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 find that chrysophanol can significantly improve the cardiac ejection fraction of a mouse, shorten the fraction, the heart weight-body weight ratio and the lung weight-body weight ratio, reduce the left ventricular end diastolic volume, the left ventricular end systolic volume, the left ventricular end systolic diameter and the left ventricular end diastolic diameter, and improve the cardiac 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 present invention belongs to the technical field of medicine, and particularly relates to the application of chrysophanol in the preparation of drugs for anti-myocardial infarction injury. Background Art

[0002] With the acceleration of the aging process of the population and the influence of unhealthy lifestyles, the epidemic trend of cardiovascular disease risk factors in China is obvious, showing a trend of getting younger and growing rapidly. Moreover, the prevalence and mortality rates are on the rise, which is an important inducement for residents' deaths and seriously threatens the lives and health of residents. Research points out that myocardial infarction (MI) is based on coronary artery lesions, where the blood flow in the coronary artery suddenly decreases or is interrupted, causing severe and persistent acute ischemia in the corresponding myocardium, ultimately leading to ischemic necrosis of the myocardium, which can cause a large number of myocardial cells to die, and patients may develop arrhythmia, shock or heart failure. However, at present, the clinical treatment methods for myocardial infarction are still limited and cannot effectively block the progression 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), chemically named 1,8-dihydroxy-3-methylanthraquinone, is the main active ingredient isolated from Polygonaceae plants such as Rheum palmatum, Polygonum multiflorum and Polygonum cuspidatum, and has biological activities such as antioxidant, anti-inflammatory and antibacterial. Chrysophanol can reduce cerebral ischemia-reperfusion injury in mice and improve the survival rate of mice by inhibiting the activation of NLRP3 inflammasome. And chrysophanol also has a protective effect on renal ischemia-reperfusion injury in mice. However, its role in the pathophysiology of myocardial infarction (MI) is still poorly understood. Summary of the Invention

[0004] At present, the clinical treatment methods for myocardial infarction are still limited and cannot effectively block the progression 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 invention is to explore the preparation of its injection and its role in myocardial infarction diseases, 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] To achieve the above object, the present invention adopts the following technical solutions: In the first aspect, the present invention provides the application of chrysophanol in the preparation of drugs for anti-myocardial infarction injury. Experimental studies have proved that chrysophanol can reduce the area of myocardial infarction.

[0006] Furthermore, the drug also includes pharmaceutically acceptable excipients and additives, which can be routinely selected according to the dosage form requirements. Chrysophanol can be made into the required preparation with pharmaceutically acceptable excipients and additives by conventional methods, and the present invention does not limit this.

[0007] Furthermore, the dosage form of the drug is an oral preparation or an injection.

[0008] In a second aspect, the present invention also provides the use of chrysophanol in the preparation of a drug for improving cardiac function. Experiments have proved that chrysophanol can significantly increase the ejection fraction, shortening fraction, heart weight to body weight ratio, and lung weight to body weight ratio of mice, and reduce the left ventricular end-diastolic volume, left ventricular end-systolic volume, left ventricular end-systolic diameter, and left ventricular end-diastolic diameter, thereby improving cardiac function.

[0009] Furthermore, the drug also includes pharmaceutically acceptable excipients and additives, which can be routinely selected according to the dosage form requirements. Chrysophanol can be made into the required preparation with pharmaceutically acceptable excipients and additives by conventional methods, and the present invention does not limit this.

[0010] Furthermore, the dosage form of the drug is an oral preparation or an injection.

[0011] The beneficial effects of the present invention are as follows: The present invention for the first time discovers that chrysophanol can significantly increase the ejection fraction, shortening fraction, heart weight to body weight ratio, and lung weight to body weight ratio of mice, and reduce the left ventricular end-diastolic volume, left ventricular end-systolic volume, left ventricular end-systolic diameter, and left ventricular end-diastolic diameter, thereby improving cardiac function. Further research finds that chrysophanol can also significantly reduce the myocardial infarction area of mice, and shows a dose-dependent relationship. Therefore, chrysophanol can be used to prepare drugs for treating anti-myocardial infarction injury. Description of the Drawings

[0012] Figure 1 Results of mouse cardiac function detection; among them, (A) Flow chart of chrysophanol administration, LAD modeling, and echocardiogram detection time; (B) Detection of heart weight to body weight ratio (HW / BW) and lung weight to body weight ratio (LW / BW) of each group of mice; (C) Long-axis echocardiogram of the hearts of each group of mice; (D) Cardiac function index chart of each group of mice; *P<0.05, **P<0.01, ***P<0.001.

[0013] Figure 2 Results of Masson staining of mouse heart sections; Masson staining was used to detect the myocardial infarction area of each group of mice, and the blue color indicates the infarcted area, *P<0.05, **P<0.01, ***P<0.001.

[0014] Figure 3PSR staining results of mouse heart sections. PSR staining was used to detect the myocardial fibrosis area in each group of mice. The red color indicates the myocardial fibrosis region, *P<0.05, ***P<0.001.

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

[0016] To make the content of the present invention easier to understand, the technical solutions of the present invention will be further described below in conjunction with specific implementation manners, but the scope of the present invention is not limited.

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

[0018] Example 1 Construction of a mouse myocardial infarction model First, anesthetize the mice with isoflurane, then fix the mice on the operating table, cut open the skin of the mouse's neck to expose the trachea (for observing whether the tracheal intubation of the mouse is successful). After confirming the successful tracheal intubation, turn on the ventilator and anesthetic machine switches, and adjust the frequency to be consistent with the mouse's breathing frequency. Then, perform thoracotomy between the fourth intercostal space of the mouse to expose the heart, and use non-traumatic forceps to completely peel off the pericardium. Subsequently, find the heart ligation position and use a 7-0 nylon thread to ligate between the left ventricular midline and the interventricular groove. After ligation, observe whether the heart beat weakens and whether the myocardium below the ligation line turns white. If so, it indicates successful ligation; otherwise, re-ligation is required. Finally, suture the thoracic cavity, chest, and neck skin with nylon thread in sequence, turn off the anesthetic machine and ventilator, place the mice under an infrared heat lamp to recover, and perform the same surgical operation on the sham operation group, but only pass the thread through the heart without ligation.

[0019] Example 2 Detection of mouse cardiac function 2.1 Drug preparation Preparation of chrysophanol drug: Chrysophanol powder was purchased from Selleck. Calculate the chrysophanol administration dose, and prepare each reagent according to the ratio of 3% anhydrous EtOH + 2% DMSO + 40% PEG300 + 5% Tween-80 + 50% Saline. First, heat EtOH to a gentle boil at 95 °C in a micro-heater and then stop, add chrysophanol powder, vortex and mix well. Then, add DMSO, PEG300, Tween-80, and Saline in sequence, vortex and mix well.

[0020] 2.2 Drug administration In this study, male C57BL / 6J mice, weighing between 24 and 30 g (8 weeks old), were used to establish a mouse myocardial infarction model by combining left anterior descending coronary artery ligation (LAD) (see the construction steps of the mouse myocardial infarction model in Example 1). The experimental mice were randomly divided into five groups: ① Sham group; ② Myocardial infarction (MI) group, i.e., the model group; ③ MI + chrysophanol (0.1 mg / kg) group; ④ MI + chrysophanol (1 mg / kg) group; ⑤ MI + chrysophanol (10 mg / kg) group. The MI + chrysophanol (0.1 mg / kg) group, MI + chrysophanol (1 mg / kg) group, and MI + chrysophanol (10 mg / kg) group were intraperitoneally injected with the corresponding doses of chrysophanol, and the Sham group and the MI group were given an equal amount of 3% EtOH + 2% DMSO + 40% PEG300 + 5% Tween-80 + 50% Saline once a day. After continuous administration for 7 days, the MI group, MI + chrysophanol (0.1 mg / kg) group, MI + chrysophanol (1 mg / kg) group, and MI + chrysophanol (10 mg / kg) group underwent LAD surgery for model establishment, and the Sham group only passed the thread without ligation. Seven days after LAD modeling, echocardiography was used to detect the cardiac function of the mice, and pathological examinations were performed on the heart sections of the mice to compare the myocardial infarction areas of the mice in each group.

[0021] 2.3 Detection indexes Echocardiograms were 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 flow rate of isoflurane was adjusted to anesthetize the mice while maintaining their heart rate at 450 - 500 beats per minute. Chemical depilatory was used to remove hair from the anterior chest area. Rectal temperature probes and heating blankets were used to carefully maintain the body temperature at approximately 37.0 °C. Long-axis M-mode was used to measure systolic and diastolic ventricular diameters and wall thicknesses. The left ventricular fractional shortening (LVFS = [LVEDD - LVESD] × 100 / LVEDD) and left ventricular ejection fraction (LVEF = [LVEDV - LVESV] × 100 / LVEDV) were calculated to evaluate systolic function. Systolic and diastolic blood pressure anatomical parameters were obtained from M-mode tracings at the mid-papillary level. The results are as Figure 1 shown. Compared with the model group, chrysophanol could significantly increase the heart weight / body weight ratio (HW / BW, Figure 1 B) and lung weight / body weight ratio (LW / BW, Figure 1B) and the ejection fraction (FS) and shortening fraction (EF) of the mouse heart, and reduce the left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular end-systolic diameter (LVESD) and left ventricular end-diastolic diameter (LVEDD), improving cardiac function.

[0022] Example 3 Pathological Detection 3.1 Masson Staining After dewaxing the paraffin sections of the mouse heart to water, the sections were immersed in the mordant solution and incubated at room temperature for 8 - 12 h or placed in an incubator at 57℃ - 60℃ for 1 h for mordanting, then washed 3 times with distilled water, 3 min each time. Celestine blue staining solution was dropped and stained for 2 - 3 min, and then washed 2 times with distilled water, 10 - 15 s each time. Mayer hematoxylin staining solution was dropped and stained for 2 - 3 min, and then washed 2 times with distilled water, 10 - 15 s each time. Acidic differentiating solution was used for differentiating for several seconds, and distilled water was used to wash to terminate differentiation, and then washed with tap water for 10 min (blue return). Ponceau fuchsin staining solution was dropped and stained for 5 - 10 min, and then washed 2 times with distilled water, 10 - 15 s each time. Phosphomolybdic acid solution was used for differentiating treatment for 5 - 10 min. After pouring off the supernatant, the sections were not washed with water, and directly dropped with aniline blue staining solution and stained for 3 - 5 min. After washing off the aniline blue solution with a weak acid solution, continue to drop the weak acid solution to cover the sections and treat for 2 min. Dehydrate quickly with 95% ethanol for 3 - 5 s. Dehydrate with absolute ethanol 2 times, 5 - 10 s each time. Clear with xylene 2 times, 1 - 2 min each time. Seal with neutral gum and observe under an upright microscope. The results are as Figure 2 shown. Compared with the model group, chrysophanol can significantly reduce the myocardial infarction area of mice and shows a dose-dependence.

[0023] 3.2 PSR Staining After dewaxing the paraffin sections of the mouse heart to water, Sirius red staining solution was dropped and stained for 10 - 15 min, and the sections were quickly rinsed with distilled water to remove the excess staining solution. Start serial ethanol dehydration from 75%, clear with xylene, seal with neutral gum and observe under an upright microscope. The results are as Figure 3 shown. Compared with the model group, chrysophanol can significantly reduce the myocardial fibrosis area of mice with myocardial infarction and shows a dose-dependence.

[0024] 3.3 TTC Staining After the modeling of each group of animals was completed, the mouse hearts were perfused. Transferred in PBS solution at 0 - 4℃ and frozen in a -20℃ refrigerator for 30 min. Cut heart sections with a thickness of 500 um, put the sections into 1% red tetrazolium solution and incubate in a water bath at 37℃ in the dark for 30 min, gently shake the container every 5 min to ensure sufficient staining. The results are as Figure 4 shown. Compared with the model group, chrysophanol can significantly reduce the myocardial infarction area of mice and shows a dose-dependence.

[0025] In summary, compared with the model group, chrysophanol can significantly increase the fractional shortening (FS), ejection fraction (EF), heart weight / body weight ratio (HW / BW), and lung weight / body weight ratio (LW / BW) of mice, and decrease the left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), left ventricular end-systolic diameter (LVESD), and left ventricular end-diastolic diameter (LVEDD), thereby improving cardiac function. Further pathological examination results show that chrysophanol can significantly reduce the myocardial infarction area in mice. Moreover, the improvement effect of chrysophanol on myocardial infarction injury in mice is dose-dependent. Based on the above results, it is indicated that intraperitoneal injection of chrysophanol can effectively improve myocardial infarction injury and enhance cardiac function in mice.

[0026] The above are only the preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope of the present invention.

Claims

1. Application of chrysophanol in preparing drugs for anti-myocardial infarction injury, characterized in that: The drug active ingredient includes chrysophanol.

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 dosage form of the drug is an oral preparation or an injection.

4. Use of chrysophanol in the preparation of a drug for improving cardiac function, characterized in that: The drug active ingredient includes chrysophanol.

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 dosage form of the drug is an oral preparation or an injection.

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