Medicine for inhibiting nerve cell apoptosis and improving ischemic stroke and application

Abstract

The invention discloses a medicine for inhibiting nerve cell apoptosis and improving ischemic stroke and application, and relates to the technical field of medical drugs, and the key point of the technical scheme is that Shenxiong pulse-invigorating granules inhibit nerve cell apoptosis and improve ischemic stroke by triggering a PI3K / Akt signal channel. The chemical components of the Shenxiong Tongmai granules are analyzed through ultra-high performance liquid chromatography / mass spectrometry (UPLC / MS), and the active components and the main action mechanism of the Shenxiong Tongmai granules for treating ischemic stroke are predicted in combination with network pharmacology and bioinformatics. And finally, the prediction result of bioinformatics is verified in the in-vitro cell model. Finally, it is found that the Shenxiong pulse-dredging granules can activate PI3K / AKT pathways in nerve cells, inhibit nerve cell apoptosis and improve ischemic stroke.

Description

Technical Field

[0001] The present invention relates to the technical field of medical drugs, and more specifically, to a drug for inhibiting nerve cell apoptosis and improving ischemic stroke and its application. Background Art

[0002] Ischemic stroke (IS) is the most common type of stroke in China, accounting for 60%-80% of the total incidence of stroke. Stroke is caused by a variety of reasons that result in cerebral blood supply disorders, leading to ischemia, hypoxia and necrosis of brain tissue, causing irreversible damage to brain tissue, and accompanied by corresponding neurological deficits. Stroke is characterized by high morbidity, high mortality and high disability rate, and is considered a major disease that threatens human life, health and safety. Stroke can be divided into hemorrhagic stroke (HS) and ischemic stroke (IS). At present, IS is the most common type of stroke in China, accounting for 60%-80% of the total incidence. Patients with acute IS are often treated with thrombolytic therapy in clinical practice. Currently, the only treatment approved by the US Food and Drug Administration (FDA) is tissue plasminogen activator (t-PA). However, t-PA has a narrow treatment window and there is a risk of further neurological damage. Therefore, it is necessary to find new alternative drugs with good efficacy and high safety.

[0003] For thousands of years, Chinese herbal medicines and their prescriptions have been used in China to treat a variety of diseases. More and more evidence confirms that Chinese herbal medicines are beneficial to human health and less toxic due to the co-evolution of organisms on Earth. Modern pharmacological studies have found that these natural medicines and their active ingredients can significantly improve the pathological changes in brain tissue of IS animals and reduce nerve damage. At the same time, the current clinical trial results show that SXTMG can significantly improve the SSS score and B1 score of IS patients, reduce the recurrence rate of cerebral infarction, the incidence of cerebral hemorrhage and the incidence of adverse reactions, indicating that SXTMG has obvious clinical efficacy and safety for IS. At the same time, animal model experiments also found that SXTMG can significantly reduce cerebral ischemia-reperfusion injury in rats and reduce the expression of P53 and Fas proteins as indicators of brain tissue apoptosis. However, the chemical composition and pharmacological mechanism of Shenxiong Tongmai Granules are still unclear, and the active ingredients and mechanisms of SXTMG against IS are still unclear, which seriously limits its clinical promotion and application.

[0004] Therefore, the present invention provides a drug and application for inhibiting neuronal apoptosis and improving ischemic stroke. Summary of the invention

[0005] The present invention establishes a MCAO animal model to evaluate the improvement effect of SXTMG on IS, and analyzes the main chemical components of SXTMG by ultra-high performance liquid chromatography / mass spectrometry (UPLC / MS). The main active ingredients and mechanisms of SXTMG's anti-IS effect are predicted by combining network pharmacology and bioinformatics analysis. Then an OGD / R cell model is established to verify the prediction results. Finally, it is found that compounds such as methyl salvianolic acid, salvianolic acid B, and salvianolic acid E may be the main chemical components of SXTMG in improving IS.

[0006] The present invention discloses a drug for inhibiting apoptosis of nerve cells and improving ischemic stroke and its application. The drug is Shenxiong Tongmai Granule, which is composed of Astragalus, Salvia miltiorrhiza and Chuanxiong. Shenxiong Tongmai Granule inhibits apoptosis of nerve cells and improves ischemic stroke by triggering PI3K / Akt signaling pathway, thereby improving brain damage caused by ischemic stroke.

[0007] In summary, the present invention has the following beneficial effects: The present invention analyzes the chemical components of Shenxiong Tongmai Granules by ultra-high performance liquid chromatography / mass spectrometry (UPLC / MS), and predicts the active components and main mechanisms of action of Shenxiong Tongmai Granules for the treatment of ischemic stroke by combining network pharmacology and bioinformatics. Finally, the prediction results of bioinformatics were verified in an in vitro cell model. Ultimately, it was found that Shenxiong Tongmai Granules can activate the PI3K / AKT pathway in neurons, inhibit neuronal apoptosis, and improve ischemic stroke. BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 It is that the Shenxiong Tongmai Granules in the embodiments of the present invention improve the ischemic symptoms of rats with middle cerebral artery occlusion (MCAO). Wherein, A and B show that the Shenxiong Tongmai Granules reduce the ischemic area of ​​the brain tissue of MCAO rats. C shows that the Shenxiong Tongmai Granules reduce the brain edema of MCAO rats. D shows that the Shenxiong Tongmai Granules reduce the neurological function damage score of MCAO rats. E and F show that the Shenxiong Tongmai Granules improve the pathological damage of the hippocampus of MCAO rats.

[0009] Figure 2 The Shenxiong Tongmai Granules in the embodiments of the present invention improve the pathological symptoms of rats with middle cerebral artery occlusion (MCAO). A shows that Shenxiong Tongmai Granules increase the number of Nissl bodies in the hippocampus of MCAO rats; B shows that Shenxiong Tongmai Granules reduce the level of oxidative stress in the brain tissue of MCAO rats.

[0010] Figure 3 The UPLC / MS test results of Shenxiong Tongmai Granules in the embodiment of the present invention are shown in Figure 1. A is the ESI+ mode detection spectrum of Shenxiong Tongmai Granules; B is the ESI- mode detection spectrum of Shenxiong Tongmai Granules; and C is the structural formulas of 22 compounds identified in Shenxiong Tongmai Granules.

[0011] Figure 4 This is the network pharmacology analysis of Shenxiong Tongmai Granules in the embodiment of the present invention. A shows the intersection of the ischemic stroke (IS) target library and the Shenxiong Tongmai Granules target library; B is the IS-target-ingredient-Shenxiong Tongmai Granules network diagram; C is the interaction network diagram of the intersection genes.

[0012] Figure 5 It is the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the target of Shenxiong Tongmai Granule in the embodiment of the present invention. Among them, A and B are the GO analysis of the intersection genes; C and D are the KEGG analysis of the intersection genes.

[0013] Figure 6 It is that Shenxiong Tongmai Granules in the embodiment of the present invention protect HT22 cells from oxygen glucose deprivation / reoxygenation (OGD / R) damage. Among them, A shows that Shenxiong Tongmai Granules intervention does not affect the cell viability of HT22 cells; B shows the establishment of OGD / R cell model and the flow chart of Shenxiong Tongmai Granules intervention; C shows the changes in cell morphology and cell viability after Shenxiong Tongmai Granules intervention.

[0014] Figure 7 It is that the Shenxiong Tongmai granules in the examples of the present invention reduce the expression of proteins related to apoptosis in HT22 cells.

[0015] Figure 8 It is that the Shenxiong Tongmai granules in the examples of the present invention upregulate the phosphorylation level of phosphatidylinositol 3-kinase (PI3K) in HT22 cells.

[0016] Fig. 9 It is that the Shenxiong Tongmai granules in the examples of the present invention upregulate the phosphorylation level of protein kinase B (AKT) in HT22 cells. DETAILED DESCRIPTION

[0017] The following is combined with Figure 1-9 The present invention is described in further detail.

[0018] Embodiment: A drug for inhibiting neuronal apoptosis and improving ischemic stroke and its application, such as Figure 1-Figure 9 shown.

[0019] Ischemic stroke (IS) is the most common type of stroke in China, accounting for 60%-80% of the total incidence of stroke. Shenxiong Tongmai Granule (SXTMG) is one of the commonly used prescriptions in traditional Chinese medicine. This example studies the possible active substances and molecular mechanisms of SXTMG against IS. First, the anti-IS effect of SXTMG was demonstrated in a rat model of middle cerebral artery occlusion (MCAO), and the composition of SXTMG was determined based on high performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS). Bioinformatics analysis of the identified components was performed in combination with the online database of disease targets to predict potential molecular mechanisms, and the results showed that apoptosis and PI3K / Akt were potential pathways for SXTMG to resist IS. Subsequently, the predicted results of bioinformatics analysis were confirmed using an in vitro glucose-oxygen deprivation / reoxygenation (OGD / R) cell model, and the results showed that SXTMG could inhibit OGD / R-induced apoptosis of HT22 cells. The results confirmed that SXTMG treatment can activate the PI3K / Akt signaling pathway in OGD / R-induced HT22 cells. Experiments have shown that Shenxiong Tongmai Granules (SXTMG) inhibits neuronal apoptosis by triggering the PI3K / Akt signaling pathway and has an improving effect on ischemic stroke.

[0020] Materials and Methods

[0021] Materials: The main ingredients of Shenxiong Tongmai Granules are 15g-45g of Astragalus, 10g-20g of Salvia and 5g-15g of Chuanxiong, specifically 30g of Astragalus, 15g of Salvia and 10g of Chuanxiong. Antibodies to p-Akt, Akt, p-PI3K, PI3K, Bax, Bcl-2, c-Caspase3 and c-Caspase9 were purchased from Abcam.

[0022] Preparation of SXTMG freeze-dried powder: SXTMG decoction-free granules were provided by the pharmacy of the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine. Add 10 times the amount of water to the granules, heat and stir, fully dissolve, and filter while hot. Freeze-dry the filtrate to obtain SXTMG freeze-dried powder.

[0023] Animal model and treatment: 43 male SD rats (220±5g) were purchased from Chengdu Dashuo Experimental Animal Center. The animals were kept in a stable environment with a temperature of 22℃~25℃, a humidity of 45%~65%, and a 12-hour day and night alternation. The animals were adaptively fed for 7 days before the start of the experiment, and the animal study was completed under the supervision and approval of the Animal Protection and Ethics Committee of Chengdu University of Traditional Chinese Medicine (No.: 2020-07-4). Rats were randomly divided into three groups, including a control group, an MCAO group, and a SXTMG treatment group. Before MCAO modeling, the rats were pretreated. Rats in the SXTMG group were given 1.2mg / kg SXTMG freeze-dried powder, and rats in the sham operation group and MCAO group were given an equal amount of normal saline for 14 days. On the 15th day, the MCAO rat model was prepared according to the method established in the study.

[0024] Neurological function score: 24 hours after perfusion, the neurological function of the rats was evaluated according to the following criteria: 0 point, no nerve damage; 1 point, the contralateral forelimb could not be extended; 2 points, the contralateral forelimb was flexed; 3 points, slight head turn to the contralateral side; 4 points, no spontaneous activity and impaired consciousness.

[0025] Brain water content: At the end of the experiment, the rats were killed, the brains were collected, and the wet weight of the brain tissue was recorded. The brain tissue was then dried in an oven at 105°C for 24 hours, and the dry weight of the brain tissue was recorded. Finally, the brain tissue water content was calculated according to the following formula: Water content = (wet weight - dry weight) / wet weight × 100 (%).

[0026] TTC staining: After the experiment, collect the animal brain tissue and freeze it in a -20℃ refrigerator for about 20 minutes. Cut the frozen brain tissue into 2mm thick slices. Then, put the slices into 2% TTC solution, cover them with tin foil to avoid light, and put them in a 37℃ incubator for 15-30min. Turn the brain slices every 5min to make them evenly contact the staining solution. After staining, take out the brain slices and wash them with PBS solution for 3-5min. Finally, collect images of the stained brain slices and calculate the infarct area.

[0027] H&E staining: At the end of the experiment, the animal brain tissues were collected, fixed with 4% paraformaldehyde (PFA), and then embedded in paraffin and sectioned. The pathological changes of brain tissues were observed by H&E staining according to the reported method.

[0028] Nissl staining: At the end of the experiment, the brain tissues of the animals were collected, fixed with 4% paraformaldehyde (PFA), and then embedded in paraffin and sectioned. The changes in the number of Nissl bodies in the brain tissues were observed by Nissl staining.

[0029] Detection of oxidative stress indicators: After the animal experiment, the animal brain tissue was collected and quickly frozen in liquid nitrogen. At the beginning of the experiment, the brain tissue was taken out from the -80°C refrigerator, ground in tissue lysis buffer, and then centrifuged to obtain the supernatant. The levels of catalase (CAT), glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) in brain tissue were detected using commercial kits according to the instructions.

[0030] UPLC / MS analysis: Take 2 mg of freeze-dried powder, add 5 mL of methanol to reconstitute, take 1 mL of the solution and filter it with a microporous filter membrane (0.22 μm) as a sample for mass spectrometry detection. The sample components were analyzed by ultra-high performance liquid chromatography-quadrupole-orbitrap mass spectrometry (UPLC-QE-Orbitrap-MS, Thermo, USA). The chromatographic column was a Thermo Fisher Scientific Accucore C18 column (100 mm x 2.1 mm, 2.6 μm). The column temperature was set to 35 ° C, the mobile phase flow rate was 1.0 mL / min, and the injection volume was 10 μL. The mobile phase was composed of acetonitrile (A) and 0.01% formic acid solution (B), and gradient elution was used, as follows: 0-6 min, 5%-20% A; 6-11 min, 20%-22% A; 11-20 min, 22%-60% A; 20-25 min, 60%-65% A; 25-28 min, 65%-65% A; 28-30 min, 65%-95% A. Electrospray ionization source (ESI) and positive and negative ion mode scanning were used for mass spectrometry detection, with a scanning range of 100-150 m / z. The model of the sampler was HPLC-MS / MS, with a scanning range of 100-1500 m / z. The capillary mass spectrometer temperature was set at 320°C, nitrogen was used as the desolvation gas, and the detection flow rate was 1.0 L / h.

[0031] Bioinformatics analysis: The targets of the compounds identified in the mass spectrometry experiment were predicted using an online drug target prediction database as the target library of SXTMG. The IS-related targets used were obtained from the GeneCards and OMIM databases as the target library of IS. First, the overlapping targets of the SXTMG target library and the IS target library were obtained. These targets were considered to be possible targets for SXTMG to improve IS, and the SXTMG-compound-IS network was established. The interaction network of these overlapping targets was studied using String online software. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed on the potential pathways of the overlapping targets using the DAVID online analysis tool.

[0032] Cell culture and intervention: HT22 cells were purchased from Guangzhou Genio Biotechnology Co., Ltd. and cultured in DMEM medium supplemented with 10% fetal bovine serum. To determine the optimal working concentration of SXTMG, the CCK8 kit was first used to measure the cell viability after 24 hours of SXTMG intervention. A glucose-oxygen deprivation / reoxygenation (OGD / R) model of cells was established. HT22 cells were first placed in an anaerobic chamber and sugar-free medium for 4 hours. The cells were then transferred to normoxic conditions and treated with normal DMEM medium, while different doses of SXTMG were added for 12 hours. At the end of the treatment, the survival rate of each group of cells was measured using the CCK8 kit.

[0033] WB detection: After treatment, cells were collected and cell lysis buffer was added to extract total cell protein. WB experiments were performed according to the method described by Peng et al. to detect the expression of apoptosis-related proteins in cells.

[0034] Immunofluorescence staining: Cells in the logarithmic growth phase were inoculated into confocal laser culture dishes. After the cells were completely attached, the cells were treated with OGD / R and SXTMG. At the end of the treatment, the cells were fixed with 4% paraformaldehyde (PFA) and then blocked with 5% bovine serum albumin (BSA). After blocking, the corresponding primary antibody was added and incubated overnight, and then the corresponding fluorescent secondary antibody was added and incubated at room temperature for 1 hour. After incubation, the cell nucleus was fluorescently labeled with DAPI. Finally, the expression of proteins in the cells was observed using a confocal laser microscope.

[0035] Statistical analysis: Statistical analysis was performed using SPSS 20.0 and GraphPad 9.0, and the results were expressed as mean ± standard deviation. One-way or two-way analysis of variance (ANOVA) followed by Tukey's multiple comparison test was used to compare multiple groups. P values ​​less than 0.05 were considered statistically significant.

[0036] result

[0037] SXTMG improves reperfusion brain injury in MCAO rats

[0038] The MCAO rat model can well simulate cerebral ischemia-reperfusion injury. In this example, a MCAO rat model was constructed to study the protective effect of SXTMG on cerebral ischemia-reperfusion injury. Figure 1 As shown in Figure 2, compared with the sham operation group, the rat brain tissue after MCAO surgery showed obvious ischemic areas ( Figure 1 A). However, the rats that were pre-treated with SXTMG had a significantly reduced ischemic area after MCAO surgery, indicating that SXTMG has a significant protective effect against cerebral ischemia-reperfusion injury in rats ( Figure 1 A&B). At the same time, the hydrocephalus of the rats was detected, and the results showed that MCAO surgery significantly increased the volume of hydrocephalus in the rats, but pre-treatment with SXTMG significantly reduced the increase in hydrocephalus caused by MCAO surgery ( Figure 1 C) At the end of the experiment, the rats were also evaluated for neurological impairment. Figure 1 As shown in Figure D, MCAO surgery can cause neurological damage in rats. SXTMG treatment significantly reduced the neurological damage in rats. In addition, H&E staining was performed on the rat brain tissue to study the pathological changes in the rat brain tissue. Figure 1As shown in E&F, MCAO surgery caused shrinkage, reduced size, dark nuclear staining and disordered arrangement of hippocampal pyramidal cells in the CA1, CA3 and DG regions of the rat hippocampus. Pre-treatment with SXTMG significantly improved these pathological changes. Similarly, Nissl staining results also showed that MCAO significantly reduced the number of Nissl bodies in hippocampal pyramidal cells, while SXTMG increased the number of Nissl bodies in the CA1, CA3 and DG regions of rats ( Figure 2 A). Excessive oxidative stress in brain tissue is one of the main causes of cerebral ischemia-reperfusion injury. Therefore, the effect of SXTMG on the level of oxidative stress in brain tissue of MCAO rats was also studied. Figure 2 As shown in B, MCAO surgery significantly reduced the levels of antioxidant enzymes such as SOD, CAT, and GSH in rat brain tissue and significantly increased the level of oxidative stress product MDA. However, the levels of antioxidant enzymes in the brain tissue of rats treated with SXTMG in advance were significantly increased, and the level of MDA was significantly reduced. In conclusion, pre-treatment with SXTMG can significantly reduce brain tissue damage caused by ischemia-reperfusion and reduce the level of oxidative stress in brain tissue.

[0039] Analysis of Chemical Components of Shenxiong Tongmai Granules

[0040] The chemical components in Shenxiong Tongmai Granules (SXTMG) were detected by ultra-performance liquid chromatography / mass spectrometry (UPLC / MS). Figure 3 As shown. Within 50 minutes, more than 40 compounds were detected and 22 compounds were identified, including danshensu, caffeic acid, ferulic acid, salvianolic acid, salvianolic acid D, salvianolic acid methyl ester, rosmarinic acid, salvianolic acid B, salvianolic acid E, salvianolic acid A, calycosin, salvianolic acid C, astragaloside III, formononetin, astragaloside IV, astragaloside II, ligustilide A, danshenxinkun A, astragaloside I, tanshinone II B, tanshinone I, and tanshinone II A.

[0041] Bioinformatics analysis of Shenxiong Tongmai granules in improving brain damage in rats with middle cerebral artery occlusion

[0042] More than 3,000 SXTMG-related targets and 751 ischemic stroke (IS)-related targets were identified through database searches. The target library of SXTMG and the target library of ischemic stroke intersected to obtain 89 common targets, which are considered to be potential targets for SXTMG to improve ischemic stroke ( Figure 4 A). In addition, a target network of Shenxiong Tongmai Granules-ischemic stroke was constructed, and it was found that the most targets interacted with compounds such as salvianolic acid methyl ester, salvianolic acid B, and salvianolic acid E. This indicates that these compounds are the main chemical components of Shenxiong Tongmai Granules that improve ischemic stroke ( Figure 4B), methyl salvianolic acid has an improving effect on ischemic stroke. At the same time, the interactions of these 89 common targets were analyzed, and it was found that AKT1, SRC, MAPK1, TNF, CASP3 and other targets had the most interactions, which indicates that these targets may be the most important targets for Shenxiong Tongmai Granule to improve ischemic stroke ( Figure 4 C). Finally, pathway analysis of common targets was performed and it was found that most of these targets were concentrated in the PI3K-AKT pathway and apoptosis pathway, suggesting that Shenxiong Tongmai Granules may play a role in improving ischemic stroke by acting on these pathways. Shenxiong Tongmai Granules protects against neurological damage in rats with middle cerebral artery occlusion by activating the PI3K / AKT pathway ( Figure 5 )

[0043] Shenxiong Tongmai Granule protects against neurological injury in rats with middle cerebral artery occlusion by activating PI3K / AKT pathway

[0044] To verify whether the neuroprotective effect of Shenxiong Tongmai Granules on ischemic stroke is related to the PI3K-AKT pathway, an in vitro oxygen-glucose deprivation / reoxygenation (OGD / R) cell model was established. First, the cytotoxicity of Shenxiong Tongmai Granules at different concentrations on HT22 cells was detected. The results showed that 0-320 μg / mL Shenxiong Tongmai Granules did not affect the cell viability of HT22 cells (≥90%) ( Figure 6 A). In subsequent experiments, 40, 80, and 160 μg / mL were selected as the therapeutic concentrations of Shenxiong Tongmai Granules. In subsequent experiments, it was found that glucose-oxygen deprivation / reoxygenation treatment significantly reduced the cell viability of HT22 cells ( Figure 6 B), but Shenxiong Tongmai Granules restored the cell viability of HT22 cells, indicating that Shenxiong Tongmai Granules had a protective effect on HT22 cells ( Figure 6 C). Next, we studied whether the cytoprotective effect of Shenxiong Tongmai Granules was related to the inhibition of cell apoptosis. Western blot (WB) experiments showed that glucose-oxygen deprivation / reoxygenation intervention significantly increased the expression of apoptosis-related proteins BAX, c-Caspase3, and c-Caspase9, and reduced the expression of anti-apoptotic protein Bcl-2 ( Figure 7 ). Fortunately, treatment with Shenxiong Tongmai Granules reversed these conditions, indicating that the cytoprotective effect of Shenxiong Tongmai Granules is related to anti-apoptosis. Network pharmacology analysis showed that the neuroprotective effect of Shenxiong Tongmai Granules on ischemic stroke is related to the regulation of the PI3K / Akt pathway. Therefore, in the in vitro experiment, the expression of PI3K / AKT pathway-related proteins in HT22 cells was detected, and the results were as follows Figure 8 and Fig. 9As shown. Glucose-oxygen deprivation / reoxygenation treatment significantly inhibited the phosphorylation of PI3K and Akt in HT22 cells, indicating that the PI3K / AKT pathway in the cells was inhibited. Inhibition of the PI3K / AKT pathway can lead to cell apoptosis, and inhibition of the PI3K / AKT pathway is also the main cause of apoptosis in ischemic stroke-type neurons. After intervention with Shenxiong Tongmai Granules (SXTMG), the levels of p-PI3K and p-AKT in the cells were significantly increased, indicating that SXTMG can activate the PI3K / AKT pathway. In conclusion, SXTMG can inhibit neuronal apoptosis, and its mechanism is related to the activation of the PI3K / AKT pathway, which is consistent with the results predicted by previous network pharmacology.

[0045] In this example, the chemical components of Shenxiong Tongmai Granules were analyzed by ultra-high performance liquid chromatography / mass spectrometry (UPLC / MS), and the active ingredients and main mechanisms of action of Shenxiong Tongmai Granules for the treatment of ischemic stroke were predicted by combining network pharmacology and bioinformatics. Finally, the prediction results of bioinformatics were verified in an in vitro cell model. Ultimately, it was found that Shenxiong Tongmai Granules can activate the PI3K / AKT pathway in neurons, inhibit neuronal apoptosis, and improve ischemic stroke.

[0046] This specific embodiment is merely an explanation of the present invention and is not a limitation of the present invention. After reading this specification, those skilled in the art may make non-creative modifications to the present embodiment as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by the patent law.

Claims

1. A drug for inhibiting neuronal apoptosis and improving ischemic stroke and its application, characterized in that: The medicine is Shenxiong Tongmai Granules, which are composed of 30 grams of Astragalus, 15 grams of Salvia miltiorrhiza and 10 grams of Chuanxiong.

2. Use of the Shenxiong Tongmai Granule according to claim 1 in the preparation of a drug for improving ischemic stroke by inhibiting neuronal apoptosis by triggering the PI3K / Akt signaling pathway.

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