Use of slco5a1 in the preparation of a medicament for the treatment and / or prevention of heart failure

Abstract

The application relates to application of SLCO5A1 in preparation of a medicine for treating and / or preventing heart failure, the medicine up-regulating expression of SLCO5A1 protein in myocardial cells; and the active ingredient of the medicine comprises 3-iodothyronine. The application first proposes application of the SLCO5A1 target point in treatment of heart failure, discloses that SLCO5A1 is differentially expressed in multiple heart failure animal models and the pharmacological effect molecule 3-iodothyronine, and provides a novel treatment strategy of an action mechanism for heart failure treatment.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, and in particular to the use of SLCO5A1 in the preparation of drugs for the treatment and / or prevention of heart failure. Background Technology

[0002] Heart failure (HF) is the end stage of various cardiovascular diseases, characterized by high mortality and disability rates. Currently, the incidence of HF is rising annually, imposing a heavy economic burden on individuals and society, yet effective treatments remain lacking.

[0003] Currently, the clinical treatment strategy for heart failure has shifted from the traditional "cardiac strengthening, diuresis, and vasodilation" model to a comprehensive treatment model that improves hemodynamics while blocking excessive activation of the neuroendocrine system and inhibiting myocardial remodeling. Commonly used drugs include angiotensin-converting enzyme inhibitors (ACEIs), angiotensin II receptor antagonists (ARBs), beta-blockers, aldosterone receptor antagonists (MRAs), and sodium-glucose cotransporter 2 inhibitors (SGLT2i). Although these drugs can significantly improve patient prognosis, they cannot completely control disease progression, and their use is limited in patients with hemodynamic abnormalities, potentially leading to reduced tolerability and compliance. However, the field of heart failure drug therapy still faces significant challenges. Many drugs, while relieving symptoms, are unlikely to reverse myocardial remodeling or significantly reduce mortality in advanced-stage patients. Long-term use of these drugs may cause side effects such as electrolyte disturbances, bradycardia, and hypotension. Furthermore, existing drugs primarily target the hemodynamic and neurohormonal systems, lacking specificity for other molecular targets in heart failure, thus limiting their efficacy. Therefore, discovering therapeutic drugs with novel targets that can effectively improve prognosis is a scientific problem that urgently needs to be solved in clinical practice.

[0004] SLCO5A1 (5A1) is a member of the solute carrier organic anion transporter family. It mediates the transmembrane transport of various endogenous and exogenous organic anions, including bile acids, prostaglandins, thyroxines, and various drugs. Currently, there are no reports on its function in cardiovascular diseases such as myocardial infarction and heart failure. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides the use of SLCO5A1 in the preparation of drugs for the treatment and / or prevention of heart failure, characterized in that the drugs upregulate the expression of SLCO5A1 protein.

[0006] In one embodiment, the drug upregulates the expression of SLCO5A1 protein in cardiomyocytes.

[0007] In one embodiment, the active ingredient of the drug includes 3-iodothyronine (T1AM).

[0008] In one embodiment, the drug further includes pharmaceutically acceptable excipients.

[0009] In one embodiment, the excipients include at least one of excipients, lubricants, antioxidants, preservatives, binders, fillers, or thickeners.

[0010] In one embodiment, the dosage of the drug is 10-50 mg / kg.

[0011] A second aspect of the invention also provides the use of SLCO5A1 as a target in screening drugs for the treatment and / or prevention of heart failure.

[0012] A third aspect of the invention also provides the use of 3-iodothyronine in the preparation of medicaments for the treatment and / or prevention of heart failure.

[0013] In one embodiment, the drug further includes pharmaceutically acceptable excipients.

[0014] A fourth aspect of the present invention also provides a method for screening drugs for the treatment and / or prevention of heart failure, characterized in that, when the test component upregulates the expression of SLCO5A1 protein or enhances SLCO5A1 activity, it is determined that the test component is highly likely to treat and / or prevent heart failure, and the test component is used as a candidate drug for subsequent testing.

[0015] In addition, the present invention also provides a medicament for treating and / or preventing heart failure, wherein the active ingredient of the medicament includes 3-iodothyronine.

[0016] This invention is the first to propose the application of the SLCO5A1 target in the treatment of heart failure. Compared with existing technologies, this invention has the following advantages: 1. SLCO5A1 is independent of the neuroendocrine system, avoiding the limitations of electrolyte imbalance and neurological dysfunction.

[0017] 2. SLCO5A1 is a key functional molecule regulating heart failure by modulating the transport of endogenous substances. This invention reveals for the first time the differential expression of SLCO5A1 in multiple animal models of heart failure and demonstrates the effectiveness of intervention based on this target in treating heart failure.

[0018] 3. The pharmacological effector molecule of SLCO5A1 was found to be 3-iodothyronine (T1AM), proving its application value. Attached Figure Description

[0019] Figure 1 This image shows the downregulation of SLCO5A1 protein expression in myocardial tissue and hypertrophic cardiomyocytes in heart failure. AB represents the results of Western blot analysis and statistical analysis of SLCO5A1 protein expression in myocardial tissue of mice 28 days after myocardial infarction (HF) and in the sham-operated group. CD represents the results of Western blot analysis of SLCO5A1 protein expression in myocardial tissue of TAC mice and the sham-operated group 14 days after surgery. EF represents the immunofluorescence assay of SLCO5A1 expression in myocardial tissue of HF mice, using cardiac troponin T (cTNT) co-localization labeling of cardiomyocytes. GH represents the Western blot analysis of SLCO5A1 protein expression in ISO+PE-induced hypertrophic cardiomyocytes. N=6, *versusSham or Ctrl, P<0.05. English captions in the image: Sham → sham surgery; HF → heart failure; TuB → α-tubulin; TAC → aortic arch stenosis model; ISO → isoproterenol; PE → phenylephrine; Ctrl → control group. Figure 2 The diagram shows the upregulation of SLCO5A1 protein expression by T1AM; where A is the chemical structure of T1AM; BC represents the expression of SLCO5A1 protein detected by Western blot after treatment with 1.5 μM (L), 3 μM (M), and 6 μM (H) T1AM for 36 hours in a cardiomyocyte hypertrophy model; DE represents the expression of SLCO5A1 protein in HF mice after administration of 12.5 mg / kg T1AM. -1 (L), 25mg·kg -1 (M), 50 mg·kg -1 (H) After 28 days of treatment, Western blot analysis was performed to detect SLCO5A1 protein expression; *versus NC or Sham, #versus ISO+PE+N or HF-N, P<0.05, n=6; English annotations in the figure: Sham → sham surgery; HF → heart failure; TuB → α-tubulin; NC → negative control; N → solvent control; L → low dose; M → medium dose; H → high dose; Figure 3This figure shows the progression of heart failure in mice inhibited by T1AM; where A is the mouse survival curve, analyzed using the Kaplan-Meier method, N=30; BC shows the short-axis M-mode echocardiogram results and the comparison of left ventricular ejection fraction (EF) and fractional shortening (FS) between groups; DE shows the expression of fibrosis-related proteins detected by Western blot; FG shows the expression of myocardial remodeling marker proteins detected by Western blot; HI shows the detection of cardiomyocyte hypertrophy by WGA (wheat germ lectin) staining; JK shows the detection of myocardial fibrosis by Masson staining; *versus Sham, #versus HF-N, P<0.05, n=6; English labels in the figure: Sham → sham surgery; HF → heart failure; TuB → α-tubulin; N → solvent control group; L → low dose; M → medium dose; H → high dose; MHC → β-myosin heavy chain; ANP → atrial natriuretic peptide; BNP → brain natriuretic peptide; FN → fibronectin; Collagen I / Col → Collagen I; α-SMA / SMA → α-Smooth muscle agonist; Figure 4 The diagram shows how T1AM inhibits myocardial hypertrophy by regulating SLCO5A1. In the diagram, AB represents the expression of SLCO5A1, atrial natriuretic peptide (ANP), and brain natriuretic peptide (BNP) proteins after 36 hours of treatment with 1.5 μM (L), 3 μM (M), and 6 μM (H) T1AM in a myocardial hypertrophy model, as detected by Western blot. CD represents the overexpression of SLCO5A1 in the myocardial hypertrophy model. After 36 hours, Western blot analysis was performed to detect the expression of SLCO5A1, atrial natriuretic peptide (ANP), and brain natriuretic peptide (BNP) proteins. EF represented the treatment of SLCO5A1 expression in a cardiomyocyte hypertrophy model followed by 6 μM (H) T1AM for 36 hours, with Western blot analysis of SLCO5A1, ANP, and BNP protein expression. GH represented the treatment of SLCO5A1 expression in a cardiomyocyte hypertrophy model followed by 6 μM (H) T1AM for 36 hours, with immunofluorescence detection of α-actinin and calculation of cell area. *versus NC or Ctrl, #versus ISO+PE+N, $versus ISO+PE+T1AM (H), P<0.05, n=6; English annotations in the figure: TuB→α-tubulin; NC→negative control; N→solvent control; Ctrl→control group; ANP→atrial natriuretic peptide; BNP→brain natriuretic peptide; L→low dose; M→medium dose; H→high dose; Vector / Vec→vector control; Sh→silence; OE→overexpression; Figure 5Figure 1 shows the effect of SLCO5A1 overexpression on heart failure. AB represents the echocardiogram results of HF mice at 28 days, analyzing left ventricular ejection fraction (EF) and fractional shortening (FS). CF represents Western blot analysis of myocardial remodeling markers (β-MHC, ANP, BNP) and fibrosis markers (Fibronectin, FN, Collagen I, Col; α-SMA) protein expression in HF mice at 28 days. GJ represents Masson staining (G, I) and Picrosirius Red staining (H, J). *versus Sham, #versusHF, P<0.05, n=6. English labels in the figures: Sham → sham surgery; HF → heart failure; OE → overexpression; Vec → vector control; Con → solvent control; TuB → α-tubulin; MHC → β-myocardial myosin heavy chain; ANP → atrial natriuretic peptide; BNP → brain natriuretic peptide; FN → fibronectin; Collagen I → α-sMA. I / Col → Collagen I; α-SMA / SMA → α-smooth muscle agonist. Detailed Implementation

[0020] Current heart failure treatments mostly focus on classic pathways such as regulating cardiomyocyte calcium homeostasis or the adrenergic signaling system. This invention reveals for the first time the crucial role of SLCO5A1, a gene different from traditional pathways, in heart failure, aiming to provide a novel therapeutic strategy with a new mechanism of action, offering an alternative or better option for patients who do not respond well to existing targeted drug therapies.

[0021] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.

[0022] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0023] Unless otherwise specified, all reagents, materials, and equipment used in this embodiment are commercially available; unless otherwise specified, all test methods are conventional test methods in this field.

[0024] Example I. Experimental instruments, reagents, animal cells, etc. 1. Experimental reagents T1AM was purchased from Sigma-Aldrich, USA, with a purity of 98%; 4% paraformaldehyde was purchased from Zhengzhou Pani Chemical Reagent Factory; Masson's trichrome staining kit and Sirius red staining kit were purchased from Solarbio Biotechnology Co., Ltd., China; WGA staining kit was purchased from Sigma-Aldrich, USA; isoflurane for small animals was purchased from Shenzhen Ruiwode Co., Ltd.; physiological saline was purchased from Chenxin Pharmaceutical Co., Ltd.; DMSO and type 2 collagenase were purchased from Sigma-Aldrich, USA; fetal bovine serum was purchased from Zeta Life, USA; 0.25% trypsin, penicillin, and streptomycin were purchased from Shanghai Beyotime Biotechnology Co., Ltd.; PBS and DMEM culture medium were purchased from Wuhan Pronosei Life Sciences Co., Ltd.; SLCO5A1 primary antibody was purchased from Wuhan Sanying Biotechnology Co., Ltd., China.

[0025] 2. Instruments and consumables High-resolution small animal in vivo imaging system (Visual Sonics, Canada); cell culture incubator (Thermo Scientific, USA); electronic balance (Eppendorf, Germany); centrifuge (Thermo Scientific, USA); inverted microscope (Nikon, Japan); optical microscope (Olympus, Japan); confocal microscope (Olympus, Japan); clean bench (Suzhou Purification Co., Ltd.); 6-well and 96-well plates (Wuxi NiceBiotechnology Co., Ltd.); 5ml and 15ml pipettes (Beijing Lanjieke Technology Co., Ltd.); manual pipette (Eppendorf, Germany); 10cm cell culture dish (Wuxi NiceBiotechnology Co., Ltd.).

[0026] 3. Acquisition and culture of primary cardiomyocytes and cardiomyocyte fibroblasts Hearts of 1-day-old C57BL / 6 suckling rats were used. Under aseptic conditions, the hearts were cut into small pieces and digested with 0.125% trypsin and 0.5 mg / mL type 2 collagenase. The resulting cells were cultured in culture dishes for 90 minutes, and cardiomyocytes were obtained by differential adhesion. Primary cardiomyocytes were then obtained by culture in the cell supernatant for 24 hours. The cardiomyocytes were cultured in 0.1% gelatin-coated culture dishes in DMEM containing 10% FBS at a constant temperature of 37°C and 5% CO2 for later use.

[0027] 4. Laboratory animals In this invention, male C57BL / 6 mice were purchased from Guangzhou Tengke Biomedical Technology Co., Ltd., SPF grade, 6-8 weeks old, weighing 20-22g; mouse feed was purchased from Beijing Huafu Biotechnology Co., Ltd.; the experimental animals were housed at the Experimental Animal Center of Guangdong Medical University under constant temperature (25-27℃), constant humidity (45%-50%), fresh air, and dust-free and sterile conditions free from special pathogens (SPE grade). Aseptic feed was provided for the animals to ingest freely, and the bedding was changed every three days. Cages and drinking water were sterilized every three days, and the animals were given sterile distilled water. Aseptic principles were strictly followed when changing feeding supplies. The experimental animals were housed under a 12:12 hour light / dark cycle and had free access to food and water. The animal experiments were approved by the ethics committee of Guangdong Medical University (approval number: GDMU-2025-000166).

[0028] 5. Preparation of pharmaceuticals For cell experiments, the drugs were dissolved in DMSO to 100 μM and then diluted with high-glucose DMEM to the corresponding concentration. For animal experiments, the drugs were dissolved in DMSO to 10 mg / mL, diluted with physiological saline, and then injected intraperitoneally.

[0029] II. Experimental Methods 1. Establishment of cell models and drug treatment (1) Obtaining and culturing primary cardiomyocytes and cardiomyocyte fibroblasts from suckling rats One-day-old hearts of newborn mice were harvested, and the atria and major blood vessels were removed, preserving the ventricular tissue, which was then cut into small fragments. The tissue fragments were digested repeatedly using a combination of 0.125% trypsin and 0.5 mg / mL type II collagenase to release cells. Digestion was terminated with DMEM medium containing 10% serum, and the cells were mixed thoroughly by pipetting and filtered to obtain a mixed cell suspension. This cell suspension was then incubated in culture flasks for 1–1.5 hours, at which point adherent cells developed into cardiomyocytes. The non-adherent cell suspension was collected; this was the preliminarily purified cardiomyocytes, ready for subsequent experiments.

[0030] (2) Preparation of cell hypertrophy model and T1AM treatment Cells were removed from the culture medium and washed twice with phosphate-buffered saline (PBS) and added to a medium containing 1% FBS. Then, 10 μM ISO (isoproterenol) combined with 100 μM PE (phenylephrine) was added for 36 hours to induce a cell hypertrophy model.

[0031] T1AM was pre-incubated with cells at concentrations of 1.5 μM (L), 3 μM (M), and 6 μM (H) for 30 minutes, and then treated with ISO+PE for 36 hours under the same conditions.

[0032] 2. Construction of a mouse heart failure model, intervention, and functional evaluation (1) Mouse heart failure model (HF): 6-8 week old mice were weighed using a small animal balance and anesthetized with 1.5% sevoflurane and 0.6 L / min oxygen. After the anesthesia took effect, the mice were placed in a supine position and fixed on the operating table. Hair removal cream was used to remove hair from the precordial area of ​​the mice, and the surgical area was disinfected with 75% alcohol. The mouse's tongue was lifted with sterile forceps, and then a 1.5 mm endotracheal tube was used to intubate the mouse. After successful intubation, a ventilator was connected for assisted ventilation. The rise and fall of the mouse's chest was observed to ensure that the endotracheal tube was in the trachea, indicating successful intubation. An incision was made between the third and fourth ribs on the left side of the mouse's sternum with sterile scissors. Then, the subcutaneous tissue, pectoralis major muscle, and pectoralis minor muscle were bluntly dissected with sterile forceps, while the intercostal spaces between the third and fourth ribs were bluntly expanded to fully expose the mouse's heart. An 8-0 suture needle was used to ligate the left anterior descending coronary artery by passing through about 1 / 3 of the myocardial layer 2 mm below the lower edge of the left atrial appendage. The muscle was sutured with a purse-string suture for approximately 30 minutes. After 30 minutes, the chest was reopened, and the sutures were cut using sterile suture scissors. At the end of the procedure, the muscle and skin were sutured layer by layer, and the endotracheal tube was removed. The sham-operated group underwent the same procedures as the model group, except that the left anterior descending coronary artery was not ligated. Heart tissue was harvested from mice at the predetermined time point of heart failure (28 days).

[0033] (2) Aortic arch stenosis (TAC): 10-12 week old mice were anesthetized with 1.5% isoflurane and oxygen at a flow rate of 0.6 L / min. The aortic arch was exposed in a supine position. A 26G injection needle was placed at the aortic arch, ligated with 6-0 suture, and then the needle was removed. The thoracic cavity was closed, and the mice were kept warm and revived. Analgesia and antibiotics were administered. The heart was collected 14 days after surgery for testing.

[0034] (3) Intervention: T1AM was administered at doses of 12.5 (L), 25 (M), and 50 (H) mg / kg. -1 Administer intraperitoneal injections once every 3 days until the end of the experiment.

[0035] A cardiomyocyte-specific overexpression vector of SLCO5A1 was constructed using adeno-associated virus vector (AAV9) and cardiac troponin T (cTNT) as promoters, at a concentration of 1×10⁻⁶. 11 Viral genomes are injected via tail vein.

[0036] (4) Ultrasound detection in small animals Mice were induced and maintained using 1.5%–2% isoflurane, and fixed in a supine position on an insulated plate with electrodes for electrocardiogram monitoring. Hair was removed from the mouse's anterior chest area using depilatory cream, and ultrasound coupling agent was applied. Using a mouse ultrasound probe, placed at the left sternal border with the marker pointing towards the right shoulder, the image was slowly adjusted to display a long-axis view from the left atrium to the apex of the heart. After acquiring the B / M image, the probe was rotated 90° clockwise to adjust the display to the level of the papillary muscles, and B-mode ultrasound images were acquired. A sampling line was placed in the center of the left ventricle, and M-mode ultrasound results were acquired, measuring parameters such as left ventricular end-systolic diameter, end-diastolic diameter, ejection fraction, and fractional shortening.

[0037] 3. Semi-quantitative Western blot Cells and myocardial tissues were treated with RIPA lysis buffer containing 1% protease inhibitor, incubated on ice for 30 minutes, and centrifuged at 12,000 rpm at 4°C for 15 minutes. Protein concentration was measured using a BCA protein assay kit. Protein samples were mixed with 5×SDS loading buffer. After boiling at 100°C for 5 minutes, equal volumes of protein were loaded onto 6% or 10% sodium dodecyl sulfate-polyacrylamide gels (SDS-PAGE) for electrophoresis. Separated proteins were transferred to polyvinylidene fluoride (PVDF) membranes, blocked with 5% skim milk at room temperature for 2 hours, incubated overnight with primary antibody at 4°C, and incubated for 2 hours at room temperature with HRP-conjugated secondary antibody. Protein bands were obtained using ECL chemiluminescence reagent.

[0038] 4. Animal tissue pathological staining Fresh heart tissue from mice after modeling was fixed in 4% paraformaldehyde (PFA) for more than 24 hours. After paraffin embedding, the fixed tissue was prepared into 5-micrometer thick sections and subjected to Masson's trichrome staining, Sirius red staining, and WGA staining. Masson's trichrome staining was used to analyze the area of ​​myocardial infarction fibrosis, Sirius red staining was used to assess the degree of collagen deposition in the myocardial interstitium, and WGA staining was used to analyze the cross-sectional area of ​​cardiomyocytes to assess the presence of myocardial hypertrophy. Images of the stained heart tissue sections were acquired using an optical microscope. Masson staining analysis of the degree of myocardial infarction fibrosis was defined as the ratio of the area of ​​left ventricular endocardial scar to the area of ​​the left ventricle; the degree of myocardial interstitial collagen deposition was assessed by calculating the percentage of Sirius red-positive areas to the total field of view using ImageJ software; and WGA staining was performed by measuring the average cross-sectional area of ​​100 cardiomyocytes from 6 randomly selected images in each group using ImageJ software to assess cardiomyocyte size.

[0039] 5. Immunofluorescence staining of animal tissues and cardiomyocytes Fresh heart tissue from mice after modeling was fixed in 4% paraformaldehyde (PFA) for over 24 hours. After paraffin embedding, 5-micrometer thick sections were prepared and dewaxed using gradient ethanol solutions (95%-90%-80%-70%) and pure water. After permeabilization with 0.2% Triton X-100 for 15 minutes and blocking with 5% BSA serum for 1 hour, the sections were incubated overnight at 4°C with SLCO5A1 primary antibody. Following this, the sections were incubated with fluorescently conjugated secondary antibody at room temperature for 2 hours and stained with DAPI (1 μg / mL) for 10 minutes. Images of the stained heart tissue sections were acquired using confocal microscopy. Relative fluorescence intensity was quantified using ImageJ software.

[0040] Adherent cardiomyocytes were fixed with 4% paraformaldehyde (PFA) at room temperature for 30 min after treatment. Following permeabilization with 0.1% Triton X-100 for 5 min and blocking with 5% BSA serum for 1 hour, the cells were incubated overnight at 4°C with α-actinin primary antibody. Subsequently, the cells were incubated with fluorescently conjugated secondary antibody at room temperature for 2 hours and the nuclei were stained with DAPI (1 μg / mL) for 10 min. Fluorescence images of the stained cardiomyocytes were acquired using confocal microscopy. Cell area was quantified relatively using ImageJ software.

[0041] Example 1 Downregulation of SLCO5A1 expression was found in mouse and cellular heart failure models.

[0042] Myocardial tissue was collected on day 28 of myocardial infarction (HF) and day 14 after TAC mouse modeling. Western blotting analysis revealed that SLCO5A1 protein expression in the heart was significantly downregulated. Figure 1 (AD). Furthermore, immunofluorescence assay revealed a significant decrease in SLCO5A1 expression in the cardiac tissue of HF mice after myocardial infarction (AD). Figure 1 The EF (efficacy of SLCO5A1 expression) was consistent with the results of Western blot analysis. A primary neonatal rat cardiomyocyte hypertrophy model was induced using a combination of isoproterenol (ISO) and phenylephrine (PE), and SLCO5A1 expression was also downregulated in this model. Figure 1 (GH). These animal and cellular level results indicate that SLCO5A1 is downregulated during the development and progression of heart failure, and may act as a regulatory molecule for heart failure.

[0043] Example 2 In heart failure cells and animal models, T1AM treatment upregulated SLCO5A1 expression in cardiomyocytes or myocardial tissue.

[0044] The chemical structural formula of T1AM is as follows: Figure 2As shown in A. In the ISO+PE-induced cardiomyocyte hypertrophy model, T1AM treatment upregulated SLCO5A1 protein expression in a concentration-dependent manner ( Figure 2 BC). In mice with heart failure after myocardial infarction, T1AM intervention was found to restore SLCO5A1 protein expression in myocardial tissue, and this effect was dose-dependent. Figure 2 (DE). These results indicate that T1AM, as a pharmacological enhancer of SLCO5A1, can enhance the protein expression of SLCO5A1.

[0045] Example 3 In a mouse model of heart failure, T1AM was found to inhibit myocardial remodeling and improve survival.

[0046] In a mouse model of heart failure following myocardial infarction, T1AM dose-dependently improved mouse survival. Figure 3 A), increasing left ventricular ejection fraction (EF) and shortening left ventricular ejection fraction (FS) ( Figure 3 BC), reduce the expression of myocardial fibrosis marker proteins and myocardial remodeling marker proteins ( Figure 3 DG), reduces cardiomyocyte hypertrophy and decreases myocardial fiber area ( Figure 3 (HK).

[0047] Example 4 In cell models, overexpression of T1AM and SLCO5A1 significantly inhibited the expression of heart failure-related biomarkers in cardiomyocytes, while knockdown of SLCO5A1 significantly enhanced the expression of these related biomarker proteins. Furthermore, T1AM significantly reduced cardiomyocyte hypertrophy, an effect that was reversed by knockdown of SLCO5A1.

[0048] In an ISO+PE-stimulated cardiomyocyte hypertrophy model, T1AM was found to promote SLCO5A1 in a concentration-dependent manner, while reducing the expression of ANP and BNP proteins. Figure 4 Consistent with this, SLCO5A1 overexpression (OE) significantly inhibited ANP and BNP protein expression (AB). Figure 4 Conversely, SCLO5A1 knockdown (Sh) significantly inhibited the regulatory effect of T1AM on these two biomarker proteins (CD). Figure 4 EF). Immunofluorescence assays revealed that T1AM treatment significantly reduced cell area, and this inhibitory effect was reversed by SLCO5A1 knockdown (EF). Figure 4 (GH).

[0049] Example 5 Overexpression of SLCO5A1 in animal models significantly improves myocardial remodeling.

[0050] Using the AAV9-Ctnt-SLCO5A1 vector to mediate cardiomyocyte-specific overexpression of SLCO5A1, tail vein injection of overexpressed (OE) virus or empty viral vector control (Vector) in a mouse model of heart failure revealed that SLCO5A1 overexpression significantly improved left ventricular systolic function. Figure 5 AB), inhibiting the expression of myocardial remodeling marker proteins and fibrosis marker proteins ( Figure 5 The CF, morphologically, also shows that it can alleviate myocardial fibrosis and collagen deposition. Figure 5 GJ).

[0051] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0052] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. The application of an SLCO5A1 overexpression vector in the preparation of products for improving myocardial remodeling, wherein the SLCO5A1 overexpression vector is constructed using adeno-associated virus vector AAV9 and cardiac troponin T as promoters, and the SLCO5A1 overexpression vector is AAV9-Ctnt-SLCO5A1; The SLCO5A1 overexpression vector mediates cardiomyocyte-specific overexpression of SLCO5A1 protein, and the overexpression of SLCO5A1 protein inhibits the expression of myocardial remodeling marker proteins and fibrosis marker proteins.

2. The application according to claim 1, characterized in that, The SLCO5A1 overexpression vector improves left ventricular systolic function, inhibits the expression of myocardial remodeling marker proteins and fibrosis marker proteins, and reduces myocardial fibrosis and collagen deposition by mediating the specific overexpression of SLCO5A1 protein in cardiomyocytes, thereby improving myocardial remodeling.

3. The use of the SLCO5A1 overexpression vector in any one of claims 1-2 in the preparation of a medicament for the treatment and / or prevention of heart failure.

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