Application of aveheart in prevention or treatment of mitochondrial abnormality diseases

Abstract

The invention discloses an application of Evaixin in prevention or treatment of mitochondrial abnormality diseases. The invention provides an application of an Avaixin oral liquid in preparation of a product for improving mitochondrial homeostasis imbalance. The mitochondrial homeostasis imbalance is caused by cardiovascular diseases. The invention proves that the Avaifuxin oral liquid has a potential protection effect on cardiovascular diseases, and plays a protection role by adjusting the dynamic balance of mitochondrial fusion and division processes. The invention is of great significance to further understanding the protective action mechanism of the Aiwei heart oral liquid on the mitochondrial homeostasis imbalance caused by cardiovascular diseases (hypertensive myocardial hypertrophy, myocardial fibrosis, ventricular remodeling, heart failure, myocardial infarction and myocardial ischemia-reperfusion injury) and improving the corresponding curative effect.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the use of Avicel for preventing or treating mitochondrial abnormality diseases. Background Art

[0002] Cardiovascular diseases include hypertension, myocardial hypertrophy, myocardial fibrosis, ventricular remodeling, heart failure, myocardial infarction, myocardial ischemia-reperfusion injury, etc.

[0003] Myocardial ischemia-reperfusion injury (MIRI) is a phenomenon in heart disease in which restoration of blood flow (reperfusion) to a part of the heart after ischemia due to blood flow obstruction may exacerbate cardiac damage rather than simply restore tissue function. This phenomenon is very common in clinical treatments such as myocardial infarction, such as restoration of coronary blood flow through thrombolytic therapy or percutaneous coronary intervention. The mechanism of MIRI is complex and involves multiple molecular and cellular processes. During ischemia, cardiomyocytes are damaged by lack of oxygen and insufficient nutrient supply. After blood flow is restored, it is expected to relieve the hypoxia, but in fact, reperfusion induces a series of reactions, including the massive production of reactive oxygen species, intracellular calcium overload, mitochondrial dysfunction, and inflammatory response, which together lead to further damage to cardiomyocytes. The severity of MIRI is affected by multiple factors, including the duration of ischemia, the speed and pattern of blood flow restoration, and the accompanying risk factors. For example, prolonged ischemia can lead to more extensive cardiomyocyte death and tissue damage, whereas rapid and complete reperfusion may induce a more dramatic burst of reactive oxygen species and calcium overload than slow or incomplete reperfusion.

[0004] As the energy factory of the cell, the homeostasis of mitochondria's function and structure is essential for cell survival. Mitochondrial homeostasis involves dynamic processes such as mitochondrial biogenesis, fission, fusion, autophagy, and clearance, which together maintain the morphology, function, and quality control of the mitochondrial network. Mitochondrial fusion and fission are two key processes that regulate mitochondrial morphology and function, affecting the stability of mitochondrial DNA and cellular energy metabolism. Mitochondrial fusion helps maintain mitochondrial continuity and function by connecting two mitochondria and exchanging their contents, while mitochondrial fission allows for quality control of mitochondria by splitting off damaged or dysfunctional mitochondria and targeting them for autophagy for clearance. In cardiomyocytes, the balance of mitochondrial fusion and fission is essential for adapting to the high energy demands of the heart and maintaining cardiac function. In myocardial ischemia-reperfusion injury, the dynamic balance of mitochondria may be disrupted, leading to mitochondrial dysfunction, which in turn exacerbates myocardial injury.

[0005] At present, clinical treatment strategies for myocardial ischemia-reperfusion injury include the use of antioxidants, calcium channel blockers, beta blockers and other drugs, aiming to reduce damage during reperfusion. In addition, researchers are also exploring new treatments, such as regulating mitochondrial function and targeting inflammatory pathways, in order to provide more effective treatments.

[0006] Aiweixin oral liquid (AWX) is a compound oral preparation made from 15 medicinal materials including silkworm cocoons, fragrant blue orchid, musk, etc. (National Medicine Standard Z65020005), which has the effects of dredging collaterals and promoting blood circulation, strengthening the heart and liver, improving microcirculation, calming the nerves, etc. It is included in the "Pharmaceutical Standards of the Ministry of Health of the People's Republic of China·Uyghur Medicine Volume". It is mainly used in the clinic to treat various cardiovascular and cerebrovascular diseases such as chest pain, palpitation and shortness of breath, dizziness, neurasthenia, etc., and the treatment effect has been clinically verified. Summary of the invention

[0007] The purpose of the present invention is to provide the use of Aivxin for preventing or treating mitochondrial abnormality diseases.

[0008] In the first aspect, the present invention claims protection for the use of Aiweixin oral liquid in the preparation of a product for improving mitochondrial homeostasis imbalance. Furthermore, the mitochondrial homeostasis imbalance is caused by cardiovascular disease. In an embodiment of the present invention, the cardiovascular disease is myocardial ischemia-reperfusion injury or ventricular remodeling.

[0009] In a specific implementation case of the present invention, the improvement is a preventive improvement, that is, the medication is taken before the disease occurs.

[0010] Wherein, the product may be a medicine.

[0011] In the second aspect, the present invention claims to protect the use of Aiweixin oral liquid in the preparation of a product for regulating the dynamic balance of fusion and fission of mitochondria in the body or cells.

[0012] Furthermore, the organism is an organism suffering from cardiovascular disease; the cells are cardiovascular disease cell models or isolated myocardial cells from an organism suffering from cardiovascular disease. In an embodiment of the present invention, the cardiovascular disease is myocardial ischemia-reperfusion injury or ventricular remodeling or myocardial cell hypertrophy.

[0013] In a specific implementation case of the present invention, the regulation is preventive regulation, that is, the medication is taken before the disease occurs.

[0014] Wherein, the product may be a medicine.

[0015] In a third aspect, the present invention claims the use of Aiweixin oral liquid in the preparation of a product for reversing the increased expression of mitochondrial fission factors and / or decreased expression of mitochondrial fusion-related factors in myocardial cells caused by cardiovascular disease. In an embodiment of the present invention, the cardiovascular disease is myocardial ischemia-reperfusion injury or ventricular remodeling or myocardial cell hypertrophy.

[0016] Furthermore, the mitochondrial fission factor is MFF and / or FIS1; the mitochondrial fusion-related factor is MFN1 and / or MFN2.

[0017] In a specific implementation case of the present invention, the torsion is a preventive torsion, that is, the medication is taken before the disease occurs.

[0018] Wherein, the product may be a medicine.

[0019] In a fourth aspect, the present invention claims protection for the use of Aiweixin oral liquid in the preparation of products for preventing and / or treating cardiovascular diseases.

[0020] Furthermore, the application is the use of Aiweixin oral liquid as a regulator of the dynamic balance of mitochondrial fusion and fission in the preparation of products for preventing and / or treating cardiovascular diseases.

[0021] In one embodiment of the present invention, the cardiovascular disease is myocardial ischemia-reperfusion injury, ventricular remodeling, or myocardial cell hypertrophy.

[0022] Wherein, the product may be a medicine.

[0023] In a fifth aspect, the present invention claims the use of Aiweixin oral liquid as a positive control drug in screening drugs for preventing and / or treating cardiovascular diseases by regulating the dynamic balance of mitochondrial fusion and fission in the body or cells. In an embodiment of the present invention, the cardiovascular disease is myocardial ischemia-reperfusion injury or ventricular remodeling or myocardial cell hypertrophy.

[0024] In a sixth aspect, the present invention claims a method for screening a drug for preventing and / or treating cardiovascular disease by regulating the dynamic balance of mitochondrial fusion and fission in the body. In an embodiment of the present invention, the cardiovascular disease is myocardial ischemia-reperfusion injury or ventricular remodeling or myocardial cell hypertrophy.

[0025] The method claimed in the present invention for screening drugs for preventing and / or treating cardiovascular diseases by regulating the dynamic balance of mitochondrial fusion and fission in the body may include the following steps: (1) Set up the following groups: Test drug treatment + modeling group: After the test drug is administered to the test animals, cardiovascular disease modeling is performed; Modeling group: The test animals were not given any drugs and were directly subjected to cardiovascular disease modeling; Negative control group: the test animals were not given any drugs and no cardiovascular disease modeling was performed; Positive control drug treatment + modeling group: After administering Aiweixin oral liquid to the test animals, cardiovascular disease modeling was performed; (2) Testing and result determination: If compared with the negative control group, the expression of mitochondrial fission factor in the myocardial tissue of the animals in the modeling group is increased and / or the expression of mitochondrial fusion-related factor is decreased, and compared with the modeling group, the expression of mitochondrial fission factor in the myocardial tissue of the animals in the positive control drug treatment + modeling group is decreased and / or the expression of mitochondrial fusion-related factor is increased, then it is determined as follows whether the drug to be tested is a drug that can prevent and / or treat cardiovascular diseases by regulating the dynamic balance of mitochondrial fusion and fission in the body: If compared with the modeling group, the expression level of the mitochondrial fission factor in the animal myocardial tissue of the test drug treatment + modeling group is decreased and / or the expression level of the mitochondrial fusion-related factor is increased, then the test drug is or is a candidate for the target drug to be screened (i.e., a drug that can prevent and / or treat cardiovascular diseases by regulating the dynamic balance of mitochondrial fusion and fission in the body); otherwise, the test drug is not or is not a candidate for the target drug to be screened.

[0026] Furthermore, the mitochondrial fission factor is MFF and / or FIS1; the mitochondrial fusion-related factor is MFN1 and / or MFN2.

[0027] In the present invention, the regulation of the dynamic balance of mitochondrial fusion and fission in the body or cells described in the above-mentioned related aspects is mainly achieved by regulating the expression level of mitochondrial fission factors and / or mitochondrial fusion-related factors in cardiomyocytes, thereby maintaining mitochondrial homeostasis.

[0028] The Aiweixin oral liquid of the present invention has a significant cardiovascular protective effect, high safety and low side effects; its raw materials are easy to obtain and can be produced by biological fermentation technology, or extracted from natural medicinal materials containing specific ingredients, or prepared by chemical synthesis methods. Aiweixin oral liquid provides a new treatment option for the prevention, diagnosis, monitoring, protection, treatment and research of cardiovascular diseases, has a good application prospect, can be quickly popularized and bring significant social and economic benefits in a relatively short period of time.

[0029] The experiment of the present invention proves that Aiweixin oral liquid has a potential protective effect on cardiovascular diseases, and the protective effect is exerted by regulating the dynamic balance of mitochondrial fusion and fission process. The present invention is of great significance for further understanding the protective mechanism of Aiweixin oral liquid on cardiovascular diseases and improving the corresponding therapeutic effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Electrocardiogram of rats Figure II Lead changes. A is the electrocardiogram of rats before ligation; B is the electrocardiogram of rats after ligation.

[0031] Figure 2 The effect of different doses of Aiweixin oral solution on myocardial infarction area in rats. The left picture shows the results of myocardial TTC staining; the right picture shows the statistical chart of myocardial TTC staining results. n=6, #P<0.05 vs SHAM group; *P<0.05, vs I / R group.

[0032] Figure 3 The effect of different doses of Aiweixin oral liquid on myocardial tissue pathological damage in rats of different groups (HE staining).

[0033] Figure 4 The effect of different doses of Aiweixin oral solution on the levels of myocardial injury markers in the serum of rats in each group. n=6, #P<0.05 vs SHAM group; *P<0.05, vs I / R group.

[0034] Figure 5 The effect of different doses of Aiweixin oral solution on the oxidative stress indexes in the serum of rats in each group. n=6, #P<0.05 vs SHAM group; *P<0.05, vs I / R group.

[0035] Figure 6 The effect of different doses of Aiweixin oral solution on the expression levels of MNF1, MNF2, MFF and FIS1 proteins in the myocardial tissue of rats in each group. n=6, #P<0.05 vs SHAM group; *P<0.05, vs I / R group.

[0036] Figure 7 Application of Aiweixin oral liquid in preventing and treating ventricular remodeling / cardiomyocyte hypertrophy. #P<0.05 vs Control group; *P<0.05, vs M group. DETAILED DESCRIPTION

[0037] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0038] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0039] Example 1: Application of Aiweixin oral liquid in the preparation of drugs for improving mitochondrial quality control and preventing and treating myocardial ischemia-reperfusion 1. Experimental Methods 1. Establishment of myocardial ischemia-reperfusion model 72 healthy male Wistar rats were provided by the Animal Experiment Center of Xinjiang Medical University, with a body weight of 200-250 g. Before the experiment, the rats needed to fast for 12 hours but had free access to water to ensure the safety of the animals during the experiment and the reliability of the experimental results. After being weighed, the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (0.3 mL / 100 g body weight) and fixed on the operating table in a supine position. The surgical area included the neck and chest, which needed to be prepared, disinfected, and covered with a sterile drape. The electrocardiogram monitoring system was connected to record the rats' electrocardiogram activity and exclude individuals with abnormal electrocardiograms. Through a tracheotomy in the neck, an artificial respirator was connected for assisted breathing (tidal volume was set to 5 mL / 100 g, respiratory rate was 60 to 80 times / min, respiratory ratio was 2:1, and positive end-expiratory pressure was applied).

[0040] The skin was cut longitudinally about 3 cm between the 3rd and 4th ribs 0.5 cm to the left of the sternum, and the subcutaneous tissue and the pectoralis major muscle to the intercostal muscle were bluntly separated and fixed with hemostats. The 3rd and 4th ribs were cut open, the chest cavity was opened, and the heart was exposed. The left coronary vein trunk and the accompanying left anterior descending coronary artery (LAD) were found between the lower edge of the left atrial appendage and the pulmonary artery cone. After squeezing out the heart, the aortic root was ligated about 2 mm between the lower edge of the left atrial appendage and the pulmonary artery cone. The ligature was passed through the left coronary vein trunk, and a small plastic tube was put on the line to fix it. After ligation, the color change of the anterior wall of the left ventricle and the elevation of the ST segment of the electrocardiogram indicated myocardial ischemia. After 30 minutes, the ligature was cut to achieve reperfusion, at which time the ST segment decreased by about 50%. After the model was established, the chest cavity, muscles and skin were sutured layer by layer and disinfected. Finally, penicillin was injected to prevent postoperative infection. 90 minutes after the coronary artery ligation surgery, the rats were anesthetized again, and relevant indicators were tested immediately, and arterial blood and heart were collected.

[0041] 2. Experimental grouping and treatment of myocardial ischemia-reperfusion model group A total of 72 healthy male Wistar rats were randomly divided into sham operation group, model group, AWX low-dose administration group (1 ml kg -1 ), AWX medium dose group (2 ml·kg -1 ), AWX high-dose group (4 ml·kg -1 ), trimetazidine group (10 mg kg -1 ), 12 in each group.

[0042] Drug administration group (I / R+AWX 1ml·kg -1 ): Rats were given Aiweixin oral solution (1 ml kg -1 ) were intragastrically intervened for 10 days, and the myocardial ischemia-reperfusion model was prepared using the method in step 1.

[0043] Drug administration group (I / R+AWX 2ml·kg -1 ): Rats were given Aiweixin oral solution (2ml·kg -1 ) were intragastrically intervened for 10 days, and the myocardial ischemia-reperfusion model was prepared using the method in step 1.

[0044] Drug administration group (I / R+AWX 4ml·kg -1 ): Rats were given Aiweixin oral solution (4ml·kg -1 ) were intragastrically intervened for 10 days, and the myocardial ischemia-reperfusion model was prepared using the method in step 1.

[0045] Positive control group (I / R+TMZ): Rats were given trimetazidine (10 mg kg -1 ) were intragastrically intervened for 10 days, and the myocardial ischemia-reperfusion model was prepared using the method in step 1.

[0046] Sham group: Rats were given normal saline (1 ml kg -1 ) The patients were given oral gavage for 10 days, and after thoracotomy, only needles were inserted and threaded, but no ligation was performed.

[0047] Model group (I / R): rats were given normal saline (1 ml kg -1 ) were intragastrically intervened for 10 days, and the myocardial ischemia-reperfusion model was prepared using the method in step 1.

[0048] 3. Animal handling In the experiment, rats were dissected to remove the heart, and then the heart was washed with saline solution to remove congestion. The experiment involved 6 rats to determine the area of ​​myocardial infarction. In the operation, the rat's heart was first removed, and the atrial tissue and excess fat were removed and weighed. Then, the heart of each group of 6 rats was dissected and separated, and cut into 5 slices along the coronal plane. TTC stain was stained in a 37 ℃ water bath in the dark for 30 minutes, and then fixed with 4% paraformaldehyde and observed. Normal myocardial tissue is red, and the tissue in the infarction area is pale white. After taking pictures, the images were analyzed with Imagepro software to detect the area of ​​myocardial infarction (myocardial infarction area = total heart infarction area / total heart slice area). About 2 mL of blood samples were collected from the remaining 6 rats in each group through the carotid artery, and then centrifuged at 3000 rpm for 10 minutes to obtain the supernatant, which was stored in a -80°C refrigerator for subsequent use. After the heart was perfused with PBS, the rat heart was completely separated, the total weight of the heart was weighed, and the heart tissue was immediately frozen in liquid nitrogen after sampling. Later, it was transferred to cryopreservation at -80°C. The samples were mainly used for RNA extraction, protein extraction, etc. 4. Electrocardiogram After anesthesia, the rats in each group were immediately depilated and fixed, and connected to a ventilator after endotracheal intubation. Electrodes were placed at the ends of the rats' limbs to open the chest for modeling. The standard II lead electrocardiogram of the rats was observed and recorded in real time.

[0049] 5. TTC staining The heart tissue was quickly removed, and 6 slices of about 2 mm were cut from the apex to the base of the heart. They were placed in 2% TTC solution and stained at 37 °C in the dark for 20 min. After staining, they were fixed with 4% paraformaldehyde for 24 h and then photographed. The red area is normal myocardial tissue, and the white area is ischemic myocardium. Image J software was used to analyze and calculate the area of ​​ischemic infarction and non-infarction areas. 2,3,5-triphenyltetrazolium chloride (TTC) staining solution was a product of Beijing Solebow Technology Co., Ltd.

[0050] 6. HE staining Five heart tissues fixed in 4% paraformaldehyde were randomly selected from each group. Excess water was removed with filter paper, and the tissues were embedded in paraffin and made into 4 μm thick sections. The sections were stained and sealed according to the instructions of hematoxylin-eosin (HE) staining. Changes in myocardial tissue were observed under a microscope (400 times) and images were collected.

[0051] 7. Detection of myocardial injury markers The prepared serum samples were added to determine the levels of LDH and cTnI according to the instructions of the kit (Beijing Solarbio, Beijing, China). The lactate dehydrogenase (LDH) activity detection kit was a product of Beijing Solarbio Technology Co., Ltd. The rat cardiac troponin I (cTnI) content detection kit was a product of Wuhan FineTest Co., Ltd.

[0052] 8. Oxidative stress detection The operation was carried out according to the instructions of the kit, and the prepared serum samples were added to determine the levels of SOD and MDA. The rat superoxide dismutase (SOD) activity detection kit was a product of Nanjing Jiancheng Bioengineering Institute. The rat malondialdehyde (MDA) content detection kit was a product of Beijing Solebow Technology Co., Ltd.

[0053] 9. Detection of MNF1, MNF2, MFF, and FIS1 protein expression levels in rat myocardial tissue by Western blot Proteins in myocardial tissue were extracted by conventional methods, and the protein samples were denatured at high temperature after BCA protein quantification. SDS-PAGE gel electrophoresis was used and transferred to PVDF membrane, which was blocked with 5% skim milk at room temperature for 2 h and incubated with primary antibody at 4 °C overnight. The second antibody was incubated at room temperature for 1 h the next day, and then the PVDF membrane was placed in an exposure instrument and ECL developer was added to develop. Finally, Image J software was used to analyze the gray value of each group of bands, and the protein expression was recorded and counted. Anti-mitochondrial fission factor (MFF) antibody and anti-mitochondrial fission protein 1 (FIS1) antibody (Wuhan Sanying Biotechnology Co., Ltd., catalog numbers 17090-1-AP and 10956-1-AP, respectively); anti-mitochondrial outer membrane fusion protein 1 (MFN1) antibody and anti-mitochondrial outer membrane fusion protein 2 (MFN2) antibody (Abcam, UK, catalog numbers AB221661 and AB124773, respectively).

[0054] 10. Statistical processing GraphPad Prism 8.0 software was used to perform statistical analysis of the data. ) indicated that one-way ANOVA was used for comparison among groups, and LSD-t test was used for pairwise comparison.

[0055] 2. Results and Analysis 1. Rat ECG Figure II Lead changes In the myocardial ischemia-reperfusion injury (MIRI) model group, low-dose Avixin oral liquid group, medium-dose Avixin oral liquid group, high-dose Avixin oral liquid group, and positive control drug trimetazidine group, all rats underwent electrocardiogram monitoring. When ST segment elevation was observed in the electrocardiogram, it indicated that the rat myocardial ischemia model was successfully established. For detailed results, see Figure 1 .

[0056] 2. Effects of different doses of Aiweixin oral solution on myocardial infarction area in rats TTC staining showed that the myocardial infarction area of ​​rats in the myocardial ischemia-reperfusion injury group (MIRI) was significantly enlarged compared with the sham group; and compared with the MIRI group, the groups treated with low, medium and high doses of Avixin oral liquid and the group using the positive control drug trimetazidine showed significantly reduced myocardial infarction area. For details, see Figure 2 .

[0057] 3. Effects of different doses of Aiweixin oral liquid on myocardial tissue pathological damage in rats of different groups Hematoxylin-eosin (HE) staining results showed that compared with the sham operation group (Sham), the myocardial cells of rats in the myocardial ischemia-reperfusion injury group (MIRI) showed pathological changes such as nuclear fragmentation, nuclear dissolution, inflammatory cell infiltration, interstitial edema and blurred muscle fiber boundaries. Compared with the MIRI group, the disordered arrangement of myocardial cells and the degree of myocardial injury were alleviated in the groups treated with low, medium and high doses of Avixin oral liquid and the group treated with the positive control drug trimetazidine. For detailed results, see Figure 3 .

[0058] 4. Effects of different doses of Aiweixin oral liquid on the levels of myocardial injury markers in the serum of rats in each group The results of myocardial injury biomarkers showed that compared with the sham group (Sham), the levels of lactate dehydrogenase (LDH) and cardiac troponin I (cTnI) in the serum of rats in the myocardial ischemia-reperfusion injury group (MIRI) were significantly increased. Compared with the MIRI group, the levels of LDH and cTnI in the serum of the groups treated with low, medium and high doses of Avixin oral liquid and the group using the positive control drug trimetazidine were significantly reduced. For details, see Figure 4 .

[0059] 5. Effects of different doses of Aiweixin oral liquid on oxidative stress indicators in the serum of rats in each group.

[0060] The results of oxidative stress indicators showed that compared with the sham operation group (Sham), the superoxide dismutase (SOD) activity in the serum of rats in the myocardial ischemia-reperfusion injury group (MIRI) was reduced; compared with the MIRI group, the SOD activity in the serum of rats in the groups treated with low and high doses of Avixin oral liquid and the group treated with the positive control drug trimetazidine increased. In addition, compared with the Sham group, the malondialdehyde (MDA) level in the serum of rats in the MIRI group increased; compared with the MIRI group, the MDA content in the groups treated with low, medium, and high doses of Avixin oral liquid or trimetazidine was significantly reduced. For detailed results, see Figure 5 .

[0061] 6. Effects of different doses of Aiweixin oral liquid on the expression levels of MNF1, MNF2, MFF, and FIS1 proteins in myocardial tissue of rats in each group Western blot results showed that compared with the sham operation group (Sham), the protein expression levels of mitochondrial fission factors MFF and FIS1 in the myocardial tissue of rats in the myocardial ischemia-reperfusion injury group (MIRI) were significantly increased; the protein expression levels of mitochondrial fusion-related factors MFN1 (Mitofusin 1) and MFN2 (Mitofusin 2) in the myocardial tissue were significantly decreased. Compared with the MIRI group, the groups treated with low, medium and high doses of Aivoxil oral liquid and the positive control drug trimetazidine group showed up-regulated expression of mitochondrial fusion proteins MFN1 and MFN2, and these drug-treated groups also showed down-regulated expression of mitochondrial fission proteins MFF and FIS1. These changes were statistically significant, indicating that Aivoxil oral liquid exerts its protective effect against myocardial ischemia-reperfusion injury by regulating the expression of mitochondrial fission and fusion-related proteins (i.e., by regulating the dynamic balance of mitochondrial fusion and fission). For detailed results, see Figure 6 .

[0062] Example 2: Application of Aiweixin oral liquid in preventing and treating ventricular remodeling / cardiomyocyte hypertrophy In this example, a ventricular remodeling / myocardial mast cell model was constructed using H9C2 cardiomyocytes, and then the pharmacodynamic effects of AWX oral solution on it were studied. The details are as follows: H9C2 cells were seeded in six-well plates at a density of 2 × 10 5 The cells were cultured in DMEM containing 10% FBS for one day. When the cell confluence reached 80%, the cells were starved with DMEM containing 0.5% FBS for 24 h. Then, the cells were incubated with AngⅡ (angiotensin II) prepared in DMEM containing 10% FBS and different concentrations of AWX for 48 h. Afterwards, RNA was extracted to detect the gene expression of anti-apoptotic protein (Bcl2) and pro-apoptotic protein (Bax). The dosage of AngⅡ was 100mmol / L (final concentration).

[0063] AWX administration dose: low-dose group (L group) 50μl AWX / 1ml culture medium; high-dose group (H group) 100μl AWX / 1ml culture medium; model group (M group) no AWX. Each concentration is the final drug concentration at the time of treatment.

[0064] In the experiment, H9C2 cells without any intervention were set up as a control group (Control group).

[0065] The results are as follows Figure 7 As shown: Compared with normal H9C2 cells (Control group), the Bax / Bcl2 ratio in the model group (M group) cells was increased, indicating that under the stimulation of AngII, the apoptotic tendency of the model group cells was enhanced, which may be due to the upregulation of Bax and the downregulation of Bcl2, leading to the activation of the cell apoptosis pathway, thereby promoting cell apoptosis.

[0066] Compared with the model group (group M), the Bax / Bcl2 ratio in the AWX low-dose group (group L) and high-dose group (group H) decreased, and with the increase of AWX dosage, the Bax / Bcl2 ratio decreased more significantly, indicating that AWX may reduce the apoptotic response induced by AngII by regulating the expression of apoptosis-related factors. With the increase of dosage, the effect of AWX may be more significant, further reducing the apoptotic tendency of cells, suggesting that AWX may have a certain protective effect. Mitochondrial homeostasis imbalance can cause cell damage and induce cell apoptosis. Improving mitochondrial homeostasis can promote the restoration of normal mitochondrial function, and cells can better cope with stress and damage, thereby reducing apoptosis.

[0067] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principles of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departure from the disclosed scope in the application, and changes made with conventional techniques known in the art.

Claims

1. Application of Aiweixin oral liquid in the preparation of products for improving mitochondrial homeostasis imbalance.

2. The use according to claim 1, characterized in that: The imbalance of mitochondrial homeostasis is caused by cardiovascular diseases.

3. The use of Aiweixin oral liquid in the preparation of products for regulating the dynamic balance of fusion and fission of mitochondria in the body or cells.

4. The use according to claim 3, characterized in that: The organism is an organism suffering from cardiovascular disease; the cells are a cardiovascular disease cell model or isolated myocardial cells from an organism suffering from cardiovascular disease.

5. Use of Aiweixin oral liquid in the preparation of a product for reversing the increased expression of mitochondrial fission factors and / or the decreased expression of mitochondrial fusion-related factors in cardiomyocytes caused by cardiovascular diseases.

6. The use according to claim 5, characterized in that: The mitochondrial fission factor is MFF and / or FIS1; the mitochondrial fusion-related factor is MFN1 and / or MFN2.

7. Use of Aiweixin oral liquid in the preparation of products for preventing and / or treating cardiovascular diseases; Furthermore, the application is the use of Aiweixin oral liquid as a regulator of the dynamic balance of mitochondrial fusion and fission in the preparation of products for preventing and / or treating cardiovascular diseases.

8. Use of Aiweixin oral liquid as a positive control drug in screening drugs for preventing and / or treating cardiovascular diseases by regulating the dynamic balance of mitochondrial fusion and fission in the body or cells.

9. A method for screening drugs for preventing and / or treating cardiovascular diseases by regulating the dynamic balance of mitochondrial fusion and fission in the body, comprising the following steps: (1) Set up the following groups: Test drug treatment + modeling group: After the test drug is administered to the test animals, cardiovascular disease modeling is performed; Modeling group: The test animals were not given any drugs and were directly subjected to cardiovascular disease modeling; Negative control group: the test animals were not given any drugs and no cardiovascular disease modeling was performed; Positive control drug treatment + modeling group: After administering Aiweixin oral liquid to the test animals, cardiovascular disease modeling was performed; (2) Testing and result determination: If compared with the negative control group, the expression of mitochondrial fission factor in the myocardial tissue of the animals in the modeling group is increased and / or the expression of mitochondrial fusion-related factor is decreased, and compared with the modeling group, the expression of mitochondrial fission factor in the myocardial tissue of the animals in the positive control drug treatment + modeling group is decreased and / or the expression of mitochondrial fusion-related factor is increased, then it is determined as follows whether the drug to be tested is a drug that can prevent and / or treat cardiovascular diseases by regulating the dynamic balance of mitochondrial fusion and fission in the body: If compared with the modeling group, the expression level of the mitochondrial fission factor in the animal myocardial tissue of the test drug treatment + modeling group is decreased and / or the expression level of the mitochondrial fusion-related factor is increased, then the test drug is or is a candidate for the target drug to be screened; otherwise, the test drug is not or is not a candidate for the target drug to be screened.

10. The method according to claim 9, characterized in that: The mitochondrial fission factor is MFF and / or FIS1; the mitochondrial fusion-related factor is MFN1 and / or MFN2.

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