Application of RNF130 activator in preparation of medicine for preventing and / or treating ischemic heart disease

Abstract

The invention discloses application of an RNF130 activator in preparation of a medicine for preventing and / or treating ischemic heart disease, and belongs to the technical field of biological medicine. Research results show that when an RNF130 activator is given in a mouse of a myocardial ischemia reperfusion injury model, the left ventricular ejection fraction and the left ventricular short-axis shortening rate of the mouse can be improved, myocardial tissue pathological injury can be relieved, the protein expression levels of NOX-2, BAX and TNF-alpha are reduced, the myocardial infarction area is reduced, and myocardial cell apoptosis is reduced; therefore, the RNF130 activator has prevention and treatment effects on the myocardial ischemia-reperfusion injury, that is, the RNF130 activator can be used as an effective component of the medicine for preventing or treating diabetes complicated by myocardial ischemia-reperfusion injury, and a new strategy is provided for preparation of the medicine for preventing and treating myocardial ischemia-reperfusion injury.

Description

Technical Field

[0001] The present invention relates to application of an RNF130 activator in preparing a medicine for preventing and / or treating ischemic heart disease, and belongs to the technical field of biomedicine. Background Art

[0002] Myocardial ischemia-reperfusion injury is a condition in which the cell metabolism dysfunction and tissue structure damage that occur when the blood supply to the ischemic myocardium is restored by reperfusion therapy lead to regional myocardial ischemia and eventually cause myocardial necrosis. This disease is characterized by myocardial infarction, accompanied by decreased ventricular systolic function, reduced left ventricular ejection fraction and left ventricular short-axis shortening. The disease has an acute onset and death can occur at any stage of the disease.

[0003] Currently, some drugs are being studied to reduce myocardial ischemia-reperfusion injury, such as antioxidants, inflammation inhibitors, and apoptosis inhibitors. Myocardial protection is to protect the myocardium by reducing cell damage caused by myocardial ischemia-reperfusion. These procedures include coronary artery bypass grafting, myocardial hormone injection, and cryocooling. Researchers are developing new monitoring methods to assess the extent of myocardial reperfusion injury and predict the prognosis of patients. These methods include myocardial enzyme marker detection, myocardial imaging, and biomarker analysis. Methods to prevent myocardial reperfusion injury include improving preoperative management, reducing myocardial ischemia time, and increasing myocardial blood flow and oxygenation levels. Researchers are exploring comprehensive treatment strategies that combine drug therapy, myocardial protection, and reperfusion injury monitoring to minimize the damage caused by myocardial ischemia-reperfusion.

[0004] In the prior art, patent CN117815396A provides a drug that targets the UCP2 gene to treat myocardial ischemia-reperfusion injury, which alleviates myocardial ischemia-reperfusion injury by inhibiting myocardial cell ferroptosis; patent CN117138044A provides the use of Tmprss13 as a target for the treatment of myocardial ischemia-reperfusion injury, that is, supplementing Tmprss13 protein in vivo can alleviate myocardial ischemia-reperfusion injury by improving myocardial cell apoptosis and necrosis, pathological ventricular remodeling, and reducing cardiac fibrosis. However, overall, the current therapeutic targets for the prevention and treatment of myocardial ischemia-reperfusion injury cannot meet clinical needs, and new therapeutic targets need to be found.

[0005] RNF130 (Ring Finger Protein 130) is the mammalian homolog of the Drosophila protease-associated domain E3 ligase and a protein with E3 ubiquitin ligase activity. It plays a role in a variety of biological processes, including regulating apoptosis, cell cycle, and signal transduction. RNF130 can control the selective degradation of proteins by proteasomes, and can also catalyze the transfer of ubiquitin to target proteins, rapidly modify and regulate target proteins, thereby conferring specificity to target proteins, modifying proteins, and regulating their cellular localization. Today, RNF130 can be used for the treatment of pulmonary fibrosis, regulation of blood lipid levels, and drug development for autoimmune diseases, but at present, there has been no relevant research and reports on the role and mechanism of RNF130 in myocardial ischemia-reperfusion injury. Summary of the invention

[0006] In order to solve the above technical problems, the present invention provides an application of a RNF130 activator in the preparation of a drug for preventing or treating myocardial ischemia-reperfusion injury;

[0007] The present invention found that RNF130 expression was reduced in the heart tissue of mice with myocardial ischemia-reperfusion injury, which may be related to the development of the disease and may become an effective target for the treatment of myocardial ischemia-reperfusion injury.

[0008] The first object of the present invention is to provide the use of RNF130 as a therapeutic target in the preparation of a drug for ischemic heart disease, wherein the drug increases the expression of RNF130 gene or protein.

[0009] In one embodiment, the medicament comprises an activator of RNF130.

[0010] In one embodiment, the activator includes an agent that enhances RNF130 gene expression and / or an agent that enhances RNF130 protein expression.

[0011] In one embodiment, the drug improves myocardial damage induced by cardiac ischemia-reperfusion injury, improves pathological changes in cardiac tissue; reduces the expression levels of myocardial infarction markers c-Tnl, 1L-1β and 1L-6; and inhibits the expression of inflammation and oxidative damage markers NOX-2, BAX, and TNF-α proteins.

[0012] The second object of the present invention is to provide use of an RNF130 activator in preparing a medicament for treating and / or alleviating cardiac ischemia-reperfusion injury.

[0013] In one embodiment, the RNF130 activator includes an agent that enhances RNF130 gene expression and / or an agent that enhances RNF130 protein expression.

[0014] The third object of the present invention is to provide the use of RNF130 gene or protein in screening drugs for preventing or treating cardiac ischemia-reperfusion injury.

[0015] In one embodiment, the application includes using the expression of the RNF130 gene or protein to verify whether a drug can be used to prevent or treat cardiac ischemia-reperfusion injury.

[0016] In one embodiment, the drug for preventing or treating cardiac ischemia-reperfusion injury can increase the expression of RNF130 gene or protein.

[0017] The fourth object of the present invention is to provide a drug capable of preventing or treating cardiac ischemia-reperfusion injury.

[0018] In one embodiment, the drug comprises an activator of the RNF130 gene and / or its expression product.

[0019] In one embodiment, the drug is a vector for gene therapy, including vectors for gene editing, gene expression regulation or gene delivery; or the drug is a vector for cell therapy, including stem cells, immune cells or repair cells.

[0020] In one embodiment, the medicament prevents or treats diabetic cardiomyopathy by increasing the expression of RNF130 gene or protein.

[0021] In one embodiment, the drug contains the above-mentioned RNF130 activator and a pharmaceutically acceptable excipient.

[0022] In one embodiment, the auxiliary material includes any one of a carrier, a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an emulsifier, a solubilizer, a solubilizing agent, an osmotic pressure regulator, a surfactant, a coating material, a colorant, a pH regulator, an antioxidant, an antibacterial agent or a buffer, or a combination of at least two thereof.

[0023] In one embodiment, the carrier is one or more of a virus, a liposome, and a nanoparticle.

[0024] In one embodiment, the excipient is one or more of mannitol, lactose, fatty acid, and polyethylene glycol.

[0025] In one embodiment, the dosage form of the drug includes any one of suspension, granules, capsules, powders, tablets, emulsions, solutions, pills, injections, suppositories, enemas, aerosols, patches or drops.

[0026] In one embodiment, the administration route of the drug includes oral administration, sublingual administration, rectal administration, skin mucosal administration, inhalation administration or injection administration.

[0027] In one embodiment, the drug of the present invention has at least one or more of the following functions:

[0028] (1) Restore the reduction of left ventricular ejection fraction and left ventricular fractional shortening caused by cardiac ischemia-reperfusion injury, reverse cardiac function damage, and improve cardiac tissue pathological changes;

[0029] (2) reduce the expression levels of myocardial infarction markers c-Tnl, 1L-1β, and 1L-6;

[0030] (3) Inhibit the expression of inflammation and oxidative damage markers such as NOX-2, BAX, and TNF-α proteins.

[0031] The present invention finds that injecting RNF130 overexpression plasmid into mice can increase the expression of RNF130, improve the cardiac function of mice with myocardial ischemia-reperfusion injury, reduce myocardial infarction markers, inhibit the expression of inflammation and oxidative damage markers NOX-2, BAX, and TNF-α proteins, and reduce the area of ​​myocardial infarction, thereby preventing and treating myocardial ischemia-reperfusion injury and providing a new target for its treatment.

[0032] Beneficial Effects

[0033] RNF130 activators can improve the cardiac function of mice with myocardial ischemia-reperfusion injury, reverse the decrease in left ventricular ejection fraction (EF) and left ventricular fractional shortening (FS) caused by myocardial ischemia-reperfusion injury, reduce cardiomyocyte apoptosis, reduce the levels of myocardial infarction markers c-Tnl, 1L-1β and 1L-6, inhibit the expression of inflammation and oxidative damage markers NOX-2, BAX, and TNF-α proteins, and reduce the area of ​​myocardial infarction, thereby preventing and treating myocardial ischemia-reperfusion injury and providing new targets for the preparation of drugs for the prevention and treatment of myocardial ischemia-reperfusion injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 To investigate the effect of overexpression of RNF130 on cardiac cell apoptosis in mice;

[0035] Figure 2 To investigate the effect of knocking out RNF130 on cardiac cell apoptosis in mice;

[0036] Figure 3The effect of RNF130 activator on improving cardiac function in mice with myocardial ischemia-reperfusion injury; (A) Left ventricular ejection fraction (EF); (B) Left ventricular fractional shortening (FS); (C) Serum levels of myocardial infarction markers c-Tnl, 1L-1β and 1L-6; (D) Transthoracic echocardiography results in mice. *p<0.05vs.Sham, vs.MI / RI+Vector.

[0037] Figure 4 The effect of RNF130 activator on improving myocardial ischemia-reperfusion injury in mice; (A) IHC staining of heart sections; (B) DHE staining of heart sections; (C) TUNEL staining of heart sections;

[0038] Figure 5 The effect of using RNF130 activator to improve the myocardial infarction level in mice with myocardial ischemia-reperfusion injury; (A) Evans blue & TTC staining; (B) Myocardial infarction area statistics; *p < 0.05 vs. Sham, vs.MI / RI;

[0039] Figure 6 is the fluorescence intensity of F4 / 80 (macrophage-specific marker) in mouse heart. DETAILED DESCRIPTION

[0040] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explaining the present invention and are not used to limit the present invention.

[0041] Experimental methods:

[0042] 1. Treatment methods of cardiac ischemia-reperfusion model group (MI / RI group):

[0043] Male C57BL / 6J mice aged 6 to 8 weeks were purchased from Nanjing Sibeifu Biotechnology Co., Ltd.;

[0044] The in vivo mouse cardiac ischemia-reperfusion model can be established when C57BL / 6 mice grow to 6 to 8 weeks of age. After the mice are anesthetized with 2% isoflurane gas, endotracheal intubation is performed and respiratory support is provided by a ventilator; then a small skin incision (about 1.2 cm) is made on the left chest, and the pectoralis major and pectoralis minor muscles are cut to expose the fourth intercostal space. A small hole is made in the fourth intercostal space with a needle holder to open the pleura and pericardium. With the needle holder slightly stretching the intercostal space, the heart is smoothly and gently "popped out" from the hole. Use 6-0 silk thread to locate the left coronary artery (LCA) and insert the needle at a position about 3 mm from its upper end and tie a slipknot ligature. When the anterior wall of the left ventricle becomes pale, the ligation is considered successful. After ligation, the heart is immediately placed back into the chest cavity, and the longer end of the slipknot is left outside the chest. Manual suction is performed to prevent pneumothorax, and the muscles and skin are closed using a purse-string suture. At this time, the half-hour ischemia timer begins. The mice were then allowed to breathe room air and monitored during the recovery period, which was generally completed within 3-5 minutes. The ligature was slowly withdrawn after half an hour, and reperfusion began. Reperfusion lasted for 24 hours. In the sham operation group, the chest cavity was opened and then closed and sutured, and the cardiac MI / R model was not established.

[0045] 2. Mouse serum sample collection:

[0046] (1) Anesthetized mice: Mice were deeply anesthetized with sodium pentobarbital;

[0047] (2) Blood collection: Orbital venous blood sampling was used to collect the blood of mice into centrifuge tubes;

[0048] (3) Separation of serum: The blood sample was allowed to stand at room temperature for 2 hours, and then placed in a 4°C refrigerator overnight. Finally, the sample was centrifuged to separate the serum, and the serum was transferred to a new centrifuge tube for subsequent analysis.

[0049] 3. Mouse heart sample collection:

[0050] (1) Anesthetized mice: Mice were deeply anesthetized with sodium pentobarbital;

[0051] (2) Heart exposure: Perform thoracotomy to expose the mouse heart; be careful not to damage the heart and other organs during the operation;

[0052] (3) Sample collection: Using sterile instruments, carefully remove the heart tissue sample;

[0053] (4) Sample processing: According to research needs, the cardiac tissue samples are processed, such as slicing, lysis or fixation, to maintain the integrity and stability of the samples.

[0054] 4. Transthoracic cardiac ultrasound in mice:

[0055] Transthoracic cardiac ultrasound examination of mice was performed using a Vevo 2100 high-resolution imaging system (Visualsonics, Toronto, ON, Canada) equipped with a 30 MHz probe, referring to the equipment operating manual. The specific steps are as follows:

[0056] (1) Anesthetizing mice: Use appropriate anesthetics (such as isoflurane or pentobarbital) to fully anesthetize the mice and ensure that they are in a pain-free and motionless state;

[0057] (2) Positioning the mouse: Place the mouse on a heating pad and adjust the position so that the chest is exposed to the ultrasound probe;

[0058] (3) Prepare the probe: Connect the 30 MHz probe to the Vevo 2100 high-resolution imaging system and calibrate the probe to ensure good imaging quality;

[0059] (4) Applying gel: Apply a small drop of ultrasound gel on the appropriate position of the mouse's chest to provide an ultrasound transmission medium and reduce air gaps;

[0060] (5) Ultrasound imaging: Gently place the ultrasound probe on the chest area coated with ultrasound gel and fix it in a suitable position. Start the Vevo 2100 system for image acquisition;

[0061] (6) Obtaining images: Using the real-time imaging function provided by the system to obtain the heart image of the mouse;

[0062] (7) Data analysis: importing the collected electrocardiographic data into analysis software to calculate and analyze cardiac function parameters;

[0063] (8) End of operation: After completing image acquisition and data analysis, turn off the Vevo 2100 system, remove the mouse from the heating pad, and restore anesthesia.

[0064] 5. HE staining method:

[0065] Mouse heart HE staining: Mouse heart HE staining is a commonly used tissue staining method used to observe and analyze the morphological structure of mouse heart tissue. The specific operation method is as follows:

[0066] (1) Sample fixation: The mouse heart was removed and fixed with 10% buffered formalin solution to maintain the integrity of the tissue structure;

[0067] (2) Tissue processing: Dehydrate and embed the fixed heart tissue. First, dehydrate the heart tissue by soaking it in alcohol with increasing concentrations to remove water. Then, use xylene to make the dehydrated heart tissue transparent to make the tissue compatible with paraffin.

[0068] (3) Slice preparation: The cleared heart tissue was embedded in paraffin to prepare a paraffin block. Subsequently, the paraffin block was cut into 10 μm thick tissue sections using a microtome and the sections were transferred to glass slides;

[0069] (4) Staining: HE staining is performed on the slices. First, the slices are placed in an alkaline and acidic hydrolysis dye.

[0070] The dye solution contains heme dye and eosin dye. Under alkaline conditions, eosin dye combines with cell nuclear DNA to dye it red; under acidic conditions, heme dye dyes cytoplasm and intercellular matrix to light blue. After a certain period of staining, the slices are transferred to a dehydrating solution to remove excess dye;

[0071] (5) Sealing and observation: Seal the slices with transparent plastic foam and observe under a microscope. In the mouse heart tissue slices after HE staining, the cell nucleus appears red, and the cytoplasm and intercellular matrix appear light blue. The tissue structure, cell morphology, and tissue type can be observed and analyzed under a microscope.

[0072] 6. Isolation and culture of cardiac cells

[0073] Primary cardiomyocytes, endothelial cells, and fibroblasts were isolated from the hearts of newborn Sprague Dawley rats. The specific methods are as follows:

[0074] (1) Animal preparation: Select healthy newborn Sprague Dawley rats;

[0075] (2) Anesthesia and dissection: The rats were fully anesthetized with sodium pentobarbital, and then the hearts were quickly dissected out and cut into approximately 1-2 mm 3 small pieces;

[0076] (3) Cell separation: Use trypsin and collagenase to separate different types of cells;

[0077] a. Isolation of primary cardiomyocytes: Add digestion buffer containing 0.08% trypsin to the heart tissue, incubate at 37°C for 20-30 minutes, and then stop digestion with pre-cooled culture medium. Centrifuge the mixed cell suspension and remove the supernatant, then resuspend the cells and perform washing steps;

[0078] b. Endothelial cell isolation: Add digestion buffer containing 0.1% trypsin and 0.1% collagenase to the heart tissue, incubate at 37°C for 30-40 minutes using an oscillator, and then filter out solid tissue debris through a filter. Centrifuge the cell suspension and remove the supernatant, then resuspend the cells and perform a washing step;

[0079] c. Fibroblast isolation: Add digestion buffer containing 0.2% collagenase and 0.25% trypsin to the heart tissue, incubate at 37°C for 60-90 minutes using an oscillator, and then filter through a filter to remove solid tissue debris. Centrifuge the cell suspension and remove the supernatant, then resuspend the cells and perform a washing step;

[0080] (4) Cell culture: The isolated primary cardiomyocytes, endothelial cells, and fibroblasts were inoculated into culture dishes pre-coated with a suitable matrix (e.g., collagen coating), and provided with appropriate culture medium (e.g., DMEM / F12, DMEM, etc.) and culture conditions (37°C, 5% CO 2 concentration, etc.).

[0081] 7. Myocardial infarction area test method:

[0082] Evans Blue + TTC double staining is a combined technique that can simultaneously observe myocardial cell integrity and metabolic activity. Evans Blue + TTC double staining of the heart is mainly used to evaluate the relative size of the danger zone and the infarct area in the mouse heart ischemia-reperfusion injury model, visualize the degree of damage after heart injury, and thus locate and analyze the myocardial infarction area. The method is as follows:

[0083] (1) Prepare 1.5% Evans Blue dye solution (1.5 g + 100 mL double distilled water) and 1% TTC dye solution for later use.

[0084] (2) Anesthetize the mouse with anesthetic.

[0085] (3) The mouse's chest was opened and the anterior descending branch of the mouse's heart was quickly ligated at the original ligation site using a 6-0 suture needle.

[0086] (4) Carefully clamp the ascending aorta with hemostatic forceps and slowly and carefully inject Evans Blue dye from the proximal end of the clamp. It can be observed that the mouse heart quickly turns blue, and there is no obvious change below the ligature.

[0087] (5) The heart was quickly separated and stored in a -20-80℃ refrigerator.

[0088] (6) After collecting all the hearts according to the above steps, take the hearts out of the refrigerator and slice them layer by layer from the apex with a blade. Each slice is about 2 mm thick, and a total of 4 slices are cut.

[0089] (7) Place the cut heart slices in TTC staining solution for about 10 to 20 minutes in the dark, then take them out and photograph them under a stereomicroscope.

[0090] (8) Statistical quantitative analysis was performed using Imagin J software.

[0091] raw material:

[0092] AAV-GP-12N vector was purchased from Jima Gene;

[0093] Troponin I (c-Tnl), interleukin-1β (1L-1β) and interleukin-6 (1L-6) kits were purchased from ELISA.

[0094] Example 1: Effects of the RNF130 gene on cells

[0095] 1. Construction of RNF130 overexpression plasmid:

[0096] According to the mRNA of RNF130 gene (NM_021540.4), cDNA was obtained by reverse transcription, and the target gene fragment was obtained by PCR amplification; the target gene fragment was inserted between EcoR I and BamH I of plasmid AAV-GP-12N to construct the overexpression plasmid AAV-GP-12N-RNF130; the empty plasmid AAV-GP-12N without the RNF130 gene inserted was used as a control.

[0097] 2. Construction of RNF130 knockdown siRNA

[0098] RNF130-siRNA-1 (SEQ ID NO.2): GGAUCCCUGGCUCAGUGAATT;

[0099] RNF130-siRNA-1 (SEQ ID NO.3): UUCACUGAGCCAGGGAUCCTT;

[0100] NC-siRNA-1 (SEQ ID NO.4):UCUCCGAACGUGUCACGUTT;

[0101] NC-siRNA-2 (SEQ ID NO. 5): ACGUGACACGUUCGGAGAATT.

[0102] 3. Detection of the effect of RNF130 gene on mouse heart cells

[0103] Mouse cardiac cells were isolated and the effects of overexpression or knockout of RNF130 on apoptosis of mouse cardiac cells were detected. The cells were divided into control group (CON), hypoxia / reoxygenation + empty vector group (HR+Vector), hypoxia / reoxygenation + RNF130 overexpression group (HR+RNF130 OE), hypoxia / reoxygenation + NC group (HR+siRNA NC), and hypoxia / reoxygenation + RNF130 knockout group (HR+RNF130 KO), as shown in Table 1.

[0104] Table 1 Cell experiment groups

[0105]

[0106] Cell apoptosis was detected. Figure 1 , Figure 2 As shown, the results showed that overexpression of RNF130 could significantly reduce cell apoptosis, while knockout of RNF130 further aggravated cell apoptosis.

[0107] Example 2: RNF130 activator for the treatment of cardiac ischemia-reperfusion

[0108] 1. Virus construction

[0109] The overexpression plasmid AAV-GP-12N-RNF130 prepared in Example 1 was commissioned to Genema to construct adeno-associated virus RNF130-AAV9 for animal experiments; the empty plasmid was used as a control.

[0110] 2. Animal experiments

[0111] Male C57BL / 6 mice were purchased from Sibeifu (Beijing) Biotechnology Co., Ltd. All mice were housed in specific pathogen-free microisolator cages with free access to food and water, and the animals were housed in a temperature- and humidity-controlled room with a 12-h light / dark cycle.

[0112] The therapeutic effect of RNF130 activator on myocardial ischemia-reperfusion mice was detected, and the groups were shown in Table 2.

[0113] Table 2 Mouse experimental groups

[0114]

[0115]

[0116] The mouse heart and serum indicators were tested and the results were as follows:

[0117] The results of left ventricular ejection fraction (EF) and left ventricular fractional shortening (FS) in mice were as follows Figure 3 A, 3B, the results of chest echocardiography of mice are as follows Figure 3 D. The results showed that the EF and FS of the MI / IR+Vector group were significantly lower than those of the Sham group, and overexpression of RNF130 could significantly improve the EF and FS indicators of heart-ischemia-reperfusion mice;

[0118] The levels of cTnl (troponin), IL-1β, and IL-6 in the blood Figure 3As shown in C, the results showed that cTnl, IL-1β, and IL-6 in the MI / IR+Vector group were significantly higher than those in the Sham group, indicating that the MI / IR+Vector group had myocardial damage and inflammatory response, and overexpression of RNF130 could significantly improve myocardial damage and reduce inflammation.

[0119] The expression results of caspase-3 protein in mouse heart are as follows Figure 4 As shown in A, the results showed that the expression of caspase-3 protein was high in the MI / IR+Vector group, indicating severe myocardial damage; the expression of caspase-3 protein was significantly decreased in the MI / RI+RNF130 OE group;

[0120] Mouse heart DHE staining results are as follows Figure 4 As shown in B, the results showed that the accumulation of ROS in the hearts of mice in the MI / IR+Vector group was high and the oxidative stress was severe; the accumulation of ROS in the MI / RI+RNF130 OE group was significantly reduced;

[0121] The fluorescence detection results of TUNEL are as follows Figure 4 As shown in C, the results showed that the TUNEL fluorescence intensity in the hearts of mice in the MI / IR+Vector group increased, indicating severe cell apoptosis. The TUNEL fluorescence intensity in the MI / RI+RNF130 OE group was significantly lower than that in the MI / IR+Vector group, indicating that overexpression of RNF130 can significantly reduce cell apoptosis caused by cardiac ischemia-reperfusion.

[0122] Evans blue & TTC staining results of mouse heart Figure 5 A, The results showed that the size of myocardial infarction in mice was detected, and it was observed that the infarct area in RNF130OE mice was significantly reduced compared with that in the MI / IR+Vector group mice;

[0123] The extent of myocardial infarction in mice Figure 5 As shown in B, the results showed that the myocardial infarction area of ​​mice in the MI / IR+Vector group was significantly increased compared with that in the Sham group; the myocardial infarction area of ​​mice in the MI / RI+RNF130 OE group was significantly decreased compared with that in the MI / IR+Vector group.

[0124] The fluorescence intensity results of F4 / 80 (macrophage-specific marker) in mouse heart are as follows Figure 6 As shown, the results showed that macrophages accumulated in the hearts of mice in the MI / IR+Vector group and the degree of inflammation was high; the F4 / 80 fluorescence intensity in the MI / RI+RNF130 OE group was significantly lower than that in the MI / IR+Vector group.

[0125] RNF130 cDNA gene sequence (SEQ ID NO.1):

[0126]

[0127] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. Application of RNF130 as a therapeutic target in the preparation of drugs for preventing or treating ischemic heart disease.

2. The use according to claim 1, characterized in that: The drug increases the expression of the RNF130 gene or protein.

3. The use according to claim 1, characterized in that: The drug contains an activator of RNF130.

4. The use according to claim 3, characterized in that: The activator includes an agent that enhances the expression of the RNF130 gene and / or an agent that enhances the expression of the RNF130 protein.

5. The use according to claim 1, characterized in that: The drug improves myocardial damage and / or cardiac tissue pathological changes induced by cardiac ischemia-reperfusion injury; reduces the expression levels of myocardial infarction markers c-Tnl, 1L-1β and 1L-6; and inhibits the expression of inflammation and oxidative damage markers NOX-2, BAX, and TNF-α proteins.

6. Use of an RNF130 activator in the preparation of a drug for preventing or treating ischemic heart disease, characterized in that: The RNF130 activator includes an agent that enhances the expression of the RNF130 gene and / or an agent that enhances the expression of the RNF130 protein.

7. Application of RNF130 gene or protein in screening drugs for preventing or treating ischemic heart disease.

8. A drug capable of preventing or treating ischemic heart disease, characterized in that: The drug comprises an activator of the RNF130 gene and / or its expression product.

9. The drug according to claim 8, characterized in that The drug includes pharmaceutically acceptable excipients.

10. The drug according to claim 8, characterized in that The drug is in a pharmaceutically acceptable dosage form.

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