Use of a traditional Chinese medicine composition in the preparation of a drug for preventing and / or treating cerebral ischemia-reperfusion injury

Abstract

The application 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 application of the traditional Chinese medicine composition in preparation of the medicine for preventing and / or treating cerebral ischemia-reperfusion injury is prepared from traditional Chinese medicine materials, wherein the traditional Chinese medicine materials are prepared from evodia rutaecarpa 8-10 parts, raw ginger 16-20 parts, ginseng 8-10 parts and jujube 9-12 parts by weight. The experiment proves that the traditional Chinese medicine composition can obviously improve the cerebral infarction volume, neurological score, body weight change, survival index, histopathological examination and neural cell apoptosis detection and other related indexes in cerebral ischemia-reperfusion injury. The traditional Chinese medicine composition has the pharmacodynamic effect of treating cerebral ischemia-reperfusion injury, and provides a new means for treating the clinical indications.

Description

Technical Field

[0001] This invention belongs to the field of biomedical manufacturing technology, specifically relating to the application of a traditional Chinese medicine composition in the preparation of drugs for the prevention and / or treatment of cerebral ischemia-reperfusion injury. Background Technology

[0002] Stroke, the second leading cause of death worldwide, poses a significant public health challenge due to its high incidence, disability rate, and mortality. Modern medical research indicates that cerebral ischemia-reperfusion injury involves complex pathological mechanisms such as oxidative stress, inflammatory responses, and apoptosis. Currently, clinical treatment mainly relies on intravenous thrombolysis and endovascular thrombectomy for vascular recanalization. However, these methods are limited by a strict time window (<4.5 hours) and a high risk of bleeding, benefiting only 5%–10% of patients.

[0003] Wu Zhu Yu Tang (Evodia Decoction) originates from Zhang Zhongjing's *Treatise on Febrile and Miscellaneous Diseases* (Shanghan Zabing Lun) of the Eastern Han Dynasty. It is the fifth prescription in the *Catalogue of Famous Ancient Prescriptions (First Batch)* published by the State Administration of Traditional Chinese Medicine in 2018. It consists of four herbs: Evodia rutaecarpa, ginseng, ginger, and jujube. Historically, it has been used to treat symptoms such as Jueyin headache, vomiting due to cold, and abdominal pain due to deficiency and cold, possessing the effects of warming the middle jiao (spleen and stomach), dispelling cold, and relieving nausea and vomiting. Currently, there are no reports on whether Wu Zhu Yu Tang can intervene in stroke. Summary of the Invention

[0004] In view of this, the purpose of this invention is to provide a new use of Evodia rutaecarpa decoction in the preparation of drugs for the prevention and / or treatment of cerebral ischemia-reperfusion injury. This invention utilizes Evodia rutaecarpa decoction to significantly improve cerebral ischemia-reperfusion injury in many ways, such as reducing cerebral infarction volume, improving neurological function, increasing survival rate, and inhibiting neuronal apoptosis.

[0005] This invention provides the application of a traditional Chinese medicine composition in the preparation of a drug for the prevention and / or treatment of cerebral ischemia-reperfusion injury (CIRI). The traditional Chinese medicine composition is prepared by comprising 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.

[0006] Preferably, the CIRI includes at least one of the following injuries: neurological function impairment, cerebral infarction, cerebral edema, neuronal apoptosis, reduced survival rate, and weight loss.

[0007] Preferably, the CIRI includes suture-induced middle cerebral artery embolization.

[0008] Preferably, the drug has the efficacy of improving the following indicators: reducing the infarct volume in CIRI, promoting the recovery of neurological function, increasing patient weight, improving patient survival index, and inhibiting neuronal apoptosis.

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

[0010] Preferably, the method for preparing the traditional Chinese medicine composition involves extracting Evodia rutaecarpa, ginger, ginseng, and jujube with water and collecting the aqueous extract.

[0011] Preferably, the extraction temperature is 95~100℃; the number of extractions is 2~4; and the extraction time is 50~70min / extraction.

[0012] Preferably, after extraction, the aqueous extract is further concentrated and dried under reduced pressure to obtain a powder.

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

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

[0015] This invention provides the application of a traditional Chinese medicine composition in the preparation of a drug for the prevention and / or treatment of CIRI (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 Evodia rutaecarpa, 16-20 parts of fresh ginger, 8-10 parts of ginseng, and 9-12 parts of jujube. Using a mouse model of middle cerebral artery occlusion (MCAO) constructed using the suture occlusion method as the experimental subject, this invention evaluates the protective effect of Evodia rutaecarpa decoction on stroke and reperfusion injury through multiple indicators and dimensions. The results show that all dose groups of Evodia rutaecarpa decoction significantly reduced the infarct volume and improved neurological deficits in mice after ischemia-reperfusion. The survival rate of mice in the Evodia rutaecarpa decoction group increased, and the trend of weight loss was alleviated, indicating that it has a protective effect on the overall physiological state of CIRI mice. Pathological results show that Evodia rutaecarpa decoction reduces brain tissue damage and promotes the recovery of neurological function. TUNEL assay results show that Evodia rutaecarpa decoction can significantly reduce CIRI-induced neuronal apoptosis, possibly protecting brain tissue by inhibiting the apoptosis pathway. Wu Zhu Yu Tang can significantly improve cerebral ischemia-reperfusion injury (CIRI) by reducing infarct volume, improving neurological function, increasing survival rate, and inhibiting neuronal apoptosis. The rational combination of the various herbal components in the formula exerts a synergistic effect, significantly improving infarct volume, neurological scores, weight changes, survival index, histopathological examination, and neuronal apoptosis detection in patients with cerebral ischemia-reperfusion injury. This invention is the first to demonstrate the anti-CIRI efficacy of Wu Zhu Yu Tang, providing laboratory data support for expanding its clinical indications and broadening the medicinal applications of this classic formula. Attached Figure Description

[0016] Figure 1 The following are the results of the 7-day efficacy evaluation of Evodia rutaecarpa decoction in MCAO mice; A is a schematic diagram of the overall dosing regimen; B is the effect of Evodia rutaecarpa decoction on the neurological behavioral scores of MCAO mice; C is a representative image of TTC staining in mouse brain tissue; D is the effect of Evodia rutaecarpa decoction on the infarct volume of mouse brain tissue; all data are expressed as x ± SD, n = 6; compared with the sham-operated group, ### P <0.001; compared with the model group, the drug-treated group, *** P <0.001;

[0017] Figure 2 The results show the effect of Evodia rutaecarpa decoction on the 7-day survival status of MCAO mice; A is the 7-day survival rate curve of MCAO mice treated with Evodia rutaecarpa decoction (n = 10), comparing the sham-operated group with the model group. ## P <0.01; B is the body weight change curve of MCAO mice treated with Evodia rutaecarpa decoction for 7 days (n = 8); compared with the model group, the treatment group, * P <0.05, *** P <0.001;

[0018] Figure 3 The effect of Evodia rutaecarpa decoction on brain tissue damage in MCAO mice. Results: A is a representative map of H&E staining in the cortical region of mouse brain tissue (n = 3); B is a representative map of Nissl staining in the cortical region of mouse brain tissue (n = 3). The scale bar of the upper map in A and B is 1000 μm, the scale bar of the middle map (10×) is 100 μm, and the scale bar of the lower map (20×) is 50 μm.

[0019] Figure 4 This is a representative image of TUNEL staining in the cortical region of mouse brain tissue. The scale bar in the image is 50 μm. Detailed Implementation

[0020] This invention provides the application of a traditional Chinese medicine composition in the preparation of a medicament for the prevention and / or treatment of CIRI. The traditional Chinese medicine composition is prepared by comprising 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.

[0021] In this invention, the traditional Chinese medicine composition is preferably prepared by comprising the following parts by weight of traditional Chinese medicine materials: 9 parts of Evodia rutaecarpa, 18 parts of ginger, 9 parts of ginseng, and 12 parts of jujube. The Evodia rutaecarpa decoction is a formula for warming the middle jiao, tonifying deficiency, and relieving nausea and vomiting, composed of Evodia rutaecarpa, ginseng, ginger, and jujube. Evodia rutaecarpa, as the principal herb, has the effects of warming the middle jiao, dispelling cold, and relieving pain; ginseng, as the assistant herb, has the functions of greatly tonifying vital energy, strengthening the spleen and lungs, and can assist in restoring spleen and stomach function; ginger plays an adjuvant role in the formula, harmonizing the stomach, relieving nausea, warming the middle jiao, and enhancing the overall warming effect of the formula; jujube, as the guiding herb, can tonify the middle jiao, replenish qi, nourish blood, and calm the mind, and together with other herbs, it can harmonize the spleen and stomach and nourish yin fluids. The preferred method for preparing the traditional Chinese medicine composition is to extract Evodia rutaecarpa, ginger, ginseng, and jujube with water, and collect the aqueous extract as the traditional Chinese medicine composition. The extraction temperature is preferably 95-100℃; the number of extractions is preferably 2-4 times, and can be 3 times; the extraction time is preferably 50-70 min / extraction. After extraction, the extracts are preferably combined, concentrated under reduced pressure, and dried to obtain a traditional Chinese medicine composition powder.

[0022] In this invention, the CIRI preferably includes at least one of the following injuries: neurological function impairment, cerebral infarction, cerebral edema, neuronal apoptosis, decreased survival rate, and weight loss. The CIRI preferably includes middle cerebral artery embolization induced by suture occlusion. In this embodiment of the invention, a mouse middle cerebral artery embolization (MCAO) model was used as the experimental subject. Longa neurobehavioral scoring showed that, compared with the control group mice, the model group mice exhibited significant neurological deficits, such as circling and tipping, and their neurobehavioral scores were significantly elevated. Simultaneously, TTC staining of mouse brain tissue showed that no infarct foci were observed in the brain tissue of the control group mice, while the model group mice showed obvious infarct areas, with an average infarct area as high as 26.47%. Results on mouse weight changes and survival rate showed that the 7-day survival rate of the model group mice was only 40%, and their weight decreased significantly, indicating that severe brain injury affects survival. Pathological examination of brain tissue revealed that in the control group, the left and right hemispheres of the brain maintained essentially the same color, with intact cell morphology, no edema or vacuoles, plump nuclei, and clearly defined nucleoli. In the model group, the infarcted area of ​​the brain tissue was lighter in color than normal tissue, exhibiting severe edema, numerous scattered vacuoles, and shrunken, deeply stained nucleoli. Compared to the control group, the model group showed significant loss of Nissl bodies in the cerebral cortex, neuronal damage, and numerous vacuolated structures. Most neurons also exhibited an atrophic phenotype, with weak staining and irregular distribution. This indicates that cerebral ischemia-reperfusion injury causes brain tissue damage. Furthermore, TUNEL assay was used to detect neuronal apoptosis in the brain tissue. No significant TUNEL-positive cells were observed in the brain tissue of the control group, while the number of positive cells was significantly increased in the MCAO group, indicating that MCAO modeling induces extensive neuronal apoptosis.

[0023] In this invention, the drug preferably has the following effects: reducing the infarct volume in CIRI, promoting neurological function recovery, increasing patient weight, improving patient survival index, and inhibiting neuronal apoptosis. In one embodiment of this invention, after administration of Evodia rutaecarpa decoction, neurological function damage was significantly alleviated, and the scores of each treatment group decreased, indicating that Evodia rutaecarpa decoction can improve neurological function defects caused by ischemic brain injury. Evodia rutaecarpa decoction can also improve the survival rate of mice and promote weight recovery, demonstrating a significant effect on improving the survival status of CIRI mice. Pathological examination results showed that, compared with the model group, administration of Evodia rutaecarpa decoction significantly reduced cerebral edema and nucleolar shrinkage in model mice. After administration of Evodia rutaecarpa decoction, the loss of cortical bodies was significantly reduced, neuronal activity was greatly improved, no obvious vacuolar structures were found, and neurons were darker in color and more orderly arranged. The above indicates that Evodia rutaecarpa decoction has a significant ameliorative effect on the pathological changes of brain tissue in model mice. After administration of Evodia rutaecarpa decoction, the number of TUNEL-positive cells in mouse brain tissue was significantly reduced, indicating that Evodia rutaecarpa decoction has an anti-apoptotic effect on neuronal cells in MCAO mice.

[0024] In this invention, the dosage form of the drug preferably includes the following types of oral preparations: tablets, powders, granules, capsules, decoctions, and oral liquids. The relative density of the crude herbal composition in the drug is preferably 1.2~1.3 g / mL. This invention does not impose any particular limitation on the preparation method of the drug; any drug preparation method well known in the art can be used.

[0025] The following detailed description, in conjunction with embodiments, illustrates the application of a traditional Chinese medicine composition provided by the present invention in the preparation of a medicament for the prevention and / or treatment of cerebral ischemia-reperfusion injury. However, these descriptions should not be construed as limiting the scope of protection of the present invention.

[0026] Example 1

[0027] Preparation method of Evodia Decoction

[0028] Weigh out 9g of Evodia rutaecarpa, 18g of ginger, 9g of ginseng and 12g of jujube, add 10 times the weight of water and extract three times at 100℃ for 1 hour each time. Combine the three extracts and concentrate under reduced pressure to dry into powder (17.5g).

[0029] Example 2

[0030] Efficacy evaluation of Evodia Decoction for cerebral ischemia-reperfusion injury

[0031] 1. Establishment of a mouse model of middle cerebral artery occlusion (MCAO)

[0032] Male C57BL / 6 mice (22 ± 3 g, 8–10 weeks old) were placed in a constant temperature room (25 ± 1℃) and given free access to food and water in 12-hour light and 12-hour dark environments. After one week of acclimatization, a mouse MCAO model was established using a modified suture method, as follows:

[0033] Before surgery, mice were anesthetized with 1.0% sodium pentobarbital. The mice were fixed in a supine position, and the neck was disinfected with 75% alcohol after exposure. A 1cm incision was made in the middle of the neck on the right side. The glands and muscles were carefully dissected to separate the common carotid artery (CCA). The carotid artery was slowly separated towards the distal end, and the internal carotid artery (ICA) and external carotid artery (ECA) connected to it could be seen. The three formed a "Y". The vagus nerve adjacent to the internal carotid artery (CCA) was carefully isolated using ophthalmic forceps. A loose knot was tied at both the proximal and distal ends of the CCA using 6-0 surgical sutures. A hemostatic clip was then applied to the distal end, and the proximal end was thoroughly ligated. A suitable incision was made in the CCA using microsurgical scissors, and a silicone-coated embolic suture was inserted. After releasing the hemostatic clip, the embolic suture was inserted along the direction of the internal carotid artery until slight resistance was encountered. At this point, the embolic suture had penetrated to a depth of approximately 10 mm from the point of vascular crossing. The ligation was tightened to secure the embolic suture. A cotton ball soaked in penicillin-dissolved saline was applied to the wound, and the mouse was placed in an incubator to maintain a rectal temperature of 37.0 ± 0.5 °C. Timing was maintained from 1.5 h after embolic suture insertion. The embolic suture was gently removed after 1.5 h to achieve ischemia-reperfusion, and the neck incision was sutured. Neurological behavioral assessments were performed after the mouse fully recovered from anesthesia.

[0034] 2. Animal grouping and administration

[0035] Using a random number table method, all mice were divided into the following 5 groups: (1) Sham-operated group (control): Mice underwent MCAO surgery but no suture was inserted; (2) Model group: Mice successfully established an MCAO model; (3) Low-dose Evodia rutaecarpa extract group (Evodia rutaecarpa-L, 1.5 g / kg WZYD): After establishing an MCAO model, mice were given 1.5 g / kg Evodia rutaecarpa extract solution by gavage; (4) Medium-dose Evodia rutaecarpa extract group (Evodia rutaecarpa-M,L, 3 g / kg WZYD): After establishing an MCAO model, mice were given 3 g / kg Evodia rutaecarpa extract solution by gavage; (5) High-dose Evodia rutaecarpa extract group (Evodia rutaecarpa-H, 6 g / kg WZYD): After establishing an MCAO model, mice were given 6 g / kg Evodia rutaecarpa extract solution by gavage; The sham-operated group and the model group were given the same volume of physiological saline by gavage. After re-gavage, mice were immediately given solvent control or drug intervention for 7 consecutive days. Neurological assessment was performed 24 hours after modeling.

[0036] Animal experimental solutions: Prepare fresh before use. Dissolve the drug in physiological saline before the experiment to prepare low-dose (1.5g / kg), medium-dose (3g / kg), and high-dose (6g / kg) Wu Zhu Yu Tang solutions. In this example, Wu Zhu Yu Tang is administered via intragastric (ig).

[0037] 3. Dosage Conversion

[0038] This study designed the dosage based on the clinical dosage of Evodia rutaecarpa decoction. Previous experiments showed that each dose of Evodia rutaecarpa decoction extract powder was 17.48g. The dosage for mice was calculated as follows: mouse dosage = clinical dosage (mg / d) × human body weight × conversion factor (9.1) / mouse body weight. The dosage for the low-dose group of Evodia rutaecarpa decoction extract was half the clinical dosage, which was 1.5g / kg / d; the dosage for the medium-dose group was equal to the clinical dosage, which was 3g / kg / d; and the dosage for the high-dose group was twice the clinical dosage, which was 6g / kg / d.

[0039] 4. Animal sample collection

[0040] Seven days after MCAO modeling and administration of the drug, mice were anesthetized with sodium pentobarbital and dissected rapidly. (1) Serum sample collection: Blood was collected from the abdominal aorta of the mice and allowed to stand at room temperature for separation. After the serum separated, it was centrifuged at 3000 r / min for 10 min, and the upper serum was collected and transferred to a -80℃ refrigerator for testing. (2) Histopathological staining: The thoracic 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, with venous blood flowing out. The heart was perfused at a constant rate with 20 mL of physiological saline and 20 mL of 4% paraformaldehyde. It was observed that the limbs of the mice turned white, the blood color became lighter, the liver was yellowish-brown, and tremors of the limbs, stiff tail and rigidity of the whole body appeared, indicating that the heart perfusion was successful. Afterwards, the brain tissue was carefully removed and fixed with 10% formalin (prepared by mixing 37%~40% formaldehyde and ultrapure water in a ratio of 1:9) for 48 h. Subsequently, the brain tissue was dehydrated in ethanol of different concentrations, permeated with xylene, and then embedded in paraffin.

[0041] 5. Testing indicators:

[0042] Mouse neurobehavioral scoring: Longa's neurobehavioral score was calculated based on 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:

[0043] 0 points: No obvious neurological deficits.

[0044] 1 point: Mild neurological deficit, manifested as the mouse's inability to fully extend its contralateral forepaw, and when the mouse's tail is lifted into the air, the mouse's body turns to the ischemic contralateral side.

[0045] 2 points: Moderate neurological deficit, manifested as the mouse circling towards the ischemic side when walking.

[0046] 3 points: Severe neurological deficits, manifested as the mouse tilting to the ischemic side because it cannot support its own weight.

[0047] 4 points: Severe neurological deficits, manifested as mice lacking voluntary activity or rolling in a cylindrical manner.

[0048] 5.1 TTC Staining of Mouse Brain Tissue: Seven days after mouse modeling, mice were euthanized by cervical vertebrae, and brain tissue was extracted. Subsequently, the cerebellum, olfactory bulb, and lower brainstem were removed. The mouse brain was cut into five thin slices approximately 2 mm thick along the coronal plane and placed in 0.25% TTC solution for staining in a 37°C incubator in the dark. During staining, the brain slices were occasionally turned with forceps to ensure uniform staining. After approximately 30 minutes of staining, the infarcted area appeared white, normal tissue appeared red, and the transition area between infarcted and normal tissue appeared pink. The brain slices were then fixed with 4% paraformaldehyde at 4°C for 24 h and photographed. The relative cerebral infarction rate for each mouse was calculated using IPP 6.0 software, as detailed in Formula I:

[0049] Formula I.

[0050] 5.2 Hematoxylin-eosin (HE) staining of mouse brain tissue: The paraffin blocks prepared in "Animal Sample Collection" were cut into 4 μm thin sections using a microtome, and then stained with HE and mounted with neutral resin. The stained sections were observed under a microscope, and the required images were collected and analyzed using an image acquisition system. Three samples were used in each group.

[0051] 5.3 Nissl staining of mouse brain tissue: The paraffin blocks prepared in the "Animal Sample Collection" section were cut into 4μm thin slices using a microtome. The slices were stained with Nissl staining solution and differentiated using 95% alcohol until clear Nissl bodies could be observed under a microscope. The slices were then mounted with neutral resin. The stained slices were observed under a microscope, and images were collected and analyzed using an image acquisition system. Three samples were used in each group.

[0052] 5.4 Detection of Apoptosis in Mouse Brain Tissue: The paraffin blocks prepared in the "Animal Sample Collection" section were sliced ​​into thin sections using a microtome. Apoptosis in mouse brain tissue was detected according to the TUNEL kit instructions. After DAPI staining, the sections were mounted in mounting media for preservation. Detailed observation and image recording were performed using a NIKON inverted fluorescence microscope. Three samples were included in each experimental group. This process aims to accurately identify and record apoptosis in brain tissue, providing an important foundation for subsequent data analysis.

[0053] 6. Experimental Results:

[0054] 6.1 Pharmacodynamic evaluation of Evodia Decoction in MCAO mice

[0055] According to the research plan ( Figure 1 Seven days after MCAO modeling (Chinese A), neurobehavioral scoring and statistical analysis were performed on mice in each group. Results are as follows: Figure 1In the control group (Figure B), no neurological function impairment was observed, and all mice scored 0. Compared with the control group, the MCAO group mice exhibited significant neurological deficits, such as circling and tipping, with significantly elevated neurological behavioral scores. Treatment with Wu Zhu Yu Tang significantly alleviated neurological function impairment, with scores decreasing in all treatment groups, indicating that Wu Zhu Yu Tang can improve neurological deficits caused by ischemic brain injury. TTC staining results of mouse brain tissue are shown below. Figure 1 As shown in C, Figure 1 In the diagram, the red area represents normal tissue, the white area represents the infarcted portion, and the pink area in the middle represents the ischemic transition zone. Results showed that no infarct foci were observed in the brain tissue of the control group mice, while the MCAO group mice showed significant infarct areas, with an average infarct area as high as 26.47%. After treatment with Evodia rutaecarpa decoction, the infarct area was significantly reduced, with the average infarct area in the 3 g / kg Evodia rutaecarpa decoction group decreasing to 4.75%, a decrease of 82.06% 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 the neurological function of MCAO mice and effectively reduce infarct volume, demonstrating its good neuroprotective effect against ischemic stroke and reperfusion injury. The 3 g / kg Evodia rutaecarpa decoction group showed the best efficacy and will be the key dose for future research on the anti-CIRI mechanism.

[0056] 6.2 Effects of Evodia Decoction on the 7-day survival status of MCAO mice

[0057] The protective effect of Wu Zhu Yu Tang on CIRI was further verified by assessing the survival rate and weight changes of MCAO mice. Figure 2 As shown in Figure A, the MCAO group had the lowest survival rate, only 40%, indicating that severe brain injury affects survival. Treatment with Evodia rutaecarpa decoction improved the survival rate, with the 3 g / kg decoction group showing the highest survival rate at 80%, demonstrating the best neuroprotective effect. The survival rate of the 6 g / kg group was 50%, lower than the 3 g / kg decoction group, suggesting that excessively high doses may have adverse effects. Mouse body weight changes are shown below. Figure 2 As shown in Figure B, the MCAO group mice experienced a significant decrease in body weight, possibly due to metabolic disorders and reduced food intake caused by ischemic injury. Treatment with Evodia rutaecarpa decoction promoted weight recovery, with the 3 g / kg decoction group showing the best recovery effect, indicating that this dose can effectively improve metabolic abnormalities caused by brain injury and improve quality of life. Evodia rutaecarpa decoction demonstrated a significant effect in improving the survival status of CIRI mice by increasing survival rate and promoting weight recovery. The 3 g / kg decoction group showed the best effect and is considered the optimal therapeutic dose. Subsequent experiments will further explore its anti-CIRI mechanism at this dose.

[0058] 6.3 Effects of Evodia Decoction on Pathological Morphological Changes in Brain Tissue of MCAO Mice

[0059] HE and Nissl staining were used to observe the effects of Evodia rutaecarpa decoction on the pathological morphological changes of brain tissue in MCAO mice. Results are as follows: Figure 3 As shown in Figure A, the left and right hemispheres of the control group mice maintained essentially the same color, with intact cell morphology, no edema or vacuoles, full cell nuclei, and clear nucleoli. In the model group mice, the infarct area of ​​the brain tissue was lighter in color than normal tissue, with severe edema, numerous scattered vacuoles, and shrunken and deeply stained nucleoli. Compared with the model group, administration of Wu Zhu Yu Tang significantly reduced brain tissue edema and nucleoli shrinkage in MCAO mice. Figure 3 As shown in Figure B, when observing the effects of Wu Zhu Yu Tang on Nissl bodies in the brain of MCAO mice, it was found that compared with the control group, the model group mice showed severe loss of Nissl bodies in the cerebral cortex, neuronal damage, and numerous vacuolar structures. Most neurons exhibited an atrophic phenotype, with weak staining and irregular distribution. After administration of Wu Zhu Yu Tang, the loss of Nissl bodies in the cortical area was significantly reduced, neuronal activity was greatly improved, no obvious vacuolar structures were found, and the neurons were darker in color and more orderly arranged. These findings indicate that Wu Zhu Yu Tang has a significant ameliorative effect on the pathological changes in the brain tissue of MCAO mice.

[0060] 6.4 Effects of Evodia Decoction on Neuronal Apoptosis in Brain Tissue of MCAO Mice

[0061] Neuronal apoptosis occurs during ischemic stroke, which is closely related to the severity of the disease. Therefore, the TUNEL assay was used to further detect the effect of Evodia rutaecarpa decoction on neuronal apoptosis in the brain tissue of MCAO mice. The results are as follows: Figure 4 As shown, no obvious TUNEL-positive cells were observed in the brain tissue of the control group mice, while the number of TUNEL-positive cells in the brain tissue of the MCAO group mice was significantly increased, indicating that MCAO modeling causes a large number of neuronal apoptosis; however, after administration of Wu Zhu Yu Tang, the number of TUNEL-positive cells in the brain tissue of mice was significantly reduced, showing that Wu Zhu Yu Tang has an anti-apoptotic effect on neuronal cells in MCAO mice.

[0062] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Use of a traditional 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 weight parts of traditional Chinese medicine materials: Evodia rutaecarpa 8-10 parts, Zingiber officinale 16-20 parts, Panax ginseng 8-10 parts, and Fructus jujubae 9-12 parts. The preparation method of the traditional Chinese medicine composition comprises extracting Evodia rutaecarpa, Zingiber officinale, Panax ginseng, and Fructus jujubae with water, and collecting the water extract; The temperature of the extraction is 95-100 DEG C; the extraction is performed for 2-4 times; the extraction time is 50-70 min / time; and after the extraction, the water extract is further subjected to vacuum concentration and drying to obtain a powder. The medicine has the pharmacological effects of improving the body weight of a patient and / or improving the survival index of a patient.

2. Use according to claim 1, characterized in that, The cerebral ischemia-reperfusion injury comprises at least one of the following injuries: nerve function injury, cerebral infarction, cerebral tissue edema, nerve cell apoptosis, reduced survival rate, and reduced body weight.

3. Use according to claim 1, characterized in that, The cerebral ischemia-reperfusion injury comprises middle cerebral artery occlusion induced by a thread method.

4. The use according to claim 1, characterized in that, The medicine also has the pharmacological effects of reducing the volume of cerebral infarction in cerebral ischemia-reperfusion injury, promoting the recovery of nerve function, and inhibiting nerve cell apoptosis.

5. The use according to claim 1, characterized in that, The traditional Chinese medicine composition is prepared from the following weight parts of traditional Chinese medicine materials: Evodia rutaecarpa 9 parts, Zingiber officinale 18 parts, Panax ginseng 9 parts, and Fructus jujubae 9 parts.

6. Use according to any one of claims 1 to 5, characterized in that, The dosage form of the medicine comprises the following kinds of oral preparations: tablets, powders, granules, capsules, decoctions, and oral liquids.

7. Use according to claim 6, characterized in that, The crude drug relative density of the traditional Chinese medicine composition in the medicine is 1.2-1.3 g / mL.

Images