; Dexamethasone; Mouse creatine kinase MB isoenzyme (CK-MB) ELISA detection kit, mouse brain natriuretic peptide / brain natriuretic peptide (BNP) ELISA detection kit, Jianglai Biological; Malondialdehyde (MDA) colorimetric test kit, Wuhan Elerite Biotechnology Co., Ltd.

[0027] Instruments: Multifunctional microplate detector (TECAN SPARK, Switzerland); Ultrasonic diagnostic system (PHILIPS); Desktop high-speed refrigerated centrifuge TGL-16K (Hunan Xiangyi Laboratory Instrument Development Co., Ltd.); Upright fluorescence microscope (LEICA, Germany).

[0028] Experimental animals and culture: Male C57BL / 6 mice (54, 8 weeks old, 20-23 g) were purchased from Henan Sikebes Biotechnology Co., Ltd., animal license number SCXK (Yu) 2020-0005. Mice were maintained under controlled conditions (22-24 ° C, 55%-60% humidity, and 12 h light-dark cycle) with free access to standard feed and water. All experimental procedures were in accordance with the "(Experimental Animal Management Regulations". All animal experimental protocols were approved by the Ethics Committee for the Care and Use of Experimental Animals of the Xinjiang Uygur Autonomous Region Institute of Pharmacology.

[0029] Experimental methods

[0030] Animal grouping and model establishment: The mice were allowed to acclimate to the environment one week before the experiment, and then randomly divided into 9 groups according to body weight, namely, control group, DOX group (also model group), DOX combined with dioscorea administration group (the experimental period was 14 days, dioscorea was administered by gavage throughout the whole process, and DOX was injected on the 8th and 12th days to establish the model; divided into 5 dosage groups: 7.5 mg / kg.d, 15 mg / kg·d, 30 mg / kg.d, 60 mg / kg·d, 120 mg / kg·d), DOX combined with positive drug group (including "dextromethamine group" (100 mg / kg·dextromethamine) and "dexamethasone group" (5 mg / kg·d dexamethasone)), with 6 mice in each group.

[0031] For the control group and the model group, corresponding volume of 0.5% CMC-Na was administered by intragastric gavage (ig) every day.

[0032] For the DOX combined with dioscorea and dexamethasone groups, dioscorea dissolved in 0.5% CMC-Na and dexamethasone were respectively administered daily for 2 weeks, and DOX was injected on the 8th and 12th days to establish the model; for the dexrazoxane group, dexrazoxane was injected into the tail vein before the injection of DOX.

[0033] The model group and the DOX combined with dioscorea lignan group were injected with 10 mg / kg DOX intraperitoneally from the second week, and the control group was injected with an equal volume of isotonic saline, and then injected for the second time after three days, with a cumulative dose of DOX of 20 mg / kg. The success of the model was detected by echocardiography on the 7th day after the first administration of DOX.

[0034] Cardiac ultrasound examination: All mice were examined by ultrasound on the 15th day of the experiment (see Figure 1 ). The mice were anesthetized using isoflurane, and the skin of the anterior chest area was prepared. After the skin preparation was completed, the mice were fixed in the supine position. A high-frequency matrix probe was selected to be perpendicular to the left chest wall and formed an angle of 10 to 30 degrees with the sternum to display the long-axis image of the left ventricle from the mitral valve orifice to the apex of the heart. Under the guidance of the long-axis image of the left ventricle, the M-mode image was displayed at the point where the left ventricular internal diameter was the largest (i.e., the level of the papillary muscle), and the left ventricular internal diameter and ventricular wall thickness were measured to analyze the left ventricular function. The measurement indicators included left ventricular ejection fraction (EF), left ventricular short-axis shortening rate (FS), left ventricular internal diameter during diastole (LVIDd), and left ventricular internal diameter during systole (LVIDs).

[0035] HE staining: All mice were killed on the 8th day after the first injection of DOX, and the hearts were removed and flushed with normal saline to ensure that the blood in the heart was washed out. The hearts were then placed in formalin solution for tissue fixation, and tissue embedding and HE staining were performed 3 days later.

[0036] Determination of creatine kinase MB isoenzyme (CK-MB): Collect animal whole blood samples, place at room temperature for 2 hours or at 4°C overnight, then centrifuge at 1000×g for 20 minutes, take the supernatant, and store the serum in a -80°C refrigerator after aliquoting. Dilute the serum according to a certain ratio, use the CK-MB kit for detection, follow the instructions, and use the microplate reader to detect the OD value at a wavelength of 450nm.

[0037] Determination of mouse brain natriuretic peptide (BNP): The animal serum was taken out from the -80°C refrigerator and tested with a BNP kit according to the instructions. The OD value was detected at a wavelength of 450 nm using an enzyme marker.

[0038] Determination of malondialdehyde (MDA): The animal serum was taken out from the -80°C refrigerator and tested with the MDA kit according to the instructions. The OD value was detected at a wavelength of 532 nm using an enzyme marker.

[0039] Statistical analysis: Data analysis was performed using Excel 2019 to create tables and enter data, and Graphpad Prism8.0 was used to draw images. All data are expressed as x±SD. When the data between the two groups conformed to normal distribution, the t-test was used, and one-way ANOVA was used for group comparison. P<0.05 was considered to be statistically significant.

[0040] Experimental Results

[0041] (1) Dioscorea linalool can improve cardiac function and alleviate DOX-induced cardiotoxicity in C57BL / 6J mice

[0042] Echocardiography was used to evaluate the damage of cardiac function in C57BL / 6J mice. Figure 1 ) The results showed that compared with the control group, the left ventricle of the mice in the model group was enlarged during systole and the ventricular wall thickness was significantly reduced, which was alleviated after the administration of DOX combined with geraniol.

[0043] After the mice in the model group were injected with DOX, the left ventricular ejection fraction (EF) and left ventricular fractional shortening (FS) ( Figure 2 A. Figure 2 B) decreased significantly (P<0.001), and the left ventricular internal diameter (LVIDs) during systole increased significantly ( Figure 2 D, P<0.001). Compared with the model group, the DOX combined with dioscorea lignans group and the DOX combined with positive drug group could improve EF and FS (P<0.05), shorten the systolic left ventricular internal diameter (LVIDs) (P<0.05), but had no significant effect on the diastolic left ventricular internal diameter (LVIDd) ( Figure 2 C). It can be seen that dianthocyanin can alleviate the impaired cardiac function of C57BL / 6J mice caused by DOX.

[0044] (2) Effects of geraniol on myocardial tissue fibers in DOX-treated C57BL / 6J mice

[0045] HE staining results showed that ( Figure 3 ), compared with the control group, the myocardial fibers in the model group were arranged in a disorderly manner and broken, and the gaps were widened.

[0046] Compared with the model group, the myocardial tissue myofibrils in the DOX combined with dioscorea lignin group were more orderly and the gaps were smaller, indicating that dioscorea lignin can significantly improve the myocardial fiber disorder caused by DOX in C57BL / 6J mice.

[0047] (3) Effects of geraniol on CK-MB, BNP and MDA in the serum of DOX-treated C57BL / 6J mice

[0048] The ELISA kit method was used to evaluate the degree of cardiac injury in C57BL / 6J mice after DOX treatment. Figure 4 A- Figure 4 As shown in C: Compared with the control group, the levels of CK-MB, BNP and MDA in the heart serum of mice in the model group were significantly increased (P<0.001); compared with the model group, the levels of CK-MB, BNP and MDA in the serum of mice in the DOX combined with positive drug group and the DOX combined with dioscorea lignin group were significantly decreased (P<0.05). This result further shows that dioscorea lignin can alleviate the cardiac toxicity of mice caused by DOX.

[0049] From the above results, it can be concluded that compared with the control group, DOX can dilate the left ventricle of mice during systole, significantly reduce the thickness of the ventricular wall, significantly reduce the left ventricular ejection fraction and the short axis shortening rate of the left ventricle, increase the left ventricular internal diameter during systole, and have no significant effect on the left ventricular internal diameter during diastole. At the same time, it can cause the myocardial tissue fibers to be scattered and broken, the gaps to be widened, and the levels of myocardial enzymes such as CK-MB, BNP and MDA in the mouse heart serum increase; compared with the model group, the DOX combined with dioscorea lignin administration group can alleviate the above phenomenon, and the degree of effect is similar to that of the positive drugs dexamethasone and dexrazoxane. The above shows that DOX can cause heart failure and myocardial damage in mice, and dioscorea lignin can alleviate the cardiotoxicity of mice caused by DOX.

[0050] The above description is only a specific implementation of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Use of dioscorea in the preparation of a medicament for preventing and / or treating cardiotoxicity.

2. The use according to claim 1, wherein the cardiotoxicity is cardiotoxicity caused by doxorubicin; preferably, the diosmylin is administered simultaneously with doxorubicin.

3. Use of dioscorea in the preparation of a medicament for preventing and / or treating diseases caused by cardiotoxicity.

4. The use according to claim 3, wherein the cardiotoxicity is cardiotoxicity caused by doxorubicin; preferably, the diosmylin is administered simultaneously with doxorubicin.

5. The use according to claim 3 or 4, wherein the disease caused by cardiotoxicity is myocardial damage, arrhythmia, systolic and / or diastolic dysfunction, dilated cardiomyopathy or congestive heart failure.

6. Use of a pharmaceutical composition comprising diosmetin in the preparation of a medicament for preventing and / or treating cardiotoxicity.

7. Use of a pharmaceutical composition comprising diosmetin in the preparation of a medicament for preventing and / or treating diseases caused by cardiotoxicity.

8. The use according to claim 6 or 7, wherein the cardiotoxicity is cardiotoxicity caused by doxorubicin.

9. The use according to claim 7, wherein the disease caused by cardiotoxicity is myocardial damage, arrhythmia, systolic and / or diastolic dysfunction, dilated cardiomyopathy or congestive heart failure.

10. The use according to claim 6 or 7, wherein the pharmaceutical composition further comprises doxorubicin; preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

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