阅读说明：本技术 一种延丹胶囊在制备治疗心肌缺血药物中的应用 (Application of Yandan capsule in preparing medicine for treating myocardial ischemia ) 是由 何枢衡 于 2020-08-31 设计创作，主要内容包括：本发明公开了一种延丹胶囊在制备治疗心肌缺血药物中的应用,所述延丹胶囊可治疗由心肌缺血所引起的疾病包括心律失常、心肌梗死、冠心病,所述应用包括对延丹胶囊抗心肌缺血药效的验证方法,包括如下步骤：首先：对延丹胶囊对异丙肾上腺素诱导大鼠心肌缺血的作用及机制进行验证：本发明延丹胶囊主治气滞血瘀所致胸痹,可活血化瘀,理气宽胸,丹参作为活血化瘀的主药,对缺血性心脏病的预防和治疗具有显著的疗效。(The invention discloses an application of a delavay pill capsule in preparing a medicament for treating myocardial ischemia, wherein the delavay pill capsule can treat diseases caused by myocardial ischemia, such as arrhythmia, myocardial infarction and coronary heart disease, and the application comprises a method for verifying the anti-myocardial ischemia efficacy of the delavay pill capsule, and the method comprises the following steps: firstly: the effect and mechanism of the Yandan capsule on isoproterenol-induced myocardial ischemia of rats are verified: the Yandan capsule is mainly used for treating thoracic obstruction caused by qi stagnation and blood stasis, can promote blood circulation to remove blood stasis, regulates qi and relieves chest stuffiness, and the salvia miltiorrhiza is used as a main drug for promoting blood circulation to remove blood stasis, and has obvious curative effect on prevention and treatment of ischemic heart disease.)

1. Application of YANDAN Capsule in preparing medicine for treating myocardial ischemia is provided.

2. Use according to claim 1, characterized in that: the YANDAN Capsule can be used for treating diseases caused by myocardial ischemia, such as arrhythmia, myocardial infarction, and coronary heart disease.

3. Use according to claim 1, characterized in that: the application comprises a method for verifying the anti-myocardial ischemia efficacy of the Yandan capsule, which comprises the following steps:

firstly: the effect and mechanism of the Yandan capsule on isoproterenol-induced myocardial ischemia of rats are verified:

1. preparing materials;

1.1 drugs and reagents: yandan capsule; capsule for dredging heart meridian; isoproterenol hydrochloride; physiological saline injection; chloral hydrate; formaldehyde; MDA, T-SOD, AST detection kit; HIF-1. alpha. detection kit; IL-1 β (batch No. E20191001A), TNF- α detection kit; nrf2 antibody; HO-1 antibodies; a p53 antibody; bcl-2 antibodies; a Bax antibody; a GAPDH antibody;

1.2 animals: the method comprises the following steps of carrying out adaptive feeding on 60 SPF-grade SD rats with half male and female parts and 200 +/-20 g of body weight for 1 week at the temperature of 25 +/-2 ℃ and the relative humidity of 60 +/-5% under standard experimental conditions, then carrying out random grouping, and giving normal circadian rhythm to provide free drinking water and eating;

1.3 Instrument: BL-420N biological signal acquisition and analysis system; an analytical balance; a full-wavelength microplate reader; KD-BM biological tissue embedding machine; a Leica RM2235 microtome; an upright biological microscope BX 53; an EPS600 electrophoresis apparatus; a Western blot membrane transfer instrument; a full-automatic digital gel/chemiluminescence image analysis system;

2, the method comprises the following steps:

2.1 modeling and administration: randomly dividing 60 SD rats with half male and female into a blank group, a model group and a vein relaxing capsule group, wherein the dose is 200.57mg/kg, the low, medium and high dose groups of the Yandan capsule are 154, 308 and 616mg/kg, the dose in the Yandan capsule is clinical dose, the equivalent dose conversion ratio of human to rat is 1:6, the SD rat is continuously administered with gastric lavage for 8 days, the administration volume is 1ml/100g, the blank group and the model group are administered with equivalent physiological saline, on the 6 th to 8 th day, after 30min of gastric lavage administration, the rats in the other groups except the blank group are injected with ISO 40mg/kg in the abdominal cavity, and are continuously injected for 3d, and the blank group is injected with equivalent physiological saline for constructing an acute myocardial ischemia model;

2.2 electrocardiographic monitoring of rats with myocardial ischemia: after 30 mm of gastric lavage and administration on the 6 th day, 10 percent chloral hydrate anesthetizes each group of rats, after the rats enter an anesthetic state, the rats are injected with ISO, 40mg/kg in the abdominal cavity, and immediately perform limb II-lead electrocardiogram examination on the rats, and record the change of the ST segment;

2.3 serum factor assay: on the 8 th day, after the administration by gavage for 30min, incising through the abdominal midline of a rat, opening the abdominal cavity, collecting blood from the abdominal aorta into a sterile test tube, standing for 30min, centrifuging at 3000r/min for 15min, separating serum, determining the levels of MDA, T-SOD, AST, HIF-1 alpha, TNF-alpha and IL-1 beta, operating according to the method in each kit specification, and detecting by using a full-wavelength enzyme-labeling instrument;

2.4 myocardial histopathological examination: on the 8 th day, after the administration by gavage for 30min, the ventral midline of the rat is cut, the thoracic cavity is opened, the heart is taken down, the left ventricle tissue is selected, the left ventricle tissue is placed in a centrifugal tube filled with 10% formaldehyde solution for fixation for 48h, and the pathological change of the myocardial tissue is observed by adopting an HE staining method;

2.5Western Blotting: weighing 200mg of myocardial tissue, adding liquid nitrogen, fully grinding in a mortar, adding lysis buffer, fully mixing, cracking on ice for 30min, centrifuging at 12000 r/min for 30min at 4 ℃, and taking supernatant; carrying out protein quantification by using the BCA kit, and adjusting the protein concentration; after protein denaturation, loading, electrophoresis and membrane transfer; adding primary antibody for incubation overnight at 4 ℃, adding secondary antibody after TBST cleaning, incubating for 2h at room temperature, TBST cleaning, adding ECL luminescent liquid for exposure, scanning images, and analyzing experiment results by using Image J software, wherein the ratio of the gray value of the target protein to the gray value of the internal reference GAPDH is the relative content of the target protein;

2.6 immunohistochemistry: fixing 3 rat hearts in each group by using 10% formalin, dehydrating, embedding paraffin, conventionally slicing for 4 mu m, sequentially dewaxing, hydrating, repairing antigens, incubating primary antibodies, incubating secondary antibodies, staining hematoxylin nuclei, dehydrating by using gradient alcohol, clearing dimethylbenzene, sealing by using neutral gum, drying in the air, observing and photographing left ventricular cardiomyocytes under a microscope, and counting the positive cell rate of the pictures by using Image J software;

2.7 statistical analysis: except the electrocardiogram experiment result which adopts t test, the other data adopts one-factor variance analysis and is expressed by x +/-SEM, the statistical software is GraphPad Prism 5.01, and the P <0.05 is considered to have statistical difference;

3, analyzing results: by observing the change of an electrocardiogram ST segment of a rat with myocardial ischemia, the pathological change of myocardial tissues, the change of index levels such as MDA, AST, inflammatory factors and the like, the Yandan capsule can find out whether the Yandan capsule can effectively reduce the ST segment abnormal elevation of the rat with acute myocardial ischemia, reduce the pathological damage degree of the myocardial tissues, obviously reduce the MDA, AST, HIF-1 alpha and TNF-alpha levels in the serum of the rat and obviously increase the T-SOD level, and accordingly, whether the Yandan capsule has a good protection effect on the acute myocardial ischemia damage of the SD rat caused by isoproterenol is judged.

Technical Field

The invention relates to the technical field of traditional Chinese medicines, in particular to application of a delavay pill capsule in preparing a medicine for treating myocardial ischemia.

Background

In recent years, myocardial ischemic diseases seriously threaten human health, such as arrhythmia, myocardial infarction, coronary heart disease and the like. The incidence and the death rate of the medicine are on the rising trend year by year, and the life health and the safety of people are seriously influenced. The clinical symptoms of myocardial ischemia are usually manifested as palpitation, chest distress or chest pain, and the commonly used medicines comprise antiplatelet drugs, beta receptor blockers, calcium ion antagonists and the like. Because western medicines are easy to produce side effects after long-term administration, research and development of traditional Chinese medicines for treating myocardial ischemia are gradually becoming a trend.

According to clinical manifestations and pathogenesis, myocardial ischemia belongs to the categories of chest stuffiness, cardiodynia and the like in the traditional Chinese medicine theoretical system. The invention relates to a traditional Chinese medicine composition, which comprises the following raw medicinal materials: saviae Miltiorrhizae radix, rhizoma corydalis (processed with vinegar), Oletum Trogopterori, fructus Trichosanthis, Olibanum (processed with vinegar), radix Paeoniae alba, and fructus Aurantii. Mainly treats thoracic obstruction caused by qi stagnation and blood stasis, and can activate blood circulation to dissipate blood stasis, regulate qi and relieve chest stuffiness. The salvia miltiorrhiza, which is used as a main drug for promoting blood circulation and removing blood stasis, has a remarkable curative effect on prevention and treatment of ischemic heart disease, and has the treatment advantages of multiple target points and rich bioactive components. The main components of the composition are danshensu, salvianolic acid A, salvianolic acid B, tanshinone IIA and the like, which are important active ingredients for preventing and treating cardiovascular diseases. The total alkaloids and tetrahydropalmatine in rhizoma corydalis have effect in resisting arrhythmia and myocardial ischemia.

At present, most of treatment methods for myocardial ischemia are western medicines, interventional operations and the like, and have the effects of resisting platelet aggregation, stabilizing atherosclerotic plaques, improving blood supply of the heart and the like, but have the problems of drug resistance of the platelet medicines, toxic and side effects, poor perfusion of myocardial tissues, restenosis in a stent and the like. The Chinese medicinal preparation has definite curative effect on treating cardiovascular diseases, has low toxic or side effect and has good development prospect.

Disclosure of Invention

The invention aims to provide the application of the delavay pills in preparing the medicine for treating myocardial ischemia.

The invention is realized by the following technical scheme:

application of Yandan capsule in preparing medicine for treating myocardial ischemia

The YANDAN Capsule can be used for treating diseases caused by myocardial ischemia, such as arrhythmia, myocardial infarction, and coronary heart disease.

The application comprises a method for verifying the anti-myocardial ischemia efficacy of the Yandan capsule, which comprises the following steps:

firstly: the effect and mechanism of the Yandan capsule on isoproterenol-induced myocardial ischemia of rats are verified:

1. preparing materials;

1.1 drugs and reagents: yandan capsule; capsule for dredging heart meridian; isoproterenol hydrochloride; physiological saline injection; chloral hydrate; formaldehyde; MDA, T-SOD, AST detection kit; HIF-1. alpha. detection kit; IL-1 β (batch No. E20191001A), TNF- α detection kit; nrf2 antibody; HO-1 antibodies; a p53 antibody; bcl-2 antibodies; a Bax antibody; a GAPDH antibody;

1.2 animals: the method comprises the following steps of carrying out adaptive feeding on 60 SPF-grade SD rats with half male and female parts and 200 +/-20 g of body weight for 1 week at the temperature of 25 +/-2 ℃ and the relative humidity of 60 +/-5% under standard experimental conditions, then carrying out random grouping, and giving normal circadian rhythm to provide free drinking water and eating;

1.3 Instrument: BL-420N biological signal acquisition and analysis system; an analytical balance; a full-wavelength microplate reader; KD-BM biological tissue embedding machine; a Leica RM2235 microtome; an upright biological microscope BX 53; an EPS600 electrophoresis apparatus; a Western blot membrane transfer instrument; a full-automatic digital gel/chemiluminescence image analysis system;

2, the method comprises the following steps:

2.1 modeling and administration: randomly dividing 60 SD rats with half male and female into a blank group, a model group and a vein relaxing capsule group, wherein the dose is 200.57mg/kg, the low, medium and high dose groups of the Yandan capsule are 154, 308 and 616mg/kg, the dose in the Yandan capsule is clinical dose, the equivalent dose conversion ratio of human to rat is 1:6, the SD rat is continuously administered with gastric lavage for 8 days, the administration volume is 1ml/100g, the blank group and the model group are administered with equivalent physiological saline, on the 6 th to 8 th day, after 30min of gastric lavage administration, the rats in the other groups except the blank group are injected with ISO 40mg/kg in the abdominal cavity, and are continuously injected for 3d, and the blank group is injected with equivalent physiological saline for constructing an acute myocardial ischemia model;

2.2 electrocardiographic monitoring of rats with myocardial ischemia: after 30 mm of gastric lavage and administration on the 6 th day, 10 percent chloral hydrate anesthetizes each group of rats, after the rats enter an anesthetic state, the rats are injected with ISO, 40mg/kg in the abdominal cavity, and immediately perform limb II-lead electrocardiogram examination on the rats, and record the change of the ST segment;

2.3 serum factor assay: on the 8 th day, after the administration by gavage for 30min, incising through the abdominal midline of a rat, opening the abdominal cavity, collecting blood from the abdominal aorta into a sterile test tube, standing for 30min, centrifuging at 3000r/min for 15min, separating serum, determining the levels of MDA, T-SOD, AST, HIF-1 alpha, TNF-alpha and IL-1 beta, operating according to the method in each kit specification, and detecting by using a full-wavelength enzyme-labeling instrument;

2.4 myocardial histopathological examination: on the 8 th day, after the administration by gavage for 30min, the ventral midline of the rat is cut, the thoracic cavity is opened, the heart is taken down, the left ventricle tissue is selected, the left ventricle tissue is placed in a centrifugal tube filled with 10% formaldehyde solution for fixation for 48h, and the pathological change of the myocardial tissue is observed by adopting an HE staining method;

2.5Western Blotting: weighing 200mg of myocardial tissue, adding liquid nitrogen, fully grinding in a mortar, adding lysis buffer, fully mixing, cracking on ice for 30min, centrifuging at 12000 r/min for 30min at 4 ℃, and taking supernatant; carrying out protein quantification by using the BCA kit, and adjusting the protein concentration; after protein denaturation, loading, electrophoresis and membrane transfer; adding primary antibody for incubation overnight at 4 ℃, adding secondary antibody after TBST cleaning, incubating for 2h at room temperature, TBST cleaning, adding ECL luminescent liquid for exposure, scanning images, and analyzing experiment results by using Image J software, wherein the ratio of the gray value of the target protein to the gray value of the internal reference GAPDH is the relative content of the target protein;

2.6 immunohistochemistry: fixing 3 rat hearts in each group by using 10% formalin, dehydrating, embedding paraffin, conventionally slicing for 4 mu m, sequentially dewaxing, hydrating, repairing antigens, incubating primary antibodies, incubating secondary antibodies, staining hematoxylin nuclei, dehydrating by using gradient alcohol, clearing dimethylbenzene, sealing by using neutral gum, drying in the air, observing and photographing left ventricular cardiomyocytes under a microscope, and counting the positive cell rate of the pictures by using Image J software;

2.7 statistical analysis: except the electrocardiogram experiment result which adopts t test, the other data adopts one-factor variance analysis and is expressed by x +/-SEM, the statistical software is GraphPad Prism 5.01, and the P <0.05 is considered to have statistical difference;

3, analyzing results: by observing the change of an electrocardiogram ST segment of a rat with myocardial ischemia, the pathological change of myocardial tissues, the change of index levels such as MDA, AST, inflammatory factors and the like, the Yandan capsule can find out whether the Yandan capsule can effectively reduce the ST segment abnormal elevation of the rat with acute myocardial ischemia, reduce the pathological damage degree of the myocardial tissues, obviously reduce the MDA, AST, HIF-1 alpha and TNF-alpha levels in the serum of the rat and obviously increase the T-SOD level, and accordingly, whether the Yandan capsule has a good protection effect on the acute myocardial ischemia damage of the SD rat caused by isoproterenol is judged.

The invention has the advantages that:

according to clinical manifestations and pathogenesis, myocardial ischemia belongs to the categories of chest stuffiness, cardiodynia and the like in the traditional Chinese medicine theoretical system. The invention relates to a traditional Chinese medicine composition, which comprises the following raw medicinal materials: saviae Miltiorrhizae radix, rhizoma corydalis (processed with vinegar), Oletum Trogopterori, fructus Trichosanthis, Olibanum (processed with vinegar), radix Paeoniae alba, and fructus Aurantii. The traditional Chinese medicine composition is mainly used for treating thoracic obstruction caused by qi stagnation and blood stasis, can promote blood circulation to remove blood stasis, regulates qi and relieves chest stuffiness, and the salvia miltiorrhiza is used as a main medicine for promoting blood circulation to remove blood stasis, has obvious curative effect on prevention and treatment of ischemic heart disease, has the treatment advantages of multiple targets and rich bioactive components, and has the main components of tanshinol, salvianolic acid A, salvianolic acid B, tanshinone IIA and the like which are important active components for preventing and treating cardiovascular diseases, and the components of total alkaloids, tetrahydropalmatine and the like in corydalis tuber have the protection effects on arrhythmia and myocardial ischemia; in the prior art, most of treatment methods for myocardial ischemia are western medicines, interventional operations and the like, and have the effects of resisting platelet aggregation, stabilizing atherosclerotic plaques, improving blood supply of the heart and the like, but have the problems of drug resistance of the platelet medicines, toxic and side effects, poor perfusion of myocardial tissues, restenosis in a stent and the like. The Chinese medicinal preparation has definite curative effect on treating cardiovascular diseases, has low toxic or side effect and has good development prospect.

Drawings

FIG. 1 is a graph showing HE staining results (HE, x 40) of the heart of rats with myocardial ischemia in the control groups of the test by the Yandan capsule in the test of the present invention;

FIG. 2 is a graph showing the effect of Yandan capsule on the serum MDA, T-SOD and AST levels of myocardial ischemia rats in the control groups;

FIG. 3 is a graph showing the effect of Yandan capsule on the HIF-1 alpha level in serum of rats with myocardial ischemia in each control group in the test of the present invention;

FIG. 4 is a graph showing the effect of Yandan capsule on the levels of TNF-alpha and IL-1 beta in serum of myocardial ischemia rats of each control group in the test of the present invention:

FIG. 5 is a graph showing the effect of Yandan capsule on the Nrf2/HO-1/p53 signal channel-related protein in the heart tissue of myocardial ischemia rats of each control group in the invention test:

FIG. 6 is a graph showing the effect of Bcl-2/Bax protein expression level in heart tissue of rats with myocardial ischemia in each control group of Yandan capsules in the inventive test;

FIG. 7 is a graph of the effect of the Yandan capsule on the levels of Nrf2 and p53 protein in myocardial ischemia rat heart tissue in a test of the invention (x 40);

Detailed Description

As shown in figure 1, application of Yandan capsule in preparing medicine for treating myocardial ischemia

The YANDAN Capsule can be used for treating diseases caused by myocardial ischemia, such as arrhythmia, myocardial infarction, and coronary heart disease.

The application comprises a method for verifying the anti-myocardial ischemia efficacy of the Yandan capsule, which comprises the following steps:

firstly: the effect and mechanism of the Yandan capsule on isoproterenol-induced myocardial ischemia of rats are verified:

1. preparing materials;

1.1 drugs and reagents: yandan capsule; capsule for dredging heart meridian; isoproterenol hydrochloride; physiological saline injection; chloral hydrate; formaldehyde; MDA, T-SOD, AST detection kit; HIF-1. alpha. detection kit; IL-1 β (batch No. E20191001A), TNF- α detection kit; nrf2 antibody; HO-1 antibodies; a p53 antibody; bcl-2 antibodies; a Bax antibody; a GAPDH antibody;

1.2 animals: the method comprises the following steps of carrying out adaptive feeding on 60 SPF-grade SD rats with half male and female parts and 200 +/-20 g of body weight for 1 week at the temperature of 25 +/-2 ℃ and the relative humidity of 60 +/-5% under standard experimental conditions, then carrying out random grouping, and giving normal circadian rhythm to provide free drinking water and eating;

1.3 Instrument: BL-420N biological signal acquisition and analysis system; an analytical balance; a full-wavelength microplate reader; KD-BM biological tissue embedding machine; a Leica RM2235 microtome; an upright biological microscope BX 53; an EPS600 electrophoresis apparatus; a Western blot membrane transfer instrument; a full-automatic digital gel/chemiluminescence image analysis system;

2, the method comprises the following steps:

2.1 modeling and administration: randomly dividing 60 SD rats with half male and female into a blank group, a model group and a vein relaxing capsule group, wherein the dose is 200.57mg/kg, the low, medium and high dose groups of the Yandan capsule are 154, 308 and 616mg/kg, the dose in the Yandan capsule is clinical dose, the equivalent dose conversion ratio of human to rat is 1:6, the SD rat is continuously administered with gastric lavage for 8 days, the administration volume is 1ml/100g, the blank group and the model group are administered with equivalent physiological saline, on the 6 th to 8 th day, after 30min of gastric lavage administration, the rats in the other groups except the blank group are injected with ISO 40mg/kg in the abdominal cavity, and are continuously injected for 3d, and the blank group is injected with equivalent physiological saline for constructing an acute myocardial ischemia model;

2.2 electrocardiographic monitoring of rats with myocardial ischemia: after 30 mm of gastric lavage and administration on the 6 th day, 10 percent chloral hydrate anesthetizes each group of rats, after the rats enter an anesthetic state, the rats are injected with ISO, 40mg/kg in the abdominal cavity, and immediately perform limb II-lead electrocardiogram examination on the rats, and record the change of the ST segment;

2.3 serum factor assay: on the 8 th day, after the administration by gavage for 30min, incising through the abdominal midline of a rat, opening the abdominal cavity, collecting blood from the abdominal aorta into a sterile test tube, standing for 30min, centrifuging at 3000r/min for 15min, separating serum, determining the levels of MDA, T-SOD, AST, HIF-1 alpha, TNF-alpha and IL-1 beta, operating according to the method in each kit specification, and detecting by using a full-wavelength enzyme-labeling instrument;

2.4 myocardial histopathological examination: on the 8 th day, after the administration by gavage for 30min, the ventral midline of the rat is cut, the thoracic cavity is opened, the heart is taken down, the left ventricle tissue is selected, the left ventricle tissue is placed in a centrifugal tube filled with 10% formaldehyde solution for fixation for 48h, and the pathological change of the myocardial tissue is observed by adopting an HE staining method;

2.5Western Blotting: weighing 200mg of myocardial tissue, adding liquid nitrogen, fully grinding in a mortar, adding lysis buffer, fully mixing, cracking on ice for 30min, centrifuging at 12000 r/min for 30min at 4 ℃, and taking supernatant; carrying out protein quantification by using the BCA kit, and adjusting the protein concentration; after protein denaturation, loading, electrophoresis and membrane transfer; adding primary antibody for incubation overnight at 4 ℃, adding secondary antibody after TBST cleaning, incubating for 2h at room temperature, TBST cleaning, adding ECL luminescent liquid for exposure, scanning images, and analyzing experiment results by using Image J software, wherein the ratio of the gray value of the target protein to the gray value of the internal reference GAPDH is the relative content of the target protein;

2.6 immunohistochemistry: fixing 3 rat hearts in each group by using 10% formalin, dehydrating, embedding paraffin, conventionally slicing for 4 mu m, sequentially dewaxing, hydrating, repairing antigens, incubating primary antibodies, incubating secondary antibodies, staining hematoxylin nuclei, dehydrating by using gradient alcohol, clearing dimethylbenzene, sealing by using neutral gum, drying in the air, observing and photographing left ventricular cardiomyocytes under a microscope, and counting the positive cell rate of the pictures by using Image J software;

2.7 statistical analysis: except the electrocardiogram experiment result which adopts t test, the other data adopts one-factor variance analysis and is expressed by x +/-SEM, the statistical software is GraphPad Prism 5.01, and the P <0.05 is considered to have statistical difference;

3, results: electrocardiogram of myocardial ischemia damaged rat: after the intraperitoneal injection of ISO (40mg/kg), the electrocardiogram ST segment of the blank group of rats has no significant change. Compared with the blank group, the ST segment value of the rats in the model group is obviously increased (P < 0.01); compared with the model group, the electrocardiogram ST segment of the low, medium and high dose groups of the delayer capsule is remarkably reduced (P <0.05, P < 0.01);

see table 1.

TABLE 1 Effect of Yandan Capsule on myocardial ischemia rat electrocardiogram ST segment

Note: comparing with a blank control group by adopting a t test,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05，^**P<0.01。

3.2 the myocardial cells of the myocardial histopathology blank group are arranged orderly, the transverse striation is clear, the fibrous structure is not changed, and a large amount of inflammatory cells are not infiltrated; after Isoproterenol (ISO) is given to rats, the striations of a model group are broken, the arrangement of myocardial cells is disordered, and pathological changes such as myofilament lysis, a large amount of inflammatory cell infiltration and the like occur; the medium and high dose of the Yandan capsule can obviously improve the pathological changes of myocardial tissues;

3.2 myocardial histopathology: the blank group of myocardial cells are regularly and orderly arranged, the transverse striations are clear, the fiber structure is not changed, and a large amount of inflammatory cells are not infiltrated; after Isoproterenol (ISO) is given to rats, the striations of a model group are broken, the arrangement of myocardial cells is disordered, and pathological changes such as myofilament lysis, a large amount of inflammatory cell infiltration and the like occur; the middle and high dose of the Yandan capsule can obviously improve the pathological changes of myocardial tissues.

3.3 levels of MDA, T-SOD and AST in rat serum: compared with the blank group, the serum levels of MDA and AST of the model group rats are obviously increased (P <0.01), and the level of T-SOD is obviously reduced (P < 0.01). After different dose of delavay pills are given for pre-protection, the levels of MDA and AST in the serum of rats are obviously reduced (P is less than 0.01), and the level of T-SOD is obviously increased (P is less than 0.05, and P is less than 0.01).

3.4 HIF-1 alpha level in rat serum compared with blank group, HIF-1 alpha level in rat serum of model group is obviously increased (P < 0.01). And after different dose of Yandan capsules are given for pre-protection, the HIF-1 alpha level in the serum of rats is obviously reduced (P < 0.01).

3.5 the levels of TNF-alpha and IL-1 beta in the rat serum are obviously increased compared with the blank group (P is less than 0.01) in the rat serum of the model group. After different dosages of Yandan capsules are given for pre-protection, the TNF-alpha level in the serum of a rat is obviously reduced (P is less than 0.01); IL-1. beta. levels also tended to decrease, but were not statistically significant.

3.6 the effect of the Yandan capsules on the protein related to the Nrf2/HO-1/P53 signal channel is compared with that of a blank group, the expression levels of the model group Nrf2 and HO-1 protein are obviously reduced (P <0.01), and the expression level of the P53 protein is obviously increased (P < 0.05). After the minium capsule is taken, the expression levels of Nrf2 and HO-1 protein are obviously increased (P <0.05 and P <0.01), and the expression level of P53 protein is obviously reduced (P < 0.01).

3.7 Effect of Yandan capsules on Bcl-2/Bax protein levels the model group Bcl-2/Bax levels were significantly reduced (P <0.01) compared to the blank group. After the minium capsule is administrated, the Bcl-2/Bax level is obviously increased (P < 0.01).

3.8 immunohistochemistry results of the Nrf2 and P53 protein expression conditions of heart tissues show that the positive cell rate of the Nrf2 protein expression in heart tissues of a model group is obviously reduced compared with that of a blank group (P < 0.01). After the minium capsule is administrated, the protein expression rate is obviously increased (P < 0.05). The P53 showed a clear opposite trend, and the positive cell rate of P53 protein expression in heart tissue of the model group was significantly increased compared with that of the blank group (P < 0.01). After the minium capsule is taken, the positive cell rate of P53 protein expression is obviously reduced (P < 0.01).

4. And (4) conclusion:

by observing the change of the electrocardiogram ST segment of the myocardial ischemia rat, the pathological change of myocardial tissues, the change of index levels such as MDA, AST, inflammatory factors and the like, the Yandan capsule can effectively reduce the ST segment abnormal elevation of the acute myocardial ischemia rat, reduce the pathological damage degree of the myocardial tissues, obviously reduce the MDA, AST, HIF-1 alpha and TNF-alpha levels in the serum of the rat and obviously increase the T-SOD level. Therefore, the Yandan capsule has good protection effect on the acute myocardial ischemia injury of SD rats caused by Isoproterenol (ISO), and the effect of the Yandan capsule is related to the improvement of the cellular oxidative stress and the inflammation level.

As can be seen from Western blotting and IHC experimental results, the delaunay capsule can obviously reduce the expression of p53 and Bax proteins, and obviously improve the expression of Nrf2, HO-1 and Bcl-2 proteins. The Yandan capsule can improve the oxidative stress level and play a role in resisting apoptosis, thereby having an improvement effect on the acute myocardial ischemia injury of the rats caused by ISO.

In conclusion, the Yandan capsule has good protection effect on the acute myocardial ischemia injury of the rat caused by ISO, and the action mechanism of the Yandan capsule is probably related to factors such as oxidation resistance, inflammation resistance, cell apoptosis improvement and the like.