Application of traditional Chinese medicine composition in preparation of medicine for preventing and / or treating cerebral ischemia-reperfusion injury

Abstract

The invention provides application of a traditional Chinese medicine composition in preparation of a medicine for preventing and / or treating cerebral ischemia-reperfusion injury, and belongs to the technical field of biological medicine manufacturing. The invention discloses application of a traditional Chinese medicine composition in preparation of a medicine for preventing and / or treating cerebral ischemia-reperfusion injury. The traditional Chinese medicine composition is prepared from the following traditional Chinese medicine materials in parts by weight: 8-10 parts of fructus evodiae, 16-20 parts of fresh ginger, 8-10 parts of ginseng and 9-12 parts of Chinese dates. Experiments prove that the traditional Chinese medicine composition can obviously improve relevant indexes such as cerebral infarction volume, neurological score, weight change, survival index, histopathologic examination and nerve cell apoptosis detection in cerebral ischemia reperfusion injury. The traditional Chinese medicine composition has a pharmacodynamic effect of treating cerebral ischemia-reperfusion injury, and provides a new means for treatment of clinical indications.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of biological medicine manufacturing, and particularly relates to application of a traditional Chinese medicine composition in preparation of a medicine for preventing and / or treating cerebral ischemia-reperfusion injury. BACKGROUND

[0002] Stroke is the second leading cause of death worldwide, and its high incidence, high disability rate and high mortality rate have become a major public health challenge. Modern medical research shows that cerebral ischemia-reperfusion injury involves complex pathological mechanisms such as oxidative stress, inflammatory response and cell apoptosis. At present, clinical treatment mainly relies on intravenous thrombolysis and intravascular thrombectomy, but these methods are limited by strict time window (<4.5 hours) and high risk of bleeding, and can only benefit 5% to 10% of patients.

[0003] Wuzhuyu decoction is from Treatise on Febrile Diseases and Miscellaneous Diseases by Zhang Zhongjing of the Eastern Han Dynasty, and is the fifth prescription in the Catalogue of Ancient Classical Famous Prescriptions (First Batch) issued by the State Administration of Traditional Chinese Medicine in 2018, which is composed of four medicinal materials, i.e., Wuzhuyu, Renshen, Shengjiang and Dazao. It has been used for treating headache of jueyin, cold vomiting, cold abdominal pain and the like, and has the effects of warming middle energizer, dispelling cold, descending adverse flow and stopping vomiting. At present, there is no report about whether Wuzhuyu decoction can intervene in stroke. SUMMARY

[0004] Therefore, the purpose of the present application is to provide a new use of Wuzhuyu decoction, and application of Wuzhuyu decoction in preparation of a medicine for preventing and / or treating cerebral ischemia-reperfusion injury. The present application can significantly improve cerebral ischemia-reperfusion injury by reducing cerebral infarction volume, improving neural function, improving survival rate and inhibiting neural cell apoptosis.

[0005] The present application provides application of a traditional Chinese medicine composition in preparation of a medicine for preventing and / or treating cerebral ischemia-reperfusion injury (CIRI), wherein the traditional Chinese medicine composition is prepared from the following traditional Chinese medicine materials by weight: 8-10 parts of Wuzhuyu, 16-20 parts of Shengjiang, 8-10 parts of Renshen and 9-12 parts of Dazao.

[0006] Preferably, the CIRI includes at least one of the following injuries: neural function injury, cerebral infarction, cerebral edema, neural cell apoptosis, reduced survival rate and reduced body weight.

[0007] Preferably, the CIRI includes middle cerebral artery embolism induced by a thread plug method.

[0008] Preferably, the medicine has the pharmacodynamic effects of reducing cerebral infarction volume in CIRI, promoting recovery of neural function, improving body weight of patients, improving survival index of patients and inhibiting neural cell apoptosis.

[0009] Preferably, the traditional Chinese medicine composition is prepared from the following traditional Chinese medicine materials by weight: 9 parts of evodia rutaecarpa, 18 parts of ginger, 9 parts of ginseng, and 12 parts of jujube.

[0010] Preferably, the traditional Chinese medicine composition is prepared by extracting evodia rutaecarpa, ginger, ginseng, and jujube with water, and collecting the water extract.

[0011] Preferably, the extraction temperature is 95-100℃, the extraction is performed 2-4 times, and the extraction time is 50-70 min / time.

[0012] Preferably, after the extraction, the water extract is further subjected to vacuum concentration and drying to obtain a powder.

[0013] Preferably, the dosage form of the medicine includes the following oral dosage forms: tablets, powders, granules, capsules, decoctions, and oral liquids.

[0014] Preferably, the crude drug relative density of the traditional Chinese medicine composition in the medicine is 1.2-1.3 g / mL.

[0015] The application provides a use of a traditional Chinese medicine composition in the preparation of a medicine for preventing and / or treating CIRI, wherein the traditional Chinese medicine composition is prepared from the following traditional Chinese medicine materials by weight: 8-10 parts of evodia rutaecarpa, 16-20 parts of ginger, 8-10 parts of ginseng, and 9-12 parts of jujube. The application uses a middle cerebral artery occlusion (MCAO) mouse model constructed by a thread plug method as an experimental object, and evaluates the protective effect of evodia rutaecarpa decoction on stroke and reperfusion injury through multiple indexes and multiple dimensions. The results show that each dose group of the evodia rutaecarpa decoction significantly reduces the cerebral infarction volume and improves the neurological deficit of the mice after ischemia-reperfusion; the survival rate of the mice in the evodia rutaecarpa decoction group is increased, and the weight loss trend is alleviated, indicating that the evodia rutaecarpa decoction has a protective effect on the overall physiological state of the CIRI mice; the pathological results show that the evodia rutaecarpa decoction reduces brain tissue damage and promotes the recovery of neural function; the TUNEL experimental results show that the evodia rutaecarpa decoction can significantly reduce the neuron apoptosis induced by CIRI, and may protect the brain tissue by inhibiting the cell apoptosis pathway. The evodia rutaecarpa decoction can significantly improve CIRI in many aspects such as reducing the cerebral infarction volume, improving the neurological function, increasing the survival rate, and inhibiting neuron apoptosis. The reasonable combination of each traditional Chinese medicine component in the traditional Chinese medicine composition plays a synergistic effect, and can significantly improve the cerebral infarction volume, neurological score, body weight change, survival index, histopathological examination, and neuron apoptosis detection in the brain ischemia-reperfusion injury. The application first proves that the evodia rutaecarpa decoction has a pharmacodynamic effect against CIRI, provides laboratory data support for the expansion of the clinical indications, and expands the medical use of the classic prescription. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 Figure 4 is the result of the 7-day pharmacodynamic evaluation of Evodia Decoction on MCAO mice; A is a schematic diagram of the overall administration plan; B is the effect of Evodia Decoction on the neurological behavior score of MCAO mice; C is a representative diagram of TTC staining of mouse brain tissue; D is the effect of Evodia Decoction on the infarct volume of mouse brain tissue; the data are represented as x ± SD, n = 6; the model group is compared with the sham operation group, ### P <0.001; the administration group is compared with the model group, *** P <0.001; Figure 2 Figure 5 is the result of the effect of Evodia Decoction on the survival state of MCAO mice for 7 days; A is the survival rate curve of MCAO mice administered with Evodia Decoction for 7 days (n = 10); the sham operation group is compared with the model group, ## P <0.01; B is the body weight change curve of MCAO mice administered with Evodia Decoction for 7 days (n = 8); the administration group is compared with the model group, * P <0.05, *** P <0.001; Figure 3 Figure 6 is the result of the effect of Evodia Decoction on the brain tissue injury of MCAO mice; A is a representative diagram of H&E staining of the cortical area of mouse brain tissue (n = 3); B is a representative diagram of Nissl staining of the cortical area of mouse brain tissue (n = 3); the scale of the upper layer of the A and B diagrams is 1000 μm, the scale of the middle layer (10x) is 100 μm, and the scale of the lower layer (20x) is 50 μm; Figure 4 Figure 7 is a representative diagram of TUNEL staining of the cortical area of mouse brain tissue, and the scale is 50 μm. DETAILED DESCRIPTION

[0017] The application provides application of a traditional Chinese medicine composition in preparation of a medicine for preventing and / or treating CIRI, and the traditional Chinese medicine composition is prepared from the following traditional Chinese medicine materials in parts by weight: Evodia 8-10 parts, ginger 16-20 parts, ginseng 8-10 parts, and jujube 9-12 parts.

[0018] In the present application, the traditional Chinese medicine composition is preferably prepared from the following weight parts of traditional Chinese medicine materials: evodia 9 parts, ginger 18 parts, ginseng 9 parts and jujube 12 parts. The evodia decoction is a prescription for warming the middle and tonifying deficiency, reducing adverse and stopping vomiting, which is composed of evodia, ginseng, ginger and jujube. Among them, evodia as the monarch drug, has the effect of warming the middle and dispelling cold, and relieving pain by promoting the flow of qi; ginseng as the ministerial drug, has the function of tonifying primordial qi and invigorating the spleen and lung, which can assist in restoring the function of the spleen and stomach; ginger plays the role of an auxiliary drug in the prescription, which can harmonize the stomach and stop vomiting, warm the middle energizer, and enhance the warming effect of the whole prescription; jujube is the ministerial drug, which can tonify the middle and replenish qi, nourish blood and calm the nerves, and has the effect of regulating the spleen and stomach and nourishing yin fluid in combination with other drugs. The preparation method of the traditional Chinese medicine composition preferably extracts evodia, ginger, ginseng and jujube with water, and collects the water extract to obtain the traditional Chinese medicine composition. The extraction temperature is preferably 95-100℃; the extraction times are preferably 2-4 times, which can be 3 times; the extraction time is preferably 50-70 min / time. After extraction, the three water extracts are preferably combined, reduced pressure concentrated and dried to obtain the traditional Chinese medicine composition powder.

[0019] In the present application, the CIRI preferably includes at least one of the following injuries: nerve function injury, cerebral infarction, brain tissue edema, nerve cell apoptosis, reduced survival rate and reduced body weight. The CIRI preferably includes middle cerebral artery embolism induced by a thread plug method. In the embodiments of the present application, the mouse middle cerebral artery occlusion (MCAO) model is established as the experimental object, and the Longa neurological behavior score is used. The results show that, compared with the control group (control) mice, the model group mice show obvious neurological impairment, such as turning around and falling down, and the neurological behavior score is significantly increased. At the same time, the TTC staining results of the brain tissue of the mice show that there is no infarction in the brain tissue of the control group mice, and there is obvious infarction area in the brain tissue of the model group mice, and the average infarction area is as high as 26.47%. From the results of the body weight change and survival rate of the mice, it can be known that the survival rate of the model group mice is only 40% in 7 days, and the body weight decreases obviously, which shows that severe brain injury affects survival. The brain tissue pathology detection results show that the color of the left and right brain of the control group mice is basically the same, the cell morphology is complete, there is no edema and vacuole, the cell nucleus is full, and the nucleolus is clear; the brain tissue of the model group mice is generally colored lighter than the normal tissue, there is severe edema, a large number of vacuoles are scattered, the nucleolus is shrunk and deeply stained; compared with the control group, the model group mice have serious loss of Nissl bodies in the cerebral cortex area, neuron damage, a large number of vacuole structures appear, and most of the neurons show atrophy phenotype, weak staining and irregular distribution. This shows that cerebral ischemia reperfusion causes brain tissue injury. In addition, the TUNEL method is used to detect the apoptosis of brain tissue nerve cells, and there is no obvious TUNEL positive cell in the brain tissue of the control group mice, while the number of positive cells in the brain tissue of the MCAO group mice increases significantly, which shows that MCAO modeling can cause a large number of nerve cell apoptosis.

[0020] In the present application, the drug preferably has the efficacy of improving the following indexes: reducing the volume of cerebral infarction in CIRI, promoting the recovery of neurological function, increasing the body weight of the patient, increasing the survival index of the patient, and inhibiting the apoptosis of neural cells. In an embodiment of the present application, after the administration of Evodia decoction, the neurological impairment is significantly alleviated, and the scores of each administration group decrease, indicating that Evodia decoction can improve the neurological deficits caused by ischemic brain injury. Evodia decoction can also increase the survival rate of mice and promote the recovery of body weight, showing a significant effect of improving the survival condition of CIRI mice. The pathological detection results show that, compared with the model group, the brain tissue edema and nucleolus shrinkage of the model mice can be significantly reduced after the administration of Evodia decoction. The loss of small bodies in the cortical area is significantly reduced after the administration of Evodia decoction, the activity of neurons is greatly improved, no obvious vacuole structure is found, and the color of neurons is darker and arranged more orderly. The above indicates that Evodia decoction has a significant improvement effect on the pathological changes of the brain tissue of the model mice. The TUNEL positive cells in the brain tissue of the mice are significantly reduced after the administration of Evodia decoction, indicating that Evodia decoction has an effect of resisting the apoptosis of neural cells in MCAO mice.

[0021] In the present application, the dosage form of the drug preferably includes the following kinds of oral preparations: tablets, powders, granules, capsules, decoctions and oral liquids. The relative density of the crude drug of the traditional Chinese medicine composition in the drug is preferably 1.2-1.3 g / mL. The preparation method of the drug in the present application is not particularly limited, and the preparation method of the drug known in the art can be used.

[0022] The application of the traditional Chinese medicine composition provided in the present application in the preparation of a drug for preventing and / or treating cerebral ischemia-reperfusion injury will be described in detail below in combination with examples, but they should not be understood as limiting the protection scope of the present application.

[0023] Example 1 Preparation method of Evodia decoction Evodia 9 g, ginger 18 g, ginseng 9 g and jujube 12 g were weighed, 10 times the mass of water was added, and extraction was carried out at 100℃ for 3 times, 1 h each time. The three times of extraction solutions were combined and concentrated to dryness under reduced pressure to obtain a powder (17.5 g).

[0024] Example 2 Pharmacodynamic evaluation of Evodia decoction on cerebral ischemia-reperfusion injury 1. Establishment of middle cerebral artery occlusion (MCAO) model of mice Male C57BL / 6 mice (22 ± 3 g, 8-10 weeks) were placed in a constant temperature room (25 ± 1℃), and were allowed to freely obtain food and water in a light and dark environment for 12 h each. After one week of adaptive feeding, a modified thread plug method was used to establish a MCAO model of mice, and the specific operation was as follows: The mice were anesthetized with 1.0% sodium pentobarbital before surgery, and the mice were fixed in a supine position. After the neck was exposed and disinfected with 75% alcohol, a 1-cm incision was made in the right part of the median neck. The glands and muscles were carefully peeled off, and the common carotid artery (CCA) was separated. The internal carotid artery (ICA) and external carotid artery (ECA) were separated from the CCA, and a "Y" shape was formed. The vagus nerve adjacent to the CCA was carefully separated with an ophthalmic forceps, and a loose knot was tied at the proximal and distal ends of the CCA with 6-0 surgical suture. Then, a hemostat was clamped at the distal end, and the proximal end was completely ligated. A suitable size was cut on the CCA with a microscissors, and a silicone-coated plug was inserted. After the hemostat was loosened, the plug was inserted along the internal carotid artery until it encountered slight resistance. At this time, the depth of the plug was about 10 mm from the vascular cross. The knot was tied to fix the plug, and a penicillin-dissolved saline cotton ball was placed on the wound, and the mice were placed in an incubator to maintain the rectal temperature at 37.0±0.5℃. The time was counted from the insertion of the plug, and the plug was gently pulled out after 1.5 h to achieve post-ischemic reperfusion. The neck incision was sutured, and the mice were completely awakened from anesthesia. The neurological behavior score was performed.

[0025] 2. Animal grouping and administration All mice were divided into the following 5 groups by random number table method: (1) sham operation group (control-operated group, control): the mice were subjected to MCAO surgery but not inserted with the plug; (2) model group (Model): the mice were successfully established MCAO model; (3) low-dose Wuzhuyu decoction extract group (Wuzhuyu decoction-L, 1.5 g / kg WZYD): after the mice were established MCAO model, 1.5 g / kg Wuzhuyu decoction extract solution was administered by gavage; (4) medium-dose Wuzhuyu decoction extract group (Wuzhuyu decoction-M, 3 g / kg WZYD): after the mice were established MCAO model, 3 g / kg Wuzhuyu decoction extract solution was administered by gavage; (5) high-dose Wuzhuyu decoction extract group (Wuzhuyu decoction-H, 6 g / kg WZYD): after the mice were established MCAO model, 6 g / kg Wuzhuyu decoction extract solution was administered by gavage; the sham operation group and the model group were administered with the same volume of saline by gavage. Immediately after reperfusion, the mice were administered with solvent control or drug intervention, and the administration was continued for 7 days. The neurological score was evaluated 24 h after modeling.

[0026] Solutions for animal experiments: Prepare solutions immediately before use. Dissolve the drug in normal saline before the experiment to prepare low-dose (1.5 g / kg), medium-dose (3 g / kg), and high-dose (6 g / kg) Wuzhuyu Decoction solutions. In this example, Wuzhuyu Decoction was administered by intragastric gavage (ig).

[0027] 3. Dosage Conversion The dosage of this study was designed with reference to the clinical dosage of Wuzhuyu Decoction. Previous experiments showed that the extract powder of each dose of Wuzhuyu Decoction was 17.48g. According to the calculation of mouse dose = clinical dose (mg / d) × human body weight × conversion coefficient (9.1) / mouse body weight, the dosage of the low-dose group of Wuzhuyu Decoction extract was equal to half of the clinical dosage, which was 1.5g / kg / d; the dosage of the medium-dose group of Wuzhuyu Decoction extract was equal to the clinical dosage, which was 3g / kg / d; the dosage of the high-dose group of Wuzhuyu Decoction extract was equal to twice the clinical dosage, which was 6g / kg / d.

[0028] 4. Animal Sample Collection Seven days after MCAO modeling and drug administration, mice were anesthetized with sodium pentobarbital and quickly dissected. (1) Serum sample collection: After blood was collected from the abdominal aorta of the mouse, it was left to stand at room temperature for stratification. After the serum was separated, it was centrifuged at 3000 r / min for 10 min, the upper serum was aspirated, and the sample was transferred to a -80℃ refrigerator for testing. (2) Histopathological staining: The chest cavity was quickly opened to expose the heart. A perfusion needle was inserted from the apex of the heart, and the right atrial appendage was cut open, accompanied by venous blood outflow. 20 mL of normal saline and 20 mL of 4% paraformaldehyde were used for uniform cardiac perfusion in sequence. The mouse's limbs turned white, the blood color became lighter, the liver turned earthy yellow, and the limbs trembled, the tail was stiff, and the whole body was stiff, indicating that the cardiac perfusion was successful. Afterwards, the brain tissue was carefully removed and fixed with 10% formalin (prepared by 37%~40% formaldehyde and ultrapure water in a ratio of 1:9) for 48 h. Subsequently, the brain tissues were dehydrated in different concentrations of ethanol, permeabilized with xylene, and then embedded in paraffin.

[0029] 5. Detection indicators: Mouse neurobehavioral scoring: Longa neurobehavioral scoring was performed according to the reference (Longa EZ, Weinstein PR, Carlson S, et al. Reversible middle cerebral artery occlusion without craniectomy in rats[J]. Stroke, 1989, 20(1): 84-91.) to reflect the degree of neurological deficit. The specific evaluation criteria are as follows: 0 points: No obvious neurological deficits.

[0030] 1 point: mild neurological deficit, manifested as the mouse cannot fully stretch the contralateral forepaw, and when the mouse's tail is lifted, the mouse's body turns to the ischemic side.

[0031] 2 points: moderate neurological deficit, manifested as the mouse turns to the ischemic side when walking.

[0032] 3 points: severe neurological deficit, manifested as the mouse falls to the ischemic side due to inability to bear its own weight.

[0033] 4 points: severe neurological deficit, manifested as the mouse has no autonomous activity consciousness, or rolls like a barrel.

[0034] 5.1 TTC staining of mouse brain tissue: After 7 days of modeling, the mouse was sacrificed by cervical dislocation, and the brain tissue was removed. Then, the cerebellum, olfactory bulb and low brain stem were removed, the mouse brain was cut into 5 pieces of about 2 mm thick slices along the coronal plane, and placed in a 0.25% TTC solution in a 37°C incubator. Avoid light staining; in the meantime, use tweezers to flip the brain slices in time to make them evenly colored. After about 30 minutes of staining, the infarction area is white, the normal tissue is red, and the transition area between infarction and normal is pink. Then, the brain slices were fixed with 4% paraformaldehyde at 4°C for 24 hours, and photographed with a camera. The relative brain infarction damage rate of each mouse was calculated using IPP 6.0 software, see formula I for details: Formula I.

[0035] 5.2 Hematoxylin-eosin (HE) staining of mouse brain tissue: The wax blocks prepared in "Animal sample collection" were cut into 4 μm slices with a microtome, and HE staining and neutral resin mounting were performed. The stained sections were observed using a microscope, and the required images were collected and analyzed using an image acquisition system, with 3 samples per group.

[0036] 5.3 Nissl staining of mouse brain tissue: The wax blocks prepared in "Animal sample collection" were cut into 4 μm slices with a microtome, and the sections were stained with Nissl staining solution. The sections were differentiated with 95% alcohol until clear Nissl bodies could be observed under the microscope, and then mounted with neutral resin. The stained sections were observed using a microscope, and the required images were collected and analyzed using an image acquisition system, with 3 samples per group.

[0037] 5.4 Mouse Brain Tissue Apoptosis Detection: The wax blocks prepared in "Animal Sample Collection" were sliced ​​into thin sections using a microtome. Mouse brain tissue apoptosis was detected according to the TUNEL kit instructions. After DAPI staining, the sections were mounted in mounting medium for storage. A NIKON inverted fluorescence microscope was used for detailed observation and image recording. Each experimental group included three samples. This process aims to accurately identify and record apoptosis in brain tissue, providing an important foundation for subsequent data analysis.

[0038] 6. Experimental results: 6.1 Pharmacodynamic evaluation of Wuzhuyu decoction in MCAO mice According to the study protocol ( Figure 1 (A) Seven days after MCAO modeling, the neurobehavioral scores and statistical analysis were performed on the mice in each group. Figure 1 The mice in the control group (B) showed no neurological impairment, with scores of 0. Compared with the control group, the mice in the MCAO group showed obvious neurological impairment, such as circling and falling, and their neurobehavioral scores were significantly increased. After treatment with Wuzhuyu Decoction, the neurological impairment was significantly alleviated, and the scores of each treatment group decreased, indicating that Wuzhuyu Decoction can improve the neurological impairment caused by ischemic brain injury. Figure 1 As shown in C, Figure 1 In center C, the red area represents normal tissue, the white area represents the infarcted area, and the pink area in the middle represents the ischemic transition zone. The results showed that no infarcts were observed in the brain tissue of the control mice, while the MCAO group showed significant infarcts, with an average infarct size of 26.47%. After treatment with Evodia rutaecarpa decoction, the infarct size was significantly reduced. The average infarct size in the 3g / kg Evodia rutaecarpa decoction group dropped to 4.75%, an 82.06% decrease compared to the MCAO group, suggesting that Evodia rutaecarpa decoction can effectively reduce brain tissue damage. These results indicate that Evodia rutaecarpa decoction can significantly improve neurological function in MCAO mice and effectively reduce infarct volume, demonstrating its excellent neuroprotective effects against ischemic stroke and reperfusion injury. The 3g / kg Evodia rutaecarpa decoction group achieved the best therapeutic effect and will serve as the key dose for subsequent studies of its anti-CIRI mechanism.

[0039] 6.2 Effect of Wuzhuyu Decoction on the 7-day survival status of MCAO mice The protective effect of Wuzhuyu decoction on CIRI was further verified by evaluating the survival rate and body weight changes of MCAO mice. Figure 2As shown in Table A, the survival rate of the MCAO group was the lowest, only 40%, indicating that severe brain injury affects survival. Evodia decoction treatment improved survival, with the 3 g / kg evodia decoction group having the highest survival rate of 80%, showing the best neuroprotective effect. The survival rate of the 6 g / kg group was 50%, lower than that of the 3 g / kg evodia decoction group, suggesting that too high a dose may have adverse factors. The change in mouse body weight is shown in Table B. Figure 2 As shown in Table B, the body weight of the MCAO group of mice decreased significantly, which may be due to metabolic disorders and reduced food intake caused by ischemic injury. Evodia decoction treatment promoted weight recovery, with the 3 g / kg evodia decoction group having the best recovery effect, indicating that this dose can effectively improve metabolic abnormalities caused by brain injury and improve the quality of life. Evodia decoction showed a significant effect on improving the survival status of CIRI mice by increasing survival rate and promoting weight recovery. The 3 g / kg evodia decoction group had the best effect, which was the optimal treatment dose, and subsequent experiments will further explore its anti-CIRI mechanism around this dose.

[0040] 6.3 Effect of Evodia Decoction on the Pathological Morphological Changes of MCAO Mouse Brain Tissue HE and Nissl staining were used to observe the effect of evodia decoction on the pathological morphological changes of MCAO mouse brain tissue. The results are shown in Table C. Figure 3 As shown in Table A, the color of the left and right brain of the control group of mice remained basically the same, the cell morphology was complete, there was no edema and vacuole, the nucleus was full, and the nucleolus was clear; the overall color of the infarct area of the brain tissue of the model group of mice was lighter than that of the normal tissue, there was severe edema, a large number of vacuoles were scattered, the nucleolus was shriveled and deeply stained; compared with the model group, evodia decoction administration could significantly reduce the brain tissue edema and nucleolus shrinkage of MCAO mice. Figure 3 As shown in Table B, when observing the effect of evodia decoction on the Nissl bodies of MCAO mice, it was found that compared with the control group, the Nissl bodies in the cerebral cortex of the model group of mice were severely lost, the neurons were damaged, a large number of vacuole structures appeared, and most of the neurons showed atrophy phenotype, with weak staining and irregular distribution. After evodia decoction administration, the loss of Nissl bodies in the cortex was significantly reduced, the neuron activity was greatly improved, no obvious vacuole structure was found, and the color of the neurons was darker and the arrangement was more orderly. The above indicates that evodia decoction has a significant improvement effect on the pathological changes of MCAO mouse brain tissue.

[0041] 6.4 Effect of Evodia Decoction on MCAO Mouse Brain Tissue Neural Cell Apoptosis During the process of ischemic stroke, neural cell apoptosis occurs, which is closely related to the severity of the disease. Therefore, TUNEL method was further used to detect the effect of evodia decoction on MCAO mouse brain tissue neural cell apoptosis, and the results are shown in Table D. Figure 4The control group mice brain tissue did not find obvious TUNEL positive cells, while the MCAO group mice brain tissue positive cell number increased significantly, which showed that MCAO modeling can cause a large number of neural cell apoptosis; and the TUNEL positive cells of mice brain tissue were significantly reduced after the administration of Evodia decoction, which showed that Evodia decoction had the effect of resisting MCAO mice neural cell apoptosis.

[0042] The above only describes the preferred embodiments of the present application, and it should be noted that those skilled in the art can make several improvements and refinements without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. Use of a Chinese medicine composition in the preparation of a medicament for preventing and / or treating cerebral ischemia-reperfusion injury, characterized in that: The traditional Chinese medicine composition is prepared from the following traditional Chinese medicine materials in parts by weight: 8-10 parts of Evodia rutaecarpa, 16-20 parts of ginger, 8-10 parts of ginseng and 9-12 parts of jujube.

2. The application according to claim 1, characterized in that The cerebral ischemia-reperfusion injury includes at least one of the following injuries: neurological function damage, cerebral infarction, brain tissue edema, nerve cell apoptosis, decreased survival rate and weight loss.

3. The application according to claim 1, characterized in that The cerebral ischemia-reperfusion injury includes middle cerebral artery embolism induced by suture embolism.

4. The application according to claim 1, characterized in that The drug has the efficacy of improving the following indicators: reducing the volume of cerebral infarction in cerebral ischemia-reperfusion injury, promoting nerve function recovery, increasing patient weight, improving patient survival index and inhibiting nerve cell apoptosis.

5. The application according to claim 1, characterized in that: The traditional Chinese medicine composition is prepared by including the following traditional Chinese medicine materials in parts by weight: 9 parts of Evodia rutaecarpa, 18 parts of ginger, 9 parts of ginseng and 9 parts of jujube.

6. The application according to claim 5, characterized in that The preparation method of the traditional Chinese medicine composition comprises extracting evodia rutaecarpa, ginger, ginseng and jujube with water, and collecting the water extract.

7. The application according to claim 6, characterized in that The extraction temperature is 95-100° C.; the number of extractions is 2-4 times; and the extraction time is 50-70 min / time.

8. The use according to claim 6, characterized in that After the extraction, the method further comprises concentrating and drying the water extract under reduced pressure to obtain powder.

9. The use according to any one of claims 1 to 8, characterized in that: The dosage forms of the drug include the following types of oral preparations: tablets, powders, granules, capsules, decoctions and oral liquids.

10. The use according to claim 9, characterized in that: The relative density of the crude drugs of the traditional Chinese medicine composition in the medicine is 1.2-1.3 g / mL.

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