Application of eupatorin in preparation of medicine for treating, preventing or relieving cerebral apoplexy

Abstract

The invention provides an application of eupatorin in preparation of a medicine for treating, preventing or relieving cerebral apoplexy, and the cerebral apoplexy is ischemic cerebral apoplexy and is mainly caused by nausea and encephaledema caused by brain barrier damage. By constructing a mouse model, it is proved that eupatorin can play an effective role in inhibiting brain barrier damage, and the eupatorin has important reference significance in clinical treatment of cerebral apoplexy and provides reference for preparing drugs for treating, preventing and relieving cerebral apoplexy.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular, relates to an application of hemiesterin in the preparation of a drug for treating, preventing or alleviating cerebral stroke. Background Art

[0002] Cerebral stroke is an acute cerebrovascular disease. It causes severe brain tissue damage due to sudden rupture of brain blood vessels or blood embolism that prevents blood from flowing into the brain, and has a high incidence and disability rate. Stroke is divided into ischemic stroke and hemorrhagic stroke, of which ischemic stroke accounts for 60% to 70% of the total number of strokes. At present, the main treatment methods are thrombolysis, anticoagulation or reducing intracranial pressure, but the efficacy is limited. Large-area malignant cerebral edema is the main factor leading to death from stroke, and its occurrence is closely related to the destruction of the blood-brain barrier. The blood-brain barrier (BBB) ​​is mainly composed of brain microvascular endothelial cells, pericytes, glial cells and extracellular matrix, among which the tight junctions between brain microvascular endothelial cells play an important role in the "barrier".

[0003] Eupatorin is a natural flavonoid compound that has been found to have good anti-cancer, anti-aging, and antioxidant activities. However, its application in the treatment of cerebral ischemic stroke has not been reported. Summary of the invention

[0004] The technical problem to be solved by the present invention is to provide an application of hemiesterin in the preparation of drugs for treating, preventing or alleviating cerebral stroke.

[0005] To solve the above problems, the technical solution adopted by the present invention is: A use of hemiesterin in preparing a medicine for treating, preventing or alleviating cerebral stroke.

[0006] As a further improvement of the present invention, the stroke is ischemic stroke.

[0007] As a further improvement of the present invention, the cerebral stroke is caused by large-area malignant cerebral edema.

[0008] As a further improvement of the present invention, the cerebral stroke is caused by the destruction of the blood-brain barrier.

[0009] As a further improvement of the present invention, the drug can be prepared into one of the following dosage forms: tablets, capsules, granules, powders, suspensions, emulsions, powders, solutions, gels, syrups, pills, tinctures, wine preparations, decoctions, lozenges, mixtures, suppositories, injections, inhalants or sprays.

[0010] The principle and beneficial effects of adopting the above technical solution are: The present invention has shown through animal model experiments that hemi-zosterone can protect the integrity of the blood-brain barrier during cerebral ischemia in model mice and reduce the area of ​​cerebral ischemia in model mice, and continuous administration of hemi-zosterone can significantly improve the survival rate of ischemic mice, which proves that hemi-zosterone has important practical significance and market value in the preparation of drugs for the treatment, prevention or relief of ischemic stroke. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 The leakage of Evans Blue in the cerebral cortex of mice in different groups in Example 1 of the present invention; wherein (A) is a small animal imaging diagram of Evans Blue dye leakage in the semi-toothed zeolite experimental group, solvent control group and sham operation group mice, and (B) is a statistical diagram of Evans Blue dye leakage in the semi-toothed zeolite experimental group, solvent control group and sham operation group mice; Figure 2 The test results of Example 2 of the present invention on the cerebral ischemic area of ​​cerebral ischemic mice by hemi-codontazolidine; wherein (A) is a comparison chart of Nissl staining of brain tissue of mice in the experimental group and the solvent control group, and (B) is a statistical chart of cerebral ischemic area of ​​cerebral ischemic mice in the experimental group and the solvent control group; Figure 3 It is a mouse survival rate curve diagram of Example 3 of the present invention. DETAILED DESCRIPTION

[0012] In order to make the purpose, technical solution and advantages of the present invention clearer, the present invention is described clearly and completely in conjunction with specific embodiments. The ischemic area and mean fluorescence intensity were measured using ImageJ software version 1.52i.

[0013] Preparation Example 1: Construction of the mouse middle cerebral artery occlusion (MCAO) model (1) Select a 21±0.5 g male mouse, anesthetize it with 2% tribromoethanol, and fix it in the supine position; disinfect the neck skin with 75% alcohol, cut the neck hair, make an incision on the midline of the neck, bluntly separate the subcutaneous tissue, and expose the right common carotid artery (CCA), internal carotid artery (ICA), and external carotid artery (ECA). During the operation, reduce the stimulation of the vagus nerve; (2) Ligate the proximal end of the CCA with a 6 / 0 suture, block the distal end of the ICA with an artery clamp, and hang a line at the proximal end of the ECA for later use; make an inverted "V"-shaped incision at the distal end of the ECA bifurcation, insert the line plug, and then ligate and cut the distal end of the ECA. Pull the ECA down and move it toward the ICA. At the same time, loosen the ICA artery clamp to allow the line plug to enter smoothly. When the line plug is inserted to a length of about 18-20 mm, resistance can be felt and it stops moving. This is the starting point of the middle cerebral artery (MCA); (3) Then, the ECA and the thread plug were ligated and fixed with a hanging thread to block the blood flow of the MCA. After 1.5 h of ischemia, the thread plug was removed, the ECA was ligated, and the CCA ligature was removed for reperfusion to obtain the MCAO group model mice; (4) Sham operation group: the surgical method was the same as that of the MCAO group, but the MCA blood flow was not blocked; (5) The surgical operation and 1.5 h ischemia were performed on a mouse temperature maintenance instrument. A rectal thermometer was inserted into the mouse and a heating pad was used to maintain the mouse body temperature within the range of 37°C ± 0.5°C.

[0014] Example 1: Effect of hemipteratin on the integrity of the blood-brain barrier in mice with cerebral ischemia S1: 30 MCAO model mice obtained in Preparation Example 1 were randomly divided into three groups. The eupatorin experimental group was injected with eupatorin (100 μM, 98%, Rhawn, Cat#R049958) via the tail vein at a dose of 0.1 mL / mouse immediately after reperfusion; the solvent control group (Solvent) and the sham operation group (sham) were injected with normal saline via the tail vein at a dose of 0.1 mL / mouse at the same time. S2. Blood-brain barrier leakage determination: Evans Blue dye was injected into all mice in the semi-toothed zearalenone experimental group, solvent control group and sham operation group through the tail vein at a dose of 2.5 mg / kg. After circulating in the mouse blood for 3 hours, 1× PBS (0.01M PBS phosphate buffer) was perfused into the heart. The leakage of Evans Blue dye in the brain parenchyma was quantified using the small animal in vivo imaging system.

[0015] Experimental results: Figure 1 A and Figure 1 As shown in B, the experimental results of the semi-toothed zeuratin experimental group showed that compared with the solvent control group, the Evans Blue leakage of mice in the semi-toothed zeuratin experimental group was reduced, and the difference was significant (*P<0.05), indicating that the administration of semi-toothed zeuratin can improve the integrity of the blood-brain barrier in mice with cerebral ischemia and has a protective effect on ischemic brain damage.

[0016] Example 2: Effect of hemiphthaloside on the cerebral ischemic area in mice with cerebral ischemia S1: 10 MCAO model mice obtained in Preparation Example 1 were randomly divided into two groups. The eupatorin experimental group (Eupatorin) was injected with eupatorin (100 μM, 98%, Rhawn, Cat#R049958) at a dose of 0.1 mL / mouse via the tail vein immediately after reperfusion; the solvent control group (Solvent) was injected with normal saline at a dose of 0.1 mL / mouse via the tail vein at the same time. S2: Nissl staining: Fresh mouse brain tissue was taken out 24 hours after injection and immediately fixed in 10% formaldehyde solution, dehydrated and embedded in a conventional manner; the thickness of paraffin sections was generally 5µm, and the mouse brain tissue sections were obtained by conventional dewaxing to water; Preparation of staining solution: prepare 1% tar violet solution (dissolve 1 g of tar violet in 100 ml of distilled water, filter after dissolution and set aside); Staining: Place the sections in the staining solution and place them in a 55±5℃ incubator for 30 minutes; Washing: Wash the sections with distilled water to remove excess staining solution; Differentiation: Place the slices in 70% alcohol for differentiation until the background is almost colorless; Dehydration: Dehydrate with 70%, 80%, 95% and 100% ethanol in sequence, 2 minutes each time; Transparency: Use xylene for transparent treatment, 10 minutes each time; Mounting: Use DPX.

[0017] Experimental results: Figure 2 A and Figure 2 As shown in B, the Nissl staining results showed that the ischemic area of ​​the model mice after administration of hemi-zearalenone was smaller than that of the solvent control group, and the difference was significant.

[0018] Example 3: Effect of continuous administration of hemipteratin on the survival of mice with cerebral ischemia 60 MCAO model mice obtained in Preparation Example 1 were selected and randomly divided into three groups; the hemiesterone experimental group: hemiesterone (100 μM) 0.1 mL / mouse was given for the first time after reperfusion, and the same dose of hemiesterone was given every day for the next 21 days, and the solvent control group and the sham operation group were given 0.1 mL / mouse of normal saline at the same time; the effects on the long-term survival rate of mice were observed and analyzed.

[0019] Determination of mouse survival rate: The survival rates of mice in the above-mentioned hemi-zearalenone experimental group, solvent control group and sham operation group were observed at 1d, 2d, 3d, 7d, 14d and 21d.

[0020] Experimental results: Figure 3 As shown, the results showed that the survival rate of model mice after administration of hemiesterin was improved to a certain extent compared with the solvent control group.

[0021] Based on the above results, it can be concluded that hemiphthaloside has a positive effect on improving the integrity of the blood-brain barrier, reducing the area of ​​cerebral ischemia and increasing the survival rate of mice with cerebral ischemia. Therefore, the application of hemiphthaloside in the preparation of drugs for the treatment, prevention or relief of ischemic stroke has important practical significance and practical value.

[0022] Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art may still modify the technical solutions described in the aforementioned embodiments, or make equivalent substitutions for some of the technical features therein; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A use of hemiphthaloside in the preparation of drugs for treating, preventing or alleviating stroke.

2. The use of the semi-dentate zeuratin according to claim 1 in the preparation of a drug for treating, preventing or alleviating stroke, characterized in that: The stroke is ischemic stroke.

3. The use of the semi-dentate zwitterion according to claim 1 in the preparation of a drug for treating, preventing or alleviating stroke, characterized in that: The stroke was caused by massive malignant cerebral edema.

4. The use of the semi-dentate zwitterion according to claim 1 in preparing a drug for treating, preventing or alleviating stroke, characterized in that: The stroke is caused by the breakdown of the blood-brain barrier.

5. The use of the semi-dentate zeuratin according to claim 1 in the preparation of a drug for treating, preventing or alleviating stroke, characterized in that: The drug can be prepared into one of the following dosage forms: tablets, capsules, granules, powders, suspensions, emulsions, powders, solutions, gels, syrups, pills, tinctures, wine preparations, decoctions, lozenges, mixtures, suppositories, injections, inhalants or sprays.

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