CN113143914A

CN113143914A - Application of brucea javanica picrol in preparation of medicine for treating cerebral ischemic stroke

Info

Publication number: CN113143914A
Application number: CN202110416780.8A
Authority: CN
Inventors: 陆亚鹏; 王丹; 王雪婷; 朱俐
Original assignee: Nantong University
Current assignee: Nantong University
Priority date: 2021-04-19
Filing date: 2021-04-19
Publication date: 2021-07-23

Abstract

The invention belongs to the technical field of medicines, and discloses an application of brucea javanica picrol in preparing a medicine for treating ischemic stroke, so that a new medicine is provided for treating ischemic stroke. The study finds that the brucea javanica picrol can obviously improve neurobehavioral disturbance caused by cerebral ischemia reperfusion injury of rats, reduce the volume of cerebral infarction and relieve cerebral edema of cerebral ischemia rats. Therefore, the brucea javanica picrol has the potential of becoming a medicine for preventing and treating cerebral arterial thrombosis and has stronger clinical application value.

Description

Application of brucea javanica picrol in preparation of medicine for treating cerebral ischemic stroke

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to application of brucea javanica picrol in preparation of a medicine for treating ischemic stroke.

Background

The incidence of cerebral apoplexy in cardiovascular diseases is always high, and especially ischemic cerebral apoplexy is the majority. Ischemic stroke is a sudden neurological deficit due to ischemia and permanent cerebral infarction. Common causes include atherosclerotic occlusion of the aorta, thromboembolic infarction, lacunar infarction, and watershed regional infarcts caused by narrowing of the distal artery and decreased cerebral blood flow. About 1/3 stroke patients die soon after the disease, survivors lose working ability and even self-care ability due to hemiplegia, aphasia and other sequelae. Ischemic stroke is a common disease, seriously harms the health of human beings, affects the life quality of human beings and brings a heavy economic burden to families of patients.

Mechanisms that cause brain tissue damage following cerebral ischemia include edema, microvascular thrombosis, programmed cell death (apoptosis) and cell necrosis from infarction. Among them, inflammatory mediators (such as IL-1 beta, tumor necrosis factor-alpha, etc.) are involved in mediating cerebral edema and microvascular thrombosis, causing increased intracranial pressure. In addition, cellular necrosis may be caused by a number of factors, including depletion of stored ATP, electrolyte disturbances (including calcium influx), lipid peroxidation of cell membranes by free radicals, excitotoxicity, acidosis due to accumulation of lactic acid, and the like. The medicines for resisting cerebral ischemic stroke are various in types, but the curative effect of most medicines is not satisfactory, and the medicines have large toxic and side effects. Therefore, research and development of a safe and effective new drug for preventing and treating cerebral arterial thrombosis is a difficult task in front of us.

Brucea javanica is a plant widely distributed in asian areas, particularly in southern regions of china. As a Chinese medicinal material, it can be used for treating cancer, amebic dysentery, malaria, etc. Brucea javanica picrol is a quinone compound isolated from brucea javanica and is used for anticancer, anti-infection, antimalarial, antiviral, etc. Brucea javanica picrol can inhibit the proliferation of tumor cells, but has low toxicity to normal cells. At present, no literature report is found on the research and application of brucea javanica picrol in the field of prevention and treatment of ischemic stroke.

Disclosure of Invention

The invention aims to provide application of brucea javanica picrol in preparing a medicine for treating ischemic stroke, so that a new medicine is provided for treating ischemic stroke.

The invention provides an application of brucea javanica picrol in preparing a medicine for treating ischemic stroke, wherein the molecular formula of the brucea javanica picrol is C₂₆H₃₂O₁₁Molecular weight is 520.54, and molecular structural formula is:

preferably, the medicament is a pharmaceutical composition prepared by combining bruceol and a pharmaceutical excipient or a carrier.

Preferably, the pharmaceutical excipient is any one or a combination of at least two of a solvent, an emulsifier, a plasticizer, a disintegrant, a filler, a binder, a sweetener, or a lubricant.

Preferably, the dosage form of the pharmaceutical composition can be liquid preparation, granule, tablet, granule, soft capsule, sustained release agent, drop pill or injection.

Preferably, the pharmaceutical composition is in the form of injection

Preferably, the administration mode of the pharmaceutical composition can be oral, intravenous injection or intraperitoneal injection.

Preferably, the ischemic stroke is ischemic stroke in an acute stage and a repair stage.

Compared with the prior art, the invention provides the application of brucea javanica picrol in preparing the medicine for treating ischemic stroke, and the brucea javanica picrol has the following advantages and beneficial effects: the study finds that the brucea javanica picrol can obviously improve neurobehavioral disturbance caused by cerebral ischemia reperfusion injury of rats, reduce the volume of cerebral infarction and relieve cerebral edema of cerebral ischemia rats. Therefore, the brucea javanica picrol has the potential of becoming a medicine for preventing and treating cerebral arterial thrombosis and has stronger clinical application value.

Drawings

FIG. 1 is a photograph of brain slices of rats of the control group, the model group, 5mg/kg of brucinol group, and 10mg/kg of brucinol group of example 1.

Detailed Description

For a further understanding of the invention, reference will now be made to the preferred embodiments of the present invention by way of example, and it is to be understood that the description is intended to further illustrate features and advantages of the present invention and is not intended to limit the scope of the claims which follow.

All of the starting materials of the present invention, without particular limitation as to their source, may be purchased commercially or prepared according to conventional methods well known to those skilled in the art.

The rats used in the experiments of the preferred embodiment of the invention were healthy adult male Sprague-Dawley (SD) rats, clean grade, weighing 240-260g, purchased from the university of Nantong laboratory animals center.

Brucea javanica picrol used in the preferred embodiment of the invention was purchased from Sigma-Aldrich; the adopted 2,3,5-triphenyltetrazolium (TTC) powder is purchased from Shanghai Linggi Fine chemical industry Co., Ltd.

The brucea javanica picrol injection adopted in the preferred embodiment of the invention is an injection prepared by combining brucea javanica picrol and a solvent, wherein the solvent consists of 10% of DMSO, 40% of polyethylene glycol 200 and 50% of normal saline.

The term "reperfusion" in the present invention refers to restoration of blood flow supply.

Example 1

Brucea javanica picrol was tested for protection against cerebral ischemia reperfusion injury in SD (Sprague-Dawley) rats:

the 48 rats were randomly divided into 4 groups, designated as control group, model group, 5mg/kg brucea javanica alcohol group and 10mg/kg brucea javanica alcohol group, 12 per group.

Model group: a rat focal cerebral ischemia reperfusion model was prepared by Middle Cerebral Artery Occlusion (MCAO) with reference to the Longa EZ procedure and a slight modification. The average depth of the nylon thread inserted is 18.5 mm soil (from the ECA bifurcation) and 0.5mm, the nylon thread is tied and fixed, the skin is sutured, and the thread end of l cm is left outside the thread. The skin is sutured for 2h and then perfused for 24h, the neck does not need to be narcotized and cut again during perfusion, the head end of the nylon thread reaches the ICA bifurcation incision after the nylon thread is pulled up to have resistance, the blood flow is recanalized, and the homolateral Hornor and the contralateral limb movement disorder which are caused by the vascular embolism appear after the rat revives are the model success.

Control group: a control rat model was prepared by inserting nylon thread as per the model group, but at an average depth of l.0cm (from the bifurcation of ECA), and following the same procedure as the model group.

5mg/kg brucea javanica picrol group: the rat focal cerebral ischemia reperfusion model is prepared according to the method of the model group, and the rat is injected with brucea javanica picrol injection by the abdominal cavity while perfusing, wherein the dosage is 5 mg/kg.

10mg/kg brucea javanica picrol group: the rat focal cerebral ischemia reperfusion model is prepared according to the method of the model group, and the rat is injected with brucea javanica picrol injection by the abdominal cavity while perfusing, wherein the dosage is 10 mg/kg.

Neuroethological scoring

The neurobehavioral characteristics of rats in the model group, the control group, 5mg/kg bruceol group and 10mg/kg bruceol group were scored with reference to the Zea Longa 6 assay. The scoring criteria were: 0 minute: no symptoms of nerve damage; 1 minute: the contralateral anterior paw cannot be fully extended; and 2, dividing: turning to the outside; and 3, dividing: pouring towards the opposite side; and 4, dividing: the person cannot walk spontaneously and consciousness is lost; and 5, dividing: and death.

TABLE 1 neurobehavioral scores for groups of rats

Group of	Number of animals	Neuroethological scoring
			Control group	12	0±0
Model set	12	3.17±0.75
			5mg/kg brucea javanica picrol group	12	1.17±0.75***
10mg/kg brucea javanica picrol group	12	0.67±0.82***

Note: in comparison to model groups, denotes p <0.001

The neurobehavioral scoring results of the rats in each group are shown in Table 1, the animals in the control group have no neurological deficit, and the other groups have neurological deficit scores of different degrees. The 5mg/kg brucea javanica alcohol group and the 10mg/kg brucea javanica alcohol group have very significant differences (p <0.001) from the neurobehavioral scores of the model group. The brucea javanica picrol can obviously improve the neurobehavioral function of the ischemia-reperfusion rat and has dose dependence.

Cerebral infarction focus measurement

Taking 6 rats of a model group, a control group, 5mg/kg brucea javanica alcohol group and 10mg/kg brucea javanica alcohol group respectively, quickly taking brains, removing olfactory brains, cerebellum and lower brainstem, freezing for 10 minutes, making continuous forehead slices with the thickness of 2mm from the forehead to the occipital, placing the slices in 0.5% TTC solution, incubating for 30 minutes in a dark place at 37 ℃, and shaking the slices at intervals of 5 minutes. The sections were then fixed in 4% paraformaldehyde buffer and photographed after 24h, the resulting photograph being shown in FIG. 1, where the black area is normal brain tissue and the white area is the infarct area.

Inputting the pictures into a computer, calculating the infarct area by using Image J & Scion Image processing software, multiplying the sum of the infarct areas of the brain slices by the thickness (2mm) to obtain the total infarct volume, and determining the ratio of the volume of a cerebral infarct focus to the volume of the whole brain as the percentage volume of the cerebral infarct.

TABLE 2 percent volume of cerebral infarction in rats of each group

Group of	Number of animals	Percent volume of cerebral infarction (%)
			Control group	6	0±0
Model set	6	11.52±4.16
			5mg/kg brucea javanica picrol group	6	6.10±1.55***
10mg/kg brucea javanica picrol group	6	2.81±1.60***

Note: in comparison to the model group, denotes p < 0.001.

The results of the cerebral infarction percentage volume of each group of rats are shown in table 2, compared with the model group, the cerebral infarction percentage volume of the brucea javanica picrol group is obviously reduced, and the effect is most obvious when the brucea javanica picrol high dose group is used (p is less than 0.001).

Determination of brain Water content

Respectively taking 6 rats of a model group, a control group, 5mg/kg brucea javanica alcohol group and 10mg/kg brucea javanica alcohol group, rapidly cutting off the heads of the rats to take brains, and measuring the moisture content of brain tissues by a dry-wet method, wherein the specific steps are as follows: rapidly cutting off the head of a rat on an ice block to take out the brain, then placing the brain tissue taken out in a culture dish with qualitative filter paper wetted by physiological saline to prevent water from evaporating, and simultaneously rapidly removing the pia mater and the blood clot on the surface of the cerebral cortex; accurately weighing the weight of the left hemisphere and the right hemisphere of the brain respectively by using an analytical balance, then placing the hemispheres in an oven at 100 ℃ for 24 hours, and weighing again, wherein the relative water content is defined as [ wet weight-dry weight)/wet weight ] multiplied by 100%.

TABLE 3 relative Water content of rat brain in each group

Note: in comparison to the control, # # indicates p < 0.01; compared to the model group, denotes p <0.05, denotes p < 0.01.

The results of the measurement of the relative water content of the brains of the rats in each group are shown in Table 3, and the water content of the ischemic lateral brain tissue (right hemisphere) of the rats of 5mg/kg of brucea javanica alcohol group and 10mg/kg of brucea javanica alcohol group is significantly lower than that of the rats in the model group compared with the model group. The result shows that the brucea javanica picrol can obviously reduce the water content of the cerebral tissue damaged by ischemia reperfusion and reduce the edema degree of the cerebral hemisphere at the ischemic side.

While there have been shown and described what are at present considered the fundamental principles and essential features of the invention and its advantages, it will be apparent to those skilled in the art that the invention is not limited to the details of the foregoing exemplary embodiments, but is capable of other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. Application of brucea javanica picrol in preparation of medicine for treating cerebral ischemic strokeThe molecular formula of brucea javanica picrol is C₂₆H₃₂O₁₁The molecular weight is 520.54, and the molecular structural formula is shown as the following formula:

2. the use as claimed in claim 1, wherein the medicament is a pharmaceutical composition prepared by combining bruceol with a pharmaceutical excipient or carrier.

3. The use according to claim 2, wherein the pharmaceutical excipient is any one or a combination of at least two of a solvent, an emulsifier, a plasticizer, a disintegrant, a filler, a binder, a sweetener, or a lubricant.

4. The use of claim 2, wherein the pharmaceutical composition is in the form of liquid, granule, tablet, granule, capsule, sustained release preparation, drop pill or injection.

5. The use according to claim 2, wherein the pharmaceutical composition is in the form of an injection.

6. The use according to claim 2, wherein the pharmaceutical composition is administered orally, intravenously or intraperitoneally.

7. The use of claim 1, wherein the ischemic stroke is an acute or a repair ischemic stroke.