CN111228323A

CN111228323A - Application of alcohol extract of pyrola in preparation of medicine for preventing and/or treating myocardial ischemic diseases

Info

Publication number: CN111228323A
Application number: CN202010208492.9A
Authority: CN
Inventors: 林蓉; 陷雨珊; 王建疆; 王维蓉; 杨广德; 郑子涵; 王冠; 姚凤; 靳真; 王博; 吕霄晗; 高鸿倩; 陈力方
Original assignee: Xinjiang Rongcheng Hack Pharmaceutical Co Ltd; Xian Jiaotong University
Current assignee: Xinjiang Rongcheng Hack Pharmaceutical Co Ltd; Xian Jiaotong University
Priority date: 2020-03-23
Filing date: 2020-03-23
Publication date: 2020-06-05

Abstract

The invention discloses application of a deer grass alcohol extract in preparing a medicament for preventing and/or treating myocardial ischemic diseases, belonging to the field of medicine and health. The alcohol extract of the deer grass can relieve oxidative stress and improve the energy metabolism level by activating Nrf2, thereby improving myocardial ischemia and hypoxia diseases; the alcohol extract of the deer grass is prepared by reflux extraction of ethanol water solution, and the prepared deer grass alcohol extract is a natural product with high curative effect and small side effect.

Description

Application of alcohol extract of pyrola in preparation of medicine for preventing and/or treating myocardial ischemic diseases

Technical Field

The invention belongs to the field of medicine and health, and relates to application of a deer grass alcohol extract in preparation of a medicine for preventing and/or treating myocardial ischemic diseases.

Background

Ischemic Heart Disease (IHD), also known as coronary atherosclerotic heart disease, is called coronary heart disease for short, and mainly refers to heart disease caused by myocardial ischemia and hypoxia due to coronary atherosclerosis, which causes obstruction of blood vessel lumen, imbalance between coronary blood supply and myocardial demand. In recent years, the prevalence rate of myocardial ischemia in China is on the rise year by year, the myocardial ischemia becomes a common disease and a frequently encountered disease of middle-aged and elderly people, and some young people aged 20-30 also have the myocardial ischemia. The pathogenesis of ischemic heart disease is complex and has not been fully elucidated so far. The theories of oxidative stress and energy metabolism are currently the more accepted important theories for explaining the pathogenesis of ischemic heart disease, and the ischemia and anoxia of myocardial tissues induce oxidative stress, generate a large amount of ROS and simultaneously interfere with the energy metabolism of myocardial cells, thereby causing myocardial damage.

At present, a plurality of medicines for clinically treating myocardial ischemia are available, such as: compound salvia dropping pills, coronary heart disease storax pills, quick-acting heart disease relieving pills and the like, but the traditional Chinese medicine compound has the defects of unclear mechanism, unstable curative effect, unobvious drug effect and the like, so that a safe and effective therapeutic medicine for myocardial ischemia is very necessary to be found.

Pyrola (Rhaponticus carthamoides (Willd.) IIjin, RC) is a plant of genus Rhaponticus of family Compositae, and is distributed in northwest, Siberian and middle Asia of Xinjiang, and is called Xinjiang ginseng. In folk, the deer grass is used as a tonic for treating diseases such as physical weakness, senile cardiac insufficiency, cardiovascular diseases and the like. In some research institutions such as russian medicinal plant and aromatic plant research institutes, the deer grass is used as a tonic, and meanwhile, the deer grass serving as a dietary supplement can be used for postoperative recovery, elimination of body weakness, increase of body energy level and the like. The latest research finds that the rhaponticum carthamoides has pharmacological effects of resisting cerebral ischemia, resisting arrhythmia and the like, and meanwhile, the rhaponticum carthamoides can increase the oxygen supply capacity.

The nuclear factor E2 related factor 2(nuclear factor erythroid 2p45-related factor 2, Nrf2) is the most active transcription regulator in the members of the CNC (cap 'n' coller) transcription factor family, can interact with an anti-oxidation response element ARE, and enhances the resistance of cells to oxidative stress. In recent years, research shows that Nrf2 not only is a main regulator of an antioxidant defense system of an organism and plays a role in oxidative stress, but also can be directly involved in processes such as regulation and control of key metabolic genes, or indirectly influence the expression of the genes through interaction with other transcription factors, and further participate in processes such as energy metabolism. In addition, researches prove that the traditional Chinese medicines and the traditional Chinese medicine compound such as the ligusticum wallichii, the qi-tonifying and blood-activating medicine can resist the oxidative stress injury of the myocardial ischemia rat by activating the Nrf2 channel.

Although the prior art reports that the deer grass has various biological activities, whether the deer grass can relieve myocardial ischemic diseases or not and relevant action mechanisms of the deer grass are not reported.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide the application of the deer grass alcohol extract in preparing the medicines for preventing and/or treating myocardial ischemic diseases.

In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:

the invention discloses application of a deer grass alcohol extract in preparing a medicament for preventing and/or treating myocardial ischemic diseases.

Preferably, the drug is a drug for treating an ischemic disease of the heart by improving the level of energy metabolism.

Further preferably, the drug is a drug that increases ATP and NAD in myocardial tissue⁺NADH content, and LD content in blood serum to improve energy metabolism level.

Preferably, the medicament is a medicament for treating cardiac ischemic diseases by improving oxidative stress level.

Further preferably, the drug is a drug that reduces oxidative stress by activating expression of Nrf2 protein in cardiac muscle cells.

More preferably, the drug is a drug for reducing the content of CK, CK-MB and AST in serum.

Further preferably, the deer grass alcohol extract is prepared by the following method:

1) pulverizing the root of herba Pyrolae, reflux-extracting with 70% ethanol water solution as solvent for several times (6 hr each time), and mixing extractive solutions to obtain ethanol extractive solution;

2) recovering solvent from the ethanol extractive solution, filtering, concentrating to obtain extract, oven drying, and pulverizing to obtain ethanol extract.

The invention also discloses a medicament for preventing and/or treating myocardial ischemic diseases, which is prepared from the alcohol extract of the deer grass and pharmaceutically acceptable pharmaceutic adjuvant into pharmaceutically acceptable dosage forms;

the deer grass alcohol extract is prepared by the following method:

Preferably, the pharmaceutical excipients comprise one or more of diluents, excipients, fillers, disintegrants, wetting agents, binders, surfactants, absorption enhancers and lubricants.

Preferably, the dosage form is a tablet, pill, capsule, granule or powder.

Compared with the prior art, the invention has the following beneficial effects:

the invention discloses an application of a deer grass alcohol extract in preparing a medicament for preventing and/or treating myocardial ischemic diseases, the deer grass alcohol extract can prolong the survival time of an anoxic mouse and relieve the blood pressure and ST segment change of an ischemic and anoxic rat, the deer grass alcohol extract has the function of reducing the content of CK, CK-MB and AST in rat serum and can reduce the damage of the rat myocardium and mitochondria induced by ISO, the deer grass alcohol extract has the function of reducing the content of LD in the rat serum, and the deer grass alcohol extract can treat the myocardial ischemic diseases by improving the energy metabolism level; the alcohol extract of the deer grass can reduce oxidative stress reaction by activating Nrf2, thereby improving myocardial ischemic diseases. The in vivo and in vitro pharmacological experiments prove that the wintergreen alcohol extract has the following pharmacological activities: (1) the effect of resisting myocardial ischemia and hypoxia of rats; (2) the effect of resisting the oxidative stress of rat cardiac muscle cells; (3) improving the energy metabolism of rat myocardial cells. The invention discovers that the rhaponticum carthamoides has good effect of treating myocardial ischemia and hypoxia, can be used for preventing and treating cardiovascular diseases, and has great clinical application value.

Drawings

FIG. 1 shows the effect of alcohol extract of Pyrola on normal pressure closed hypoxia/ISO-induced myocardial ischemia hypoxia mice, ISO-induced myocardial ischemia hypoxia rats ECG, blood pressure; wherein (a) is the influence of the alcohol extract of the deer grass on the survival time of mice with hypoxia under normal pressure; (b) influence of the alcohol extract of the deer grass on the survival time of the mice with myocardial ischemia and hypoxia induced by ISO; (c) influence of the alcohol extract of the deer grass on the change of the electrocardiogram S-T section of the mouse with myocardial ischemia and hypoxia induced by ISO; (d) the influence of the alcohol extract of the deer grass on the change of the cardiac ischemia hypoxia rat electrocardiogram S-T section induced by ISO is disclosed; (e) the influence of the alcohol extract of the deer grass on the heart rate change of the rats with myocardial ischemia and hypoxia induced by ISO is shown; (f) the influence of the alcohol extract of the deer grass on the change of the systolic pressure of the myocardial ischemia and hypoxia rats induced by ISO is disclosed; (g) the influence of the alcohol extract of the deer grass on the diastolic pressure change of the rats with myocardial ischemia and hypoxia induced by ISO; (h) is the influence of the alcohol extract of the deer grass on the average pressure change of the rats with myocardial ischemia and hypoxia induced by ISO.

FIG. 2 is a graph showing the effect of alcohol extract of Pyrola on the content of myocardial enzyme in rats with myocardial ischemia and hypoxia induced by ISO; wherein (a) is AST activity; (b) is CK activity; (c) is CK-MB activity.

FIG. 3 is a graph showing the results of HE staining of the deer grass alcohol extract on the myocardial tissue morphology of ISO-induced myocardial ischemia and hypoxia rats.

FIG. 4 is a transmission electron microscope photograph of mitochondrial morphology of alcohol extract of Pyrola japonica to ISO-induced rats with myocardial ischemia and hypoxia.

FIG. 5 is a graph showing the effect of alcohol extract of Pyrola on the energy metabolism index of myocardial tissue of rats with myocardial ischemia and hypoxia induced by ISO; wherein (a) is NAD⁺NADH; (b) is the LD content; (c) is the ATP content.

FIG. 6 is a graph showing the effect of alcohol extract of Pyrola on the oxidative stress index of myocardial tissue of rats with myocardial ischemia and hypoxia induced by ISO; wherein (a) is SOD content; (b) is the MDA content; (c) is CAT content; (d) is GSH-Px content.

FIG. 7 shows the alcohol extract of deer grass vs CoCl₂The effect of H9c2 injury from stimulation; wherein (a) the MTT method is used for detecting the influence of the deer grass alcohol extract on the activity of H9c 2; (b) is LDH activity; (c) is AST activity; (d) is CK activity; (e) is CK-MB activity.

FIG. 8 shows the alcohol extract of deer grass vs. CoCl₂Fluorescence microscopy of the effect of damaged rat cardiomyocytes.

FIG. 9 shows alcohol extract of deer grass vs CoCl₂A graph of the effect of oxidative stress indicators in damaged cardiomyocytes; wherein (a) is SOD content; (b) is the MDA content; (c) is CAT content; (d) is GSH-Px content.

FIG. 10 shows the alcohol extract of deer grass vs. CoCl₂An influence result graph of energy metabolism indexes in the damaged myocardial cells; wherein (a) is the ATP content; (b) is the LD content; (c) is NAD⁺/NADH。

FIG. 11 shows alcohol extract of deer grass vs CoCl₂Results of the effect of Nrf2 expression in injured rat cardiomyocytes.

FIG. 12 is Nrf2 vs CoCl₂Fluorescence microscopy of the effects of damaged cardiomyocytes.

FIG. 13 shows Nrf2 vs CoCl₂The effect of energy metabolism indicators in damaged cardiomyocytes; wherein (a) is the ATP content; (b) is the LD content.

Detailed Description

In order to make the technical solutions of the present invention better understood, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

It should be noted that the terms "first," "second," and the like in the description and claims of the present invention and in the drawings described above are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used is interchangeable under appropriate circumstances such that the embodiments of the invention described herein are capable of operation in sequences other than those illustrated or described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.

The invention is described in further detail below with reference to the accompanying drawings:

preparation of alcohol extract of herba Pyrolae

The extraction of the alcohol extract of the deer grass comprises the following steps:

1) taking 200g of dried rhaponticum carthamoides root, and crushing the rhaponticum carthamoides root into powder by using a crusher;

2) reflux-extracting with 70% ethanol water solution for three times, once for 6 hr, and mixing extractive solutions to obtain ethanol extractive solution;

3) and after recovering the ethanol, sequentially filtering, concentrating into thick paste, drying and crushing the thick paste to obtain the wintergreen ethanol extract.

Second, research on pharmacological action of alcohol extract of herba Pyrolae

Material sources are as follows: xinjiang Xinshengtai Biotechnology GmbH

1. The influence experiment of the alcohol extract of the deer grass on the normal pressure closed hypoxia/ISO induced myocardial hypoxia mice, ISO induced myocardial ischemia hypoxia rats ECG and blood pressure is as follows:

KM male mice irritate stomach and deer grass alcohol extract (60, 120, 240mg/kg) RC, positive medicine rhodiola rosea R.rosea (6mL/kg) for 14 days, put into 250mL wide-mouth bottles for closed hypoxia experiment, detect its survival time, as shown in (a) in figure 1, the result shows that deer grass alcohol extract and positive medicine rhodiola rosea can both prolong its survival time.

Modeling: KM male mice are injected with the alcohol extract of the deer grass (60, 120, 240mg/kg) and the rhodiola rosea (6mL/kg) as the positive drug for 14 days, and are injected with isoproterenol (20mg/kg) subcutaneously for 3 days to construct a mouse model with myocardial ischemia and hypoxia.

Detecting Electrocardiogram (ECG) changes before and after rat model building by using a BL-420S biological function experimental system; then the mixture is put into a 250mL wide-mouth bottle for a closed hypoxia experiment, and the survival time of the mixture is detected. As shown in (b) and (c) of FIG. 1, the results show that the alcohol extract of the deer grass and the rhodiola rosea which is a positive drug can improve the electrocardiogram change of mice and prolong the survival time of the mice.

Molding: SD male rats are injected with wintergreen alcohol extract (40, 80 and 160mg/kg) and rhodiola rosea (4mL/kg) as a positive drug for 14 days, and are injected with isoproterenol (85mg/kg) subcutaneously for 2 days to construct a specific myocardial ischemia and hypoxia rat model.

Detecting Electrocardiogram (ECG) changes before and after rat model creation by using BL-420S biological function experimental system, as shown in (d) in figure 1; detecting heart rate changes before and after rat model making by using a BP-300A full-automatic rat noninvasive blood pressure measuring system, as shown in (e) in figure 1; the BP-300A full-automatic rat noninvasive blood pressure measuring system is adopted to detect the blood pressure change before and after rat model making, as shown in (f), (g) and (h) in figure 1, the results show that the deer grass alcohol extract can improve the changes of rat electrocardiogram, heart rate and blood pressure.

2. The effect experiment of the alcohol extract of the deer grass on myocardial ischemia and hypoxia induced by ISO rat myocardial enzymes, myocardial tissue morphology and mitochondrial morphology comprises the following steps:

molding SD male rat according to the method, collecting blood from abdominal aorta 2 hr after molding, centrifuging at 3500rpm for 10min, collecting supernatant, and storing at-20 deg.C. The content of AST, CK and CK-MB is respectively detected by adopting Creatine Kinase (CK), creatine kinase isoenzyme (CK-MB) and glutamic-oxalacetic transaminase (AST) detection kits, the result is shown as (a), (b) and (c) in figure 2, and the result shows that compared with an ISO group, the pyrola alcohol extract and the rhodiola rosea which is a positive medicine can reduce the content of AST, CK and CK-MB in the serum of rats and reduce myocardial damage caused by ischemia and anoxia induced by ISO; killing rat, taking heart, flushing with normal saline to remove blood stain, removing non-myocardial tissue such as adipose tissue, cutting ventricle from apex to basal part of heart along atrioventricular groove, and trimming ventricular muscle tissue to about l mm³The small blocks are put into an electron microscope stationary liquid and used for detecting the mitochondrial morphology by a transmission electron microscope as shown in figure 4; the remaining ventricular muscle tissue was treated with 4% paraformaldehyde for HE staining to detect myocardial tissue morphology, as shown in fig. 3. The results show that the alcohol extract of the rhaponticum carthamoides and the rhodiola rosea serving as the positive medicine can improve the morphology of rat mitochondria and myocardial tissues.

3. The effect experiment of the alcohol extract of the deer grass on the myocardial tissue oxidative stress and energy metabolism of the rats with myocardial ischemia and hypoxia induced by ISO comprises the following steps:

about 100mg of myocardial tissue was taken, cold physiological saline was added, and a 10% myocardial homogenate was prepared using a tissue homogenizer. The homogenate was then centrifuged at 3000 rpm for 15 minutes at 4 ℃. Taking the homogenate supernatant, and detecting the contents of SOD, MDA, GSH-Px and CAT as oxidative stress indexes by using a determination kit of superoxide dismutase (SOD), Malondialdehyde (MDA), glutathione peroxidase (GSH-Px) and Catalase (CAT), as shown in figure 6, the result shows that the rhaponticum carthamoides alcohol extract can reduce the MDA content in myocardial tissue (figure 6 (b)), increase the contents of SOD in myocardial tissue (figure 6 (a)), CAT in figure 6 (c)) and GSH-Px in figure 6 (d)).

Detecting energy metabolism indexes ATP and NAD by using Adenosine Triphosphate (ATP), nicotinamide adenine dinucleotide (NAD (H)) and lactic acid (LD) determination kit⁺The results of the alcohol extract of wintergreen and rhodiola rosea, shown in FIG. 5, show that the LD content in rat serum can be reduced (FIG. 5 (b)), ATP in myocardial tissue (FIG. 5 (c)) and NAD (nicotinamide adenine dinucleotide)) can be increased⁺Content of NADH ((a) in FIG. 5).

4. Alcohol extract couple CoCl of rhaponticum carthamoides₂Effect of damaged cardiomyocytes experiment:

s1: collecting logarithmic phase cells, adjusting the concentration of cell suspension, adding 100 mu L of cell suspension into each hole of a 96-hole plate, and paving the plate to adjust the density of the cells to be detected to 1000-10000/hole;

S2：5％CO₂incubating at 37 deg.C until cell monolayer is 50% of the bottom of the well, adding alcohol extract of herba Pyrolae (100, 200, 400 μ g/mL) with different concentrations;

S3：5％CO₂incubation at 37 ℃ for 24h, CoCl was added₂(400μmol/L)；

S4: adding 10 μ L MTT solution (5mg/mL, i.e. 0.5% MTT) into each well, and continuing to culture for 4 h;

s5: the culture solution in the hole is sucked;

s6: adding 100 μ L of culture dimethyl sulfoxide into each well, shaking on a shaking table at low speed for 10min to dissolve the crystal completely, and measuring the light absorption value of each well at OD 490nm of an enzyme-labeling instrument.

As shown in FIG. 7, the results showed that the same CoCl was present₂Compared with the group, the wintergreen alcohol extract and the positive medicine salidroside improve the survival rate of hypoxic myocardial cells.

5. Alcohol extract couple CoCl of rhaponticum carthamoides₂Experiment of the effects of oxidative stress and energy metabolism in damaged cardiomyocytes:

h9c2 kinds of 6-pore plates, starving for 4H, pretreating with alcohol extract of herba Pyrolae (100, 200, 400 μ g/mL) for 24H, and adding CoCl₂After (400 mu mol/L) stimulation for 24h, collecting cells to detect the contents of SOD, GSH-Px, CAT and MDA in the cells, and processing the cells in the same wayAdding 1mL of diluted reagent DCFH-DA (serum-free culture solution dilution with dilution ratio of 1:1000) in Biyuntian ROS detection kit into 6-well plate of cells and 5% CO₂Incubate at 37 ℃ for 20 min. Cells were then washed 3 times with serum-free medium to remove DCFH-DA well without entering the cells. And finally, photographing through a fluorescence microscope. The results are shown in FIGS. 8 and 9.

The results show that compared with the control group, the hypoxia group significantly reduces the activities of SOD, CAT and GSH-Px in the myocardial cells and increases the MDA content and the ROS level, compared with the hypoxia group, the wintergreen alcohol extract group and the positive medicine salidroside group significantly increase the contents of SOD, CAT and GSH-Px in the myocardial cells, as shown in (a) (c) and (d) in figure 9, the ROS level and the MDA content are reduced, as shown in (b) in figures 8 and 9, and the wintergreen alcohol extract has the activity of resisting oxidative stress.

H9c2 kinds of 6-pore plates, starving for 4H, pretreating with alcohol extract of herba Pyrolae (100, 200, 400 μ g/mL) for 24H, and adding CoCl₂After 24h of (400. mu. mol/L) stimulation, cell supernatants were collected to detect LD content in the supernatants, and cells were collected to detect ATP, NAD/NADH content in the cells, as shown in FIG. 10.

The results show that compared with the control group, the hypoxia group obviously reduces the ATP and NAD/NADH levels and increases the LD content, and compared with the hypoxia group, the rhaponticum carthamoides alcohol extract administration group and the positive medicine salidroside group obviously increase the ATP and NAD⁺NADH level, and LD content is reduced, which indicates that the alcohol extract of the deer grass has the function of improving energy metabolism.

6. Alcohol extract couple CoCl of rhaponticum carthamoides₂Effect of Nrf2 expression in damaged cardiomyocytes experiment:

h9c2 kinds of 6-pore plates, starving for 4H, pretreating with alcohol extract of herba Pyrolae (100, 200, 400 μ g/mL) for 24H, and adding CoCl₂After stimulation for 24H (400. mu. mol/L), total protein of cells was extracted, and the expression level of Nrf2 in H9c2 was detected by Western blotting.

(1) The extraction steps of the rat myocardial cell protein are as follows:

s1: the medium was aspirated off, and the cells were gently rinsed 2 times with pre-cooled PBS;

s2: adding precooled RIPA lysate, adding protease inhibitor according to a ratio of 1:10, performing lysis on ice for 30min, and repeatedly blowing and beating cells, wherein each well is 200 mu L;

s3: collecting cell lysate, and centrifuging at 12000rpm for 20min at 4 ℃;

s4: the cell lysate supernatant was transferred to another new Eppendorf tube, and an appropriate amount was taken for protein concentration determination (protein quantification BCA kit), and the rest was stored at-80 ℃.

(2) Protein denaturation: adding 5 Xloading buffer solution into protein sample at ratio of 4:1, mixing, and decocting in boiling water for 5min to denature.

(3) Separation of the target protein: the target protein is separated by polyacrylamide gel electrophoresis (SDS-PAGE).

(4) Detection of the target protein: and detecting the target protein by adopting an ECL chemiluminescence method.

(5) Gel image analysis: and analyzing the gray value of the strip by adopting Lane 1D biological analysis software, and comparing the gray values of the target protein and the internal reference protein of the same sample to obtain a relative gray value.

As shown in fig. 11, the experimental results showed that the hypoxic group significantly reduced Nrf2 protein expression in cardiomyocytes compared to the control group; compared with the hypoxia group, the deer grass alcohol extract group and the salidroside group significantly increase the expression of Nrf2 protein in the cardiac muscle cells.

7. Nrf2 Pair CoCl₂Effects of oxidative stress and energy metabolism in injured cardiomyocytes experiment:

h9c2 kinds of 6-pore plates, starving for 4H, pretreating with alcohol extract of herba Pyrolae (100, 200, 400 μ g/mL) for 24H, and adding CoCl₂After stimulating by (400 mu mol/L) and adding an agonist SFN (2.5 mu mol/L) of Nrf2 for 24H, the expression levels of ROS, ATP and LD in H9c2 are detected by a kit.

The results are shown in fig. 12, and the experimental results show that the ROS level in the hypoxic cells is significantly increased compared to the control group; ROS levels in the cells of the rhaponticum carthamoides are significantly reduced compared to the hypoxic group, and the ROS levels in the cells are further reduced after activation of Nrf 2.

Results referring to fig. 13, experimental results show that activation of Nrf2 increased ATP content in hypoxic cardiomyocytes and decreased LD levels compared to the rhaponticum carthamoides group.

The invention takes a rat as a research object, establishes a rat model with myocardial ischemia and hypoxia by inducing and establishing isoproterenol (ISO, 85mg/kg/day) for 2 days through subcutaneous injection, and researches show that the rhaponticum carthamoides alcohol extract and the rhodiola rosea (Rhodiolarosa L, R. At the cellular level, rat cardiomyocytes H9c2 were studied by CoCl₂The method is characterized in that a myocardial cell hypoxia model is established through stimulation, the wintergreen alcohol extract and a positive medicine Salidroside (Salidroside, Sal) have the effect of resisting myocardial hypoxia, and further through exciting technologies such as Nrf2 and the like, the wintergreen alcohol extract is found to improve oxidative stress and energy metabolism through Nrf2, so that the effect of resisting myocardial hypoxia is achieved.

In conclusion, the invention takes the dry roots of the deer grass as the raw material and adopts 70 percent of ethanol water solution for reflux extraction to obtain the deer grass alcohol extract, and the invention has the following advantages: (1) the wintergreen alcohol extract is a natural product, and has high curative effect and small side effect; (2) the research of the invention finds that the rhaponticum carthamoides can treat myocardial ischemic diseases by improving the energy metabolism level; (3) the research of the invention finds that the rhaponticum carthamoides can relieve oxidative stress reaction by activating the Nrf2, thereby improving myocardial ischemic diseases.

The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.

Claims

1. Application of the alcohol extract of the deer grass in preparing the medicine for preventing and/or treating myocardial ischemia diseases.

2. The use according to claim 1, wherein the medicament is a medicament for treating an ischemic heart disease by improving the level of energy metabolism.

3. Use according to claim 2, characterized in that saidBy increasing ATP and NAD in myocardial tissue⁺NADH content, and LD content in blood serum to improve energy metabolism level.

4. The use of claim 1, wherein the medicament is a medicament for treating an ischemic cardiovascular disease by ameliorating a level of oxidative stress.

5. The use of claim 4, wherein the medicament is a medicament for alleviating oxidative stress by activating expression of Nrf2 protein in cardiomyocytes.

6. The use of claim 4, wherein said agent is an agent that reduces the levels of CK, CK-MB and AST in the serum.

7. The use of any one of claims 1 to 6, wherein the alcohol extract of deer grass is prepared by the following method:

8. A medicament for preventing and/or treating myocardial ischemic diseases is characterized in that the medicament is prepared into a pharmaceutically acceptable dosage form by a deer grass alcohol extract and pharmaceutically acceptable pharmaceutic adjuvants;

the deer grass alcohol extract is prepared by the following method:

9. The agent for preventing and/or treating myocardial ischemic diseases according to claim 8, wherein said pharmaceutical excipients comprise one or more of diluents, excipients, fillers, disintegrants, wetting agents, binders, surfactants, absorption enhancers and lubricants.

10. The agent for preventing and/or treating myocardial ischemic diseases according to claim 8, wherein said agent is in the form of a tablet, pill, capsule, granule or powder.