SLC25A4 gene and interferent and application thereof

Abstract

The invention provides an SLC25A4 gene as well as an interferent and application thereof, and relates to the technical field of biomedicine. The SLC25A4 gene interferent is injected into a mouse body and is continuously released, so that the processes of mitochondrial injury and pyroptosis are inhibited, and the myocardial fibrosis and cardiac remodeling after myocardial infarction are reduced.

Description

Technical Field

[0001] The present invention belongs to the field of medical biotechnology, and relates to a new SLC25A4 gene signature and its application, in particular to a treatment for preventing and treating myocardial mitochondrial damage and myocardial pyroptosis with an SLC25A4 inhibitor and its application. Background Art

[0002] Disclosing the information of this background art section is only intended to enhance the understanding of the overall background of the present invention, and is not necessarily to be regarded as an admission or an implication in any form that this information constitutes the prior art already known to those of ordinary skill in the art.

[0003] Ischemic heart disease is the main cause of death from cardiovascular diseases worldwide, and myocardial infarction is the most prominent form among them, mainly manifested as irreversible loss of myocardial cell function, and ultimately developing into heart failure, seriously affecting the quality of life of patients. According to the report released by the World Health Organization, heart failure is a global disease that affects approximately 38 million people worldwide, having a huge negative impact on the quality of human life. Only in the United States, the annual financial burden exceeds $30.7 billion, and it is expected to increase to nearly $70 billion by 2030. There are approximately 2 to 4 billion myocardial cells in the human left ventricle, and about 0.5 to 1 billion myocardial cells will be lost within a few hours of a myocardial infarction. The treatment focus of ischemic heart disease is to protect the heart from further developing into heart failure. Currently, reperfusion through thrombolysis, cardiac intervention, and bypass surgery can improve blood supply and rescue injured ischemic myocardial tissue. However, the existing treatment methods essentially cannot improve the irreversible loss of myocardial cells in patients, nor are there ideal drugs to fundamentally reverse the prognosis of heart function. Therefore, in order to alleviate the pain caused by ischemic heart disease clinically, it is urgent to study a new drug for treating and improving the prognosis of patients' heart function.

[0004] In the pathophysiological process of the cardiovascular system, the translocation of intestinal bacteria and their metabolites leads to impaired intestinal barrier function and circulating inflammatory responses, further exacerbating myocardial injury. Previous studies have shown that the circulating level of trimethylamine N-oxide (TMAO) is significantly increased in patients with myocardial infarction. TMAO is a gut metabolite, and an increase in its plasma level can trigger the pathological processes of cellular reactive oxygen species, inflammatory signaling pathways, and various metabolic diseases. Ultrastructurally, TMAO-treated myocardial cells show glycogen accumulation, increased mitochondrial fission, and lipofuscin deposition, indicating altered cellular energy metabolism and increased oxidative damage. The latest research shows that the inner mitochondrial membrane protein SLC25A4 is involved in regulating mitochondrial membrane permeability transition pore (MPTP)-dependent cell death and muscular dystrophy pathophysiological processes. Deletion of the SLC25A4 gene promotes the activation and desensitization of MPTP in mitochondria, preventing the progression of cell death.

[0005] Recent studies have shown that myocardial injury can soon trigger pyroptosis of cardiomyocytes and immune-inflammatory regulation. However, how TMAO regulates SLC25A4-induced pyroptosis of cardiomyocytes and pathological cardiac remodeling after myocardial injury remains unclear.

[0006] The paper "Single-cell transcriptomics in MI identify SLC25A4 as a new modulator of mitochondrial malfunction and apoptosis-associated cardiomyocyte subcluster" discloses that the expression of SLC25A4 increases in failing hearts, and the downregulation of SLC25A4 improves mitochondrial function and reduces apoptosis. However, the role of SLC25A4 in the regulation of pyroptosis of cardiomyocytes has not been involved. Summary of the Invention

[0007] In view of the current diagnosis and treatment status of the poor clinical prognosis of ischemic myocardial infarction, the main object of the present invention is to provide a new intervention target that can effectively reduce myocardial mitochondrial disorders and heart damage induced by circulating TMAO. By clarifying the process of mitochondrial SLC25A4 regulating the phenotypes of myocardial pyroptosis and cardiac mitochondrial function, the present invention reveals potential intervention targets and inflammatory signaling pathways for myocardial pyroptosis and repair after myocardial injury, thereby enhancing the potential for clinical translation and the level of diagnosis and treatment of myocardial repair, and providing important research value and clinical significance for the treatment of cardiovascular diseases.

[0008] To achieve the above object, the present invention adopts the following technical solutions:

[0009] In the first aspect of the present invention, there is provided an application of a detection reagent for SLC25A4 gene or the protein encoded thereby in the preparation of a product for the auxiliary diagnosis of TMAO-induced myocardial mitochondrial damage and pyroptosis.

[0010] The present invention discovers that TMAO significantly promotes the increase in the expression level of SLC25A4. Therefore, SLC25A4 may be a key target protein molecule involved in the mechanism of TMAO-induced pyroptosis of cardiomyocytes in cardiovascular diseases. Through the study of SLC25A4 in the heart, interfering substances of SLC25A4 are injected intramuscularly or into the tail vein of mice to achieve sustained release, thereby improving myocardial contractile function, reducing scar formation, and promoting the long-term clinical prognosis of cardiac function in patients with myocardial infarction.

[0011] In some embodiments, the amino acid sequence of the protein encoded by SLC25A4 is as shown in SEQ ID NO.1.

[0012] SEQ ID NO.1:

[0013] MGDQALSFLKDFLAGGIAAAVSKTAVAPIERVKLLLQVQHASKQISAEKQYK

[0014] GIIDCVVRIPKEQGFLSFWRGNLANVIRYFPTQALNFAFKDKYKQIFLGGVDR

[0015] HKQFWRYFAGNLASGGAAGATSLCFVYPLDFARTRLAADVGKGSSQREFNG

[0016] LGDCLTKIFKSDGLKGLYQGFSVSVQGIIIYRAAYFGVYDTAKGMLPDPKNVH

[0017] IIVSWMIAQSVTAVAGLVSYPFDTVRRRMMMQSGRKGADIMYTGTLDCWRKIAKDEGANAFFKGAWSNVLRGMGGAFVLVLYDEIKKYV。

[0018] The expression of the SLC25A4 gene provided by the present invention increases with the increase of TMAO level and is up-regulated in myocardial infarction tissues, suggesting that SLC25A4 can be used as a new target for heart repair and has important research value in the treatment of cardiovascular diseases. Therefore, intramuscular or tail vein injection of SLC25A4 interfering substances into mice can effectively alleviate myocardial injury, thereby achieving the purpose of preventing or treating heart diseases.

[0019] In a second aspect of the present invention, there is provided the use of the SLC25A4 gene or the encoded protein as a biomarker in the preparation of a drug for preventing and / or treating TMAO-induced myocardial mitochondrial damage, and the amino acid sequence of the protein encoded by SLC25A4 is as shown in SEQ ID NO.1.

[0020] In some embodiments, the drug comprises an SLC25A4 inhibitor, and the SLC25A4 inhibitor comprises: an adeno-associated virus that knocks down SLC25A4 or siRNA encapsulated in liposomes.

[0021] In a third aspect of the present invention, there is provided the use of an interfering substance that inhibits the expression and / or function of SLC25A4 in the preparation of a drug for preventing, alleviating or / and treating myocardial ischemia injury.

[0022] In some embodiments, the drug comprises: small interfering nucleic acid siSLC25A4 of SLC25A4, and the nucleic acid sequence of the siSLC25A4 is as shown in SEQ ID NO.2. After knocking down SLC25A4 in cardiomyocytes, the expression levels of mitochondrial ROS, mtDNA and cardiomyocyte pyroptosis protein GSDMD can be inhibited.

[0023] SEQ ID NO.2:

[0024] CAGGCUUUGAGCUUUCUUATT UAAGAAAGCUCAAAGCCUGTT.

[0025] In a fourth aspect of the present invention, there is provided the use of an interfering agent that inhibits the expression and / or function of SLC25A4 in the preparation of a drug for preventing, alleviating or / and treating heart diseases. The SLC25A4 gene interfering agent enters the body through intramuscular injection and tail vein in mice and exerts its drug effect, and is a cardiovascular drug for preventing and / or treating heart diseases.

[0026] In some embodiments, the drug comprises: an SLC25A4 gene interfering agent, and the SLC25A4 gene interfering agent is an AAV-9 adeno-associated virus inhibitory vector, and its nucleic acid sequence is as shown in SEQ ID NO.3. Intravenous injection into mice can significantly inhibit the protein expression of SLC25A4. The research results show that inhibiting SLC25A4 significantly regulates key events such as TMAO-induced myocardial mitochondrial ROS, membrane potential and cardiomyocyte pyroptosis, promotes cardiomyocyte repair, and thus improves the clinical prognosis of myocardial infarction.

[0027] SEQ ID NO.3:

[0028] CAGGGCATCATCATCTACA.

[0029] In some embodiments, the heart diseases include cardiac remodeling, myocardial fibrosis, coronary heart disease, myocardial infarction or heart failure.

[0030] Advantages of the present invention

[0031] (1) The present invention uses the SLC25A4 gene as a marker for TMAO-induced myocardial mitochondrial damage and cardiomyocyte pyroptosis. By preparing its interfering agent and injecting it into mice. The SLC25A4 interfering agent provided by the present invention can be used as an intervention drug for the repair after myocardial injury. Inhibiting the expression of SLC25A4 can significantly reduce the level of myocardial mitochondrial damage, inhibit cardiomyocyte pyroptosis and then activate the repair of damaged tissues. The SLC25A4 interfering agent of the present invention effectively alleviates the problems of poor prognosis of myocardial infarction patients and the treatment of heart diseases.

[0032] (2) Different from the existing research which believes that the down-regulation of SLC25A4 reduces apoptosis, the present invention for the first time discovers that during the occurrence of myocardial infarction injury, TMAO promotes the protein level expression activity of SLC25A4 and is closely related to cardiomyocyte pyroptosis; silencing SLC25A4 can significantly inhibit mitochondrial damage, cardiomyocyte pyroptosis and the area of myocardial infarction. Therefore, SLC25A4 serves as a key intervention target of TMAO in the regulation of cardiomyocyte pyroptosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] The accompanying drawings forming a part of this invention are used to provide a further understanding of the invention. The exemplary embodiments of the invention and their descriptions are used to explain the invention and do not constitute an improper limitation of the invention.

[0034] Figure 1 To detect the expression level of plasma TMAO after mice were fed a high-choline diet for 3 weeks.

[0035] Figure 2 After treating cardiomyocytes with TMAO, the protein expression level of SLC25A4 was detected by Western blot.

[0036] Figure 3 After transfecting cardiomyocytes with siSLC25A4 and siNC, the expression of SLC25A4 in cardiomyocytes was detected by immunofluorescence.

[0037] Figure 4 Under hypoxic conditions of cardiomyocytes, after knocking down SLC25A4 in TMAO-treated cardiomyocytes, the effect on the level of mitochondrial superoxide in cardiomyocytes was detected by immunofluorescence.

[0038] Figure 5 Under hypoxic conditions of cardiomyocytes, after knocking down SLC25A4 in TMAO-treated cardiomyocytes, the effect on the translocation of mtDNA in cardiomyocytes was detected by immunofluorescence.

[0039] Figure 6 Under hypoxic conditions of cardiomyocytes, after knocking down SLC25A4 in TMAO-treated cardiomyocytes, the effect on the expression of the pyroptosis protein GSDMD in cardiomyocytes was detected by Western blot.

[0040] Figure 7 After injecting adeno-associated virus shSLC25A4 into mice, the effect on the cardiac ejection fraction after myocardial infarction in high-choline diet mice was detected to evaluate the therapeutic effect of the SLC25A4 interfering agent.

[0041] Figure 8After injecting adeno-associated virus of shSLC25A4 into mice, the effect of high-choline diet on the level of cardiac fibrosis after myocardial infarction in mice was detected to evaluate the therapeutic effect of SLC25A4 interference substances. Detailed implementation mode

[0042] It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those of ordinary skill in the technical field to which the present invention belongs.

[0043] The present invention will be further described in detail below in conjunction with specific embodiments. It should be noted that the specific embodiments are interpretations rather than limitations of the present invention.

[0044] Example 1 Quantitative analysis and detection of TMAO in mouse plasma

[0045] In the present invention, male C57BL6 / J mice aged 6-8 weeks were used. 1.2% choline was added to the standard rodent feed, and the expression level of plasma TMAO was detected after 3 weeks of high-choline diet in mice. Mouse plasma samples (n = 5) were thawed at 4°C, and isotope internal standard and pre-cooled acetonitrile / water solution (9 / 1, v / v) were added, and vortexed to extract metabolites. The chromatographic peak area and retention time were extracted by Multiquant 3.0.2 software. The retention time was corrected with the standard substance of the target substance for metabolite identification and quantitative analysis ( Figure 1 ).

[0046] Example 2 Synthesis and effect verification of small interfering RNA (siRNA) of SLC25A4

[0047] (1) According to the SLC25A4 sequence, the siRNA interference sequence (siSLC25A4) of SLC25A4 and its control siRNA NC (siNC) were designed and synthesized by GenePharma. The nucleic acid sequence of siSLC25A4 is shown in SEQ ID NO.2.

[0048] (2) After siSLC25A4 and siNC were transfected into primary cardiomyocytes for 24 hours respectively, the expression of SLC25A4 was detected by immunofluorescence (see Figure 3 ).

[0049] Example 3 Construction and effect evaluation of mitochondrial SLC25A4 adeno-associated virus vector

[0050] (1) According to the SLC25A4 gene sequence, the adeno-associated virus vector was designed and synthesized by Obio Technology. The SLC25A4 interference nucleic acid sequence is shown in SEQ ID NO.3.

[0051] (2) Inject adeno-associated virus of SLC25A4 into mice via the vein.

[0052] (3) Name the adeno-associated virus of SLC25A4 and its control as NC and shSLC25A4 respectively. Two weeks after injecting them into mice, use echocardiogram (see Figure 7 ) and cardiac Masson staining to detect the therapeutic effect of the SLC25A4 interfering agent (see Figure 8 ).

[0053] Example 4 Isolation and culture of neonatal mouse ventricular myocytes

[0054] (1) Experimental preparation. The whole experimental operation process is carried out in a sterile workbench. Prepare 0.2 mg / ml trypsin (P-3292, Sigma) and 1.0 mg / ml collagenase II (LS004176, Worthington) for digestion of cardiac tissue.

[0055] (2) Put 50 neonatal mice aged 1 - 3 days into a sterile container, expose the heart, and use forceps to take out the heart and place it in ice-cold PBS buffer.

[0056] (3) Wash the heart 3 times, use surgical scissors to cut the cardiac tissue into pieces and put them into a conical flask, add the digestive solution, and lyse and digest at 42 °C for 5 min, while constantly shaking the conical flask.

[0057] (4) Transfer the supernatant of the digestive solution to a centrifuge tube containing 5 mL of serum to terminate digestion, centrifuge the collected digestion product at 1000 rpm for 5 min, and collect the precipitate.

[0058] (5) Filter the collected cell suspension into a 10 mL cell culture dish, put it into a cell culture incubator, and perform differential adhesion for 1 - 1.5 h.

[0059] (6) Take the cell suspension, add Brdu to inhibit the proliferation of non-myocardial cells, and aliquot it into six-well plates, then put it into a carbon dioxide incubator for further culture.

[0060] (7) After 24 h, change the neonatal mouse ventricular myocytes to a new DMEM / F12 medium. After treating the myocytes with TMAO (600 μM) for 24 h, collect the cells for Western blots to detect the expression level of SLC25A4.

[0061] Example 5 Extraction of total protein from myocytes and Western blot analysis

[0062] (1) Take an appropriate amount of lysis buffer and add protease inhibitor at a ratio of 1:100 before use.

[0063] (2) Add 1 mL of PBS to each well in a six-well plate, and collect the cells into a centrifuge tube using a cell scraper. Centrifuge at 4°C and 500 g for 5 min. Add the prepared lysis buffer, pipette and mix several times. After thorough mixing, place on ice for 20 min for lysis.

[0064] (3) Centrifuge at 4°C and 12,000 g for 10 min. Pipette and transfer the supernatant to a new centrifuge tube.

[0065] (4) Add protein loading buffer, boil at 100°C for 5 - 10 min, measure the protein concentration by BCA method or store at -20°C in the refrigerator.

[0066] (5) Electrophoresis. Preparation of SDS-PAGE gel: 5% upper gel (stacking gel), 12% lower gel (separating gel). After the SDS-PAGE gel is set, add an appropriate amount of protein solution. The voltage for the sample in the stacking gel is 80 V for 30 min, and after entering the separating gel, the voltage is adjusted to 120 V for 1.5 h.

[0067] (6) Transfer. After electrophoresis in the SDS-PAGE gel, transfer the gel to a nitrocellulose membrane. Control the transfer current at 280 mA for 1.5 h.

[0068] (7) Blocking. Transfer the nitrocellulose membrane to TBST containing 5% non-fat milk powder and incubate at room temperature for 1 h.

[0069] (8) Antibody incubation. Dilute the SLC25A4 or GSDMD antibody with 5% BSA in proportion and incubate at room temperature for 2 h or overnight at 4°C.

[0070] (9) Wash the membrane. Wash the primary antibody with TBST 3 times, 5 min each time.

[0071] (10) Antibody incubation. Dilute the secondary antibody (horseradish peroxidase-labeled secondary antibody) with 2% BSA in proportion and incubate at room temperature for 1 h.

[0072] (11) Wash the membrane. Wash the primary antibody with TBST 3 times, 5 min each time.

[0073] (12) Develop the film. Incubate the membrane with enhanced ECL developing solution for 5 min, and develop and analyze the expression of SLC25A4 and GSDMD proteins using a scanning imager ( Figure 2 and Figure 6 ).

[0074] Example 6 Immunofluorescence Detection of the Expression of Mitochondrial mtDNA in Cardiomyocytes

[0075] (1) Wash the cardiomyocytes transfected with siSLC25A4 and siNC 3 times with PBS, 3 min each time.

[0076] (2) Fix with 4% paraformaldehyde for 15 min, and wash 3 times with PBS, 3 min each time.

[0077] (3) Permeabilize with 0.5% TritonX-100 at room temperature for 15 min.

[0078] (4) Block with 5% BSA for 30 min.

[0079] (5) Discard the blocking solution, add primary antibodies (anti-TFAM, 1:200; anti-cTnT, 1:500), incubate at room temperature for 2 h or overnight at 4°C. Wash 3 times with PBST, 3 min each time.

[0080] (6) Add fluorescent secondary antibody, incubate at room temperature in the dark for 1 h. Wash 3 times with PBST, 3 min each time.

[0081] (7) Stain with DAPI for 5 min. Wash 3 times with PBST, 3 min each time.

[0082] (8) Add anti-quenching mounting medium and cover with a coverslip.

[0083] (9) Observe and take pictures under a fluorescence microscope or a laser confocal microscope. By counting the number of positive cells of myocardial cell mtDNA and ROS, the membrane potential of cardiomyocytes transfected with siSLC25A4 was restored, and the translocation of mtDNA was reduced ( Figure 5 ).

[0084] Example 7 Detection of mitochondrial superoxide (MitoSOX) in cardiomyocytes

[0085] After transfection of cardiomyocytes with siSLC25A4 and siNC, staining was performed using the MitoSOX Red (S0061S) detection kit. Briefly, for one well of a 6-well plate, aspirate the culture medium and wash the cells 3 times with PBS solution, 3 min each time. Subsequently, add 1 mL of MitoSOX Red staining working solution and incubate at 37°C for 10 - 30 min. Aspirate the supernatant and wash 3 times with PBS, and observe under a fluorescence microscope or a laser confocal microscope ( Figure 4 ).

[0086] Example 8 Application of SLC25A4 gene silencer in cardiac repair after myocardial infarction;

[0087] In this study, 6 - 8-week-old male C57BL6 / J mice were used. 1.2% choline was added to the standard rodent diet, and the mice were fed a high-choline diet for 3 weeks and then underwent MI surgery. Left ventricular function and structure were evaluated by two-dimensional echocardiography. The specific implementation is as follows:

[0088] (1) Construction of mouse myocardial infarction model and administration

[0089] An adult male mouse myocardial infarction model was constructed by permanent ligation of the left anterior descending coronary artery. The mouse was placed on a 37 °C constant temperature heating pad in a left lateral decubitus position under general anesthesia. The animal was intubated tracheally, and the parameters of the animal ventilator were adjusted to maintain normal respiration in the mouse. The adult mouse was thoracotomized between the 3rd and 4th intercostal spaces, the chest cavity was opened with a rib retractor, the heart was exposed, the pericardium was carefully removed, and the left anterior descending coronary artery was identified. The left anterior descending coronary artery (LAD) was ligated with a polypropylene suture (6-0) 2-3 mm distal to the ascending aorta, and the ligation induced myocardial infarction. When the left ventricle became pale, the ligation was considered successful. Then, the ribs were closed and the skin was sutured with a 4-0 polypropylene suture, and the mouse was placed in a warm and comfortable environment and returned to the cage for feeding after complete recovery.

[0090] (2) Echocardiographic analysis of cardiac function in mice

[0091] Echocardiography was performed on high-choline mice 14 days after myocardial infarction; transthoracic echocardiography was performed using a Vevo100 imaging system equipped with a 40 MHz MS-250 scanning head; the ejection fraction (EF) was calculated according to the established standard equation; it was found through analysis of echocardiographic images that the SLC25A4 inhibitor significantly restored the systolic and diastolic functions of the heart after myocardial infarction.

[0092] (3) Masson staining and analysis of heart tissue

[0093] Masson trichrome staining, also known as Masson staining, is an authoritative and classic technical method for collagen fiber staining. After Masson staining, muscle fibers are red and collagen fibers are green or blue, mainly used to distinguish collagen fibers and muscle fibers.

[0094] First, the hearts of mice 14 days after infarction were fixed, dehydrated, embedded, sectioned and dewaxed to water; Weigert iron hematoxylin staining solution was prepared with Weigert stain A and Weigert stain B and stained at room temperature for 6 min; differentiated with acidic ethanol differentiating solution for 10 s and washed with double-distilled water 2-3 times; several drops of bluing solution were added to the tissue section and washed with double-distilled water for 3 min; stained with Ponceau fuchsin staining solution for 8 min and washed with 0.2% acetic acid solution for 3 min; washed with phosphomolybdic acid solution for 2 min and washed with 0.2% acetic acid solution for 3 min; stained with aniline blue staining solution for 1 min and washed with 0.2% acetic acid solution for 3 min; dehydrated with gradient ethanol, cleared with xylene twice, sealed with neutral gum, and photographed after natural drying.

[0095] In summary, SLC25A4 is a potential therapeutic target in cardiomyocyte repair, and the application of SLC25A4 inhibitors effectively alleviates TMAO-induced myocardial mitochondrial damage and the development of myocardial pyroptosis. Stable inhibition of SLC25A4 gene expression in vivo can significantly reduce the infarct size and improve cardiac function, revealing the important application value of SLC25A4 inhibitors in the treatment of myocardial infarction.

[0096] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and changes. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

Use of a detection reagent for the SLC25A4 gene or the protein encoded thereby in the preparation of a product for the auxiliary diagnosis of TMAO-induced myocardial mitochondrial damage.

2. The application according to claim 1, characterized in that, The amino acid sequence of the protein encoded by SLC25A4 is as shown in SEQ ID NO.

1.

3. Use of the SLC25A4 gene or the protein encoded thereby as a biomarker in the preparation of a drug for preventing and / or treating TMAO-induced myocardial mitochondrial damage, characterized in that, The amino acid sequence of the protein encoded by SLC25A4 is as shown in SEQ ID NO.

1.

4. The application according to claim 3, characterized in that The drug comprises an SLC25A4 inhibitor, and the SLC25A4 inhibitor comprises: an adeno-associated virus that knocks down SLC25A4 or siRNA encapsulated in liposomes.

5. Use of an interfering agent that inhibits the expression and / or function of SLC25A4 in the preparation of a drug for preventing, alleviating or / and treating myocardial ischemia injury.

6. The application according to claim 5, characterized in that, The drug comprises: small interfering nucleic acid siSLC25A4 of SLC25A4, and the nucleic acid sequence of siSLC25A4 is as shown in SEQ ID NO.

2.

7. The application according to claim 5, wherein The drug can inhibit the expression levels of mitochondrial ROS, mtDNA and the cardiomyocyte pyroptosis protein GSDMD.

8. Use of an interfering agent that inhibits the expression and / or function of SLC25A4 in the preparation of a drug for preventing, alleviating or / and treating heart diseases.

9. The application according to claim 8, characterized in that, The drug comprises: an SLC25A4 gene interfering agent, and the SLC25A4 gene interfering agent is an AAV-9 adeno-associated virus inhibitory vector, and its nucleic acid sequence is as shown in SEQ ID NO.

3.

10. The application according to claim 8, wherein, The heart diseases include cardiac remodeling, myocardial fibrosis, coronary heart disease, myocardial infarction or heart failure.

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