Application of chrysophanol in preparation of medicine for resisting myocardial infarction injury

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of chrysophanol in preparation of a medicine for resisting myocardial infarction injury. Tests show that chrysophanol can significantly increase the ejection fraction of the heart of a mouse, shorten the fraction and improve the heart function. Further research finds that chrysophanol can also significantly reduce the myocardial infarction area of mice, and presents dose dependence. Therefore, chrysophanol can be used for preparing medicines for treating and resisting myocardial infarction injury.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of medicine, and particularly relates to application of chrysophanol in preparation of a drug for resisting myocardial infarction damage. BACKGROUND

[0002] With the acceleration of population aging and the influence of unhealthy lifestyle, the epidemic trend of cardiovascular disease risk factors in China is obvious, and presents a low-age and rapid growth trend, and the prevalence and mortality rate are continuously rising, which is an important inducement for residents' death and seriously threatens the life and health of residents. Researches show that myocardial infarction (MI) is based on coronary artery disease, and the blood flow of coronary artery is sharply reduced or interrupted, which causes the corresponding myocardium to appear serious and persistent acute ischemia, and finally leads to ischemic necrosis of myocardium, which can cause a large number of myocardial cells to die, and the patient to have arrhythmia, shock or heart failure. However, the current clinical treatment means for myocardial infarction is still limited, and cannot effectively block the development of myocardial infarction to heart failure. Therefore, exploring the pathological mechanism of myocardial infarction will bring new targets, candidate drugs and strategies for the treatment of myocardial infarction, which has important scientific significance and clinical application value.

[0003] Chrysophanol (CHR) is a main active ingredient isolated from Rheum officinale Baill, Polygonum multiflorum Thunb and Polygonum cuspidatum Sieb et Zucc, and has biological activities such as antioxidant, anti-inflammatory and antibacterial. Chrysophanol can reduce the brain ischemia-reperfusion injury of mice by inhibiting the activation of NLRP3 inflammasome, and improve the survival rate of mice. And chrysophanol also has a protective effect on renal ischemia-reperfusion injury of mice. However, its role in the pathophysiology of myocardial infarction (MI) is still little known. SUMMARY

[0004] The current clinical treatment means for myocardial infarction is still limited, and cannot effectively block the development of myocardial infarction to heart failure. The natural active compound chrysophanol extracted from traditional Chinese medicine has various drug activities such as antibacterial, anti-inflammatory and antioxidant, but its solubility is not high. Therefore, the purpose of the present application is to explore the preparation of its injection and its role in myocardial infarction disease, which can bring new targets, candidate drugs and strategies for the treatment of myocardial infarction, and has important scientific significance and clinical application value.

[0005] In order to achieve the above purpose, the technical scheme adopted by the present application is as follows: In a first aspect, the present application provides application of chrysophanol in preparation of a drug for resisting myocardial infarction damage. It is proved by experimental research that chrysophanol can reduce the area of myocardial infarction.

[0006] Further, the medicine also comprises pharmaceutically acceptable excipients and additives, which can be routinely selected according to the requirements of the dosage form, and chrysophanol can be prepared into the required preparation by a routine method with the pharmaceutically acceptable excipients and additives, which are not limited in the present application.

[0007] Further, the dosage form of the medicine is oral preparation or injection.

[0008] In the second aspect, the present application also provides the use of chrysophanol in the preparation of a medicine for improving heart function. It is proved by experiments that chrysophanol can significantly improve the ejection fraction and fractional shortening of the heart of mice and improve the heart function.

[0009] Further, the medicine also comprises pharmaceutically acceptable excipients and additives, which can be routinely selected according to the requirements of the dosage form, and chrysophanol can be prepared into the required preparation by a routine method with the pharmaceutically acceptable excipients and additives, which are not limited in the present application.

[0010] Further, the dosage form of the medicine is oral preparation or injection.

[0011] The present application has the following beneficial effects: The present application firstly finds that chrysophanol can significantly improve the ejection fraction and fractional shortening of the heart of mice and improve the heart function. Further research finds that chrysophanol can also significantly reduce the myocardial infarction area of mice in a dose-dependent manner. Therefore, chrysophanol can be used for preparing a medicine for treating myocardial infarction damage. BRIEF DESCRIPTION OF DRAWINGS

[0012] Figure 1 Figure 1 is the results of heart function detection of mice; wherein (A) is the time flow chart of chrysophanol administration, LAD modeling and echocardiography detection; (B) is the long axis echocardiogram of the heart of mice in each group; (C) is the index chart of heart function (EF, FS) of mice in each group; *P<0.05, **P<0.01, ***P<0.001.

[0013] Figure 2 Figure 3 is the Masson staining results of the heart sections of mice; Masson staining is used to detect the myocardial infarction area of mice in each group, and the blue color represents the infarction area; *P<0.05, **P<0.01, ***P<0.001.

[0014] Figure 3 Figure 4 is the PSR staining results of the heart sections of mice; PSR staining is used to detect the myocardial fibrosis area of mice in each group, and the red color represents the myocardial fibrosis area; *P<0.05, ***P<0.001.

[0015] Figure 4The TTC staining results of the mouse heart sections, TTC staining was used to detect the myocardial infarction area of the mice in each group. White color indicates ischemic infarction area, *P<0.05, **P<0.01, ***P<0.001. DETAILED DESCRIPTION

[0016] In order to make the content of the present application easier to understand, the technical solutions of the present application will be further described below in combination with specific embodiments, but the scope of the present application is not limited thereto.

[0017] The experimental methods in the following examples are all conventional methods unless otherwise specified. The materials in the following examples can be purchased from the market unless otherwise specified.

[0018] Example 1 Construction of a mouse myocardial infarction model First, the mouse was anesthetized with isoflurane, then fixed on the operating table, the skin of the mouse neck was cut to expose the trachea (to facilitate observation of whether the tracheal intubation of the mouse was successful), after confirming the success of the tracheal intubation, the switches of the ventilator and the anesthetizing machine were turned on, and the frequency was adjusted to be consistent with the breathing frequency of the mouse; then the chest of the mouse was opened at the fourth intercostal space to expose the heart, and the whole pericardium was stripped with atraumatic forceps; then the heart ligation position was found, and a 7-0 nylon thread was used to ligate between the left ventricular midline and the interventricular groove; after ligation, it was observed whether the heart beat was weakened and the myocardium below the ligation line was whitened, if so, it indicated that the ligation was successful, otherwise it needed to be re-ligated; finally, the chest cavity, chest and neck skin were sutured in turn with nylon thread, the anesthetizing machine and the ventilator were turned off, and the mouse was placed under the infrared warm lamp for recovery. The same surgical operation was performed on the sham operation group, but only the heart was threaded without ligation.

[0019] Example 2 Detection of mouse heart function 2.1 Preparation of drugs Preparation of chrysophanol drug: Chrysophanol powder was purchased from Selleck Company, the dose of chrysophanol was calculated, and each reagent was prepared according to the ratio of 3% anhydrous EtOH + 2% DMSO + 40% PEG300 + 5% Tween-80 + 50% Saline. First, EtOH was heated to micro-boiling at 95°C in a micro-heating instrument, then chrysophanol powder was added, vortexed, mixed, and then DMSO, PEG300, Tween-80 and Saline were added in turn, vortexed and mixed.

[0020] 2.2 Drug administration C57BL / 6J male mice weighing between 24-30 g (8 weeks old) were used to construct a mouse myocardial infarction model by left anterior descending coronary artery ligation (LAD) (see the steps for constructing a mouse myocardial infarction model in Example 1). The experimental mice were randomly divided into five groups: ① Sham operation (Sham) group; ② myocardial infarction (MI) group, i.e., modeling group; ③ MI+physcion (0.1 mg / kg) group; ④ MI+physcion (1 mg / kg) group; ⑤ MI+physcion (10 mg / kg) group. The MI+physcion (0.1 mg / kg) group, MI+physcion (1 mg / kg) group, and MI+physcion (10 mg / kg) group were intraperitoneally injected with the corresponding dose of physcion, and the Sham group and MI group were given the same amount of 3% EtOH+2% DMSO+40% PEG300+5% Tween-80+50% Saline, once a day. After 7 days of continuous administration, the MI group, MI+physcion (0.1 mg / kg) group, MI+physcion (1 mg / kg) group, and MI+physcion (10 mg / kg) group were subjected to LAD modeling, and the Sham group was only threaded without ligation. After 7 days of LAD modeling, the heart function of the mice was detected by echocardiography, and the pathological examination of the mouse heart sections was performed to compare the myocardial infarction area of the mice in each group.

[0021] 2.3 Detection index An echocardiogram was obtained using an ultrasound system equipped with a high-frequency (30 MHz) linear array transducer. Echocardiography was performed 1 week after LAD surgery. Mice were anesthetized with isoflurane (3% for induction, 1-1.5% for maintenance) mixed in 1 L / min O2 through a face mask. To minimize the confounding effects of different heart rates on aortic pressure gradients and left ventricular function, the isoflurane flow was adjusted to anesthetize the mice while maintaining their heart rate at 450-500 beats per minute. Hair was removed from the anterior chest region using a chemical depilatory. Body temperature was carefully maintained close to 37.0°C using a rectal temperature probe and a heating blanket. Long-axis M-mode was used to calculate left ventricular fractional shortening (LVFS = [LVEDD-LVESD] x 100 / LVEDD) and left ventricular ejection fraction (LVEF = [LVEDV-LVESV] x 100 / LVEDV) to assess systolic function. Systolic and diastolic pressure anatomical parameters were obtained from M-mode tracings at the mid-papillary level. The results are shown in Figure 1 Compared with the modeling group, physcion can significantly improve the ejection fraction (FS) and shortening fraction (EF) of the mouse heart and improve heart function.

[0022] Example 3 Pathological examination 3.1 Masson staining After the paraffin sections of mouse heart were deparaffinated to water, the sections were immersed in the medium dyeing solution for dyeing at room temperature for 8-12 h or placed in a 57-60 °C incubator for 1 h for dyeing, and then distilled water was immersed for 3 times, 3 min each time. Azure blue staining solution was dropped for 2-3 min, and distilled water was washed for 2 times, 10-15 s each time. Mayer hematoxylin staining solution was dropped for 2-3 min, and distilled water was washed for 2 times, 10-15 s each time. Acid differentiation solution was differentiated for a few seconds, and distilled water was washed to stop differentiation, and tap water was washed for 10 min (blue return). Ponceau red staining solution was dropped for 5-10 min, and distilled water was washed for 2 times, 10-15 s each time. Phosphomolybdate solution was differentiated for 5-10 min. The upper liquid was poured off, and the sections were not washed with water, and aniline blue staining solution was directly dropped for 3-5 min. After the aniline blue solution was washed with weak acid solution, the sections were covered with weak acid solution for 2 min. 95% ethanol was rapidly dehydrated for 3-5 s. Anhydrous ethanol was dehydrated for 2 times, 5-10 s each time. Xylene was transparent for 2 times, 1-2 min each time. Neutral gum was sealed, and the sections were observed under a upright microscope. The results are shown in Figure 2 As shown in the results, compared with the model group, chrysophanol can significantly reduce the myocardial infarction area of mice in a dose-dependent manner.

[0023] 3.2 PSR staining After the paraffin sections of mouse heart were deparaffinated to water, Sirius red staining solution was dropped for 10-15 min, and the sections were rapidly washed with distilled water to remove excess dyeing solution. Starting from 75% ethanol, the sections were rapidly dehydrated, transparented with xylene, sealed with neutral gum, and then observed under a upright microscope. The results are shown in Figure 3 As shown in the results, compared with the model group, chrysophanol can significantly reduce the myocardial fibrosis area of mice in a dose-dependent manner.

[0024] 3.3 TTC staining After the modeling of each group of animals was completed, the mouse heart was perfused. The heart was transferred to a 0-4 °C PBS solution, and then frozen in a-20 °C refrigerator for 30 min. The heart section was cut, and the thickness was 500 um. The section was placed in a 1% red tetrazolium solution in a 37 °C dark water bath for 30 min, and the container was gently shaken every 5 min to ensure sufficient dyeing. The results are shown in Figure 4 As shown in the results, compared with the model group, chrysophanol can significantly reduce the myocardial infarction area of mice in a dose-dependent manner.

[0025] In summary, compared with the model group, chrysophanol can significantly improve the ejection fraction (FS) and shortening fraction (EF) of mouse heart, and improve heart function. Further pathological detection results show that chrysophanol can significantly reduce the myocardial infarction area of mice. And the improvement of chrysophanol on myocardial infarction injury of mice is in a dose-dependent manner. The above results show that intraperitoneal injection of chrysophanol can effectively improve the myocardial infarction injury of mice and enhance the heart function.

[0026] The above merely describes preferred embodiments of the present application, and any equivalent changes and modifications made within the scope of the present application should be included in the scope of the present application.

Claims

1. The application of rhein in the preparation of drugs for treating myocardial infarction injury, characterized in that: The active pharmaceutical ingredient includes rhein.

2. The application according to claim 1, characterized in that: The drug also includes pharmaceutically acceptable excipients and additives.

3. The application according to claim 1, characterized in that: The drug is available in oral or injectable form.

4. The application of rhein in the preparation of drugs for improving cardiac function, characterized in that: The active pharmaceutical ingredient includes rhein.

5. The application according to claim 4, characterized in that: The drug also includes pharmaceutically acceptable excipients and additives.

6. The application according to claim 4, characterized in that: The drug is available in oral or injectable form.

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