Application of substance for increasing content or activity of Nynrin in treatment and / or prevention of cardiovascular diseases

Abstract

The invention discloses application of a substance for increasing the content or activity of Nynrin in treatment and / or prevention of cardiovascular diseases. The research finds that the overexpressed Nynrin can be used for treating and / or preventing cardiovascular diseases. Experiments show that the myocardial cell overexpression Nynrin can reduce the heart injury of mice after myocardial ischemia reperfusion injury, improve the heart function, improve the cardiac fibrosis, and reduce the myocardial infarction area and the myocardial cell death rate. The invention provides theoretical data and experimental support for preparing products for treating and / or preventing cardiovascular diseases.

Description

Technical Field

[0001] This invention is designed for the biomedical field, specifically, to provide the application of a substance that increases the content or activity of Nynrin in the treatment and / or prevention of cardiovascular diseases. Background Technology

[0002] Cardiovascular disease (CVD) is a leading cause of morbidity and mortality worldwide, with ischemic heart disease (IHD) and stroke being the most common types, and myocardial infarction (MI) being its most severe and life-threatening manifestation. Currently, the main clinical treatments for myocardial infarction include percutaneous coronary intervention (PCI) and thrombolytic therapy. However, while these treatments can restore coronary blood flow, the reperfusion process often triggers myocardial ischemia / reperfusion (I / R) injury, further exacerbating initial myocardial damage. The mechanisms of I / R injury are complex, the consequences are severe, and currently, there is a lack of comprehensive and effective treatments and specific drugs for this injury in clinical practice.

[0003] The NYNRIN gene (NYN domain and retroviral integrase containing) is a protein-coding gene located in the q12 region of human chromosome 14, also known as FLJ11811 and CGIN1. The protein encoded by this gene contains the NYN domain and a retroviral integrase-like functional segment, suggesting its potential involvement in key biological processes such as RNA processing, cell cycle regulation, and maintenance of genome stability. NYNRIN is expressed in various tissues, including the heart, brain, liver, kidneys, bone marrow, and testes, with particularly high expression in the nervous and immune systems, suggesting potential tissue-specific functions. However, no studies have yet reported whether NYNRIN can improve cardiovascular disease. Summary of the Invention

[0004] The purpose of this invention is to provide a substance that increases the content or activity of Nynrin for the treatment and / or prevention of cardiovascular diseases.

[0005] The use of Nynrin protein or its active fragments and / or substances that increase the content or activity of Nynrin protein or its active fragments in products for the treatment and / or prevention of cardiovascular diseases.

[0006] The amino acid sequence of the Nynrin protein is SEQ ID NO:1 or its endogenous homologous protein, such as a protein that has 99% identity with the sequence shown in SEQ ID NO:1 and has the same function.

[0007] The substance that increases the content or activity of Nynrin protein or its active fragment is a plasmid containing Nynrin, a recombinant bacterium, a recombinant virus, mRNA, a small molecule agonist, or DNA that can be translated into the Nynrin protein, or an expression cassette, recombinant vector, or recombinant cell containing the DNA.

[0008] Preferably, the nucleotide sequence of the gene encoding the Nynrin protein is SEQ ID NO:2.

[0009] Furthermore, the product for treating and / or preventing cardiovascular diseases is any of the following:

[0010] 1) Products that reduce heart damage;

[0011] 2) Products that improve heart function;

[0012] 3) Products that improve cardiac fibrosis;

[0013] 4) Products that reduce the area of ​​myocardial infarction.

[0014] 5) Products that reduce myocardial cell mortality.

[0015] Furthermore, the cardiovascular diseases include coronary heart disease, hypertension, cardiomyopathy, myocarditis, angina pectoris, coronary atherosclerosis, myocardial infarction, myocardial ischemia-reperfusion injury, valvular heart disease, ischemic heart disease, congenital heart disease, rheumatic heart disease, heart failure, myocardial hypertrophy, myocardial fibrosis, heart failure, arrhythmia, endocarditis, pericarditis, deep vein thrombosis, pulmonary embolism, and pulmonary hypertension.

[0016] Furthermore, the dosage form of the product includes one or more of the following dosage forms: injection, pharmaceutical drops, gel, suspension, emulsion, tablet, capsule, granule, spray, solution, powder, microcapsule, nanoparticle and / or transdermal controlled-release patch.

[0017] Furthermore, the product includes pharmaceuticals.

[0018] A drug for the treatment and / or prevention of cardiovascular disease.

[0019] Furthermore, the drug product for treating and / or preventing cardiovascular diseases has any of the following effects:

[0020] 1) Reduces heart damage;

[0021] 2) Improves heart function;

[0022] 3) Improves cardiac fibrosis;

[0023] 4) Reduce the area of ​​myocardial infarction.

[0024] 5) Reduce myocardial cell mortality.

[0025] Furthermore, the cardiovascular diseases include coronary heart disease, hypertension, cardiomyopathy, myocarditis, angina pectoris, coronary atherosclerosis, myocardial infarction, myocardial ischemia-reperfusion injury, valvular heart disease, ischemic heart disease, congenital heart disease, rheumatic heart disease, heart failure, myocardial hypertrophy, myocardial fibrosis, heart failure, arrhythmia, endocarditis, pericarditis, deep vein thrombosis, pulmonary embolism, and pulmonary hypertension.

[0026] Furthermore, the dosage form of the drug includes one or more of the following dosage forms: injection, pharmaceutical drops, gel, suspension, emulsion, tablet, capsule, granule, spray, solution, powder, microcapsule, nanoparticle and / or transdermal controlled-release patch.

[0027] This invention has discovered that overexpression of Nynrin can treat and / or prevent cardiovascular diseases. For example, overexpression of Nynrin can reduce cardiac damage in mice after myocardial ischemia-reperfusion injury, improve cardiac function, and reduce cardiac fibrosis, myocardial infarction area, and cardiomyocyte mortality, providing theoretical data and experimental support for the preparation of products for the treatment and / or prevention of cardiovascular diseases. Attached Figure Description

[0028] Figure 1 Flowchart for constructing a Nynrin knockout mouse model for cardiomyocytes.

[0029] Figure 2 To validate Nynrin knockout in mouse heart tissue using qPCR.

[0030] Figure 3 To assess cardiac function in mice using echocardiography.

[0031] Figure 4 The ratios of heart weight to body weight and heart weight to tibia length in mice are given.

[0032] Figure 5 Sirius red staining was used to detect the area of ​​cardiac fibrosis in mice.

[0033] Figure 6 TUNEL staining was used to assess cardiomyocyte death in mice.

[0034] Figure 7 TTC staining was used to assess cardiomyocyte death in mice.

[0035] Figure 8 Western blot was used to detect Nynrin overexpression.

[0036] Figure 9To detect myocardial cell damage using a lactate dehydrogenase (LDH) release assay.

[0037] Figure 10 The figures show the in vitro measurements of mitochondrial permeability transition pore (mPTP) opening (top), mitochondrial depolarization (middle), and intracellular reactive oxygen species (ROS) levels (bottom). Detailed Implementation

[0038] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0039] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0040] The detection method in the following embodiment includes the following methods:

[0041] Echocardiography: Mice were anesthetized with 1.5% isoflurane and cardiac function was assessed by transthoracic echocardiography. The heart rate was maintained at 450-550 beats / min during the examination. The measured parameters included left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), left ventricular fraction of shortening (LVFS), and ejection fraction (LVEF).

[0042] Preparation of a mouse model of myocardial ischemia-reperfusion injury: After anesthetizing mice (2% isoflurane), an open-chest surgery was performed at the 4th intercostal space. A silk suture was inserted below the course of the coronary artery to ligate the left anterior descending coronary artery (LAD). The suture was loosened after 40 minutes, and blood perfusion was performed.

[0043] Immunofluorescence staining: After dewaxing and rehydration, paraffin sections were washed three times with PBS, 5 min each time. Sodium citrate antigen retrieval solution was then added to the retrieval jar, and the sections were microwaved on high for 5 min, followed by on low for 15 min. After natural cooling to room temperature, the sections were washed three times with PBS, 5 min each time, and incubated with blocking buffer at room temperature for 1 h. The primary antibody was diluted with antibody diluent and dropped onto the tissue sections, which were then incubated overnight at 4°C. The sections were washed three times with PBS the following day. The corresponding secondary antibody was diluted with antibody diluent, incubated at room temperature for 1 h, and washed three times with PBS. Cell nuclei were stained with Hoechst 33342, incubated at room temperature for 5 min, washed once with PBS, mounted with 50% glycerol, and then photographed and analyzed using a fluorescence microscope.

[0044] Sirius Red Staining: Sirius Red staining was performed using a Sirius Red staining kit (Regan, DC0041). Heart samples were collected, and the heart tissue was immersed in 4% paraformaldehyde (PFA) for dehydration. After paraffin embedding and sectioning (section thickness 5 μm), the tissue was dewaxed, rehydrated, and then stained. For staining, the tissue was immersed in Sirius Red staining solution for 20 minutes, the sections were rinsed, stained with hematoxylin solution for 10 minutes, the slides were rinsed, and washed in xylene. The slides were then mounted with neutral resin mounting medium, air-dried, and observed using a ZeissAxio Observer Z1 microscope. The fibrosis area was measured using Image J.

[0045] TUNEL staining: 24 hours after IR surgery, mice were perfused with physiological saline and euthanized. 20 μg / ml DNase-free proteinase K was added, and the mice were treated at 37°C for 15-30 min, followed by washing three times with PBS (5 min each time). An appropriate amount of TUNEL assay solution was prepared according to the TUNEL kit (Novizan, A112) instructions. The samples were incubated with the TUNEL assay solution at 37°C in the dark for 60 min, followed by washing three times with PBS (5 min each time). Staining was then performed with 0.1-1 μg / ml DAPI at room temperature for 1 min, followed by washing once with PBS (5 min each time). The slides were mounted with 50% glycerol, and then photographed and quantitatively analyzed using a fluorescence microscope.

[0046] TTC staining of mouse hearts: 24 hours after IR surgery, mice were perfused with physiological saline and euthanized. Hearts were then collected and frozen overnight at -80°C. Subsequently, uniform sections of the heart from the ligation site to the apex, 1 mm thick, were prepared and stained with 1% 2,3,5-triphenyltetrazolium chloride (TTC) at 37°C for 15 minutes. Sections were fixed in 4% paraformaldehyde for 24 hours. The infarct area was determined by weighing, by dividing the volume of the infarcted region (pale) by the left ventricular volume measured from the ligation site to the apex.

[0047] Isolation of adult mouse cardiomyocytes: After anesthetizing mice (1.5% isoflurane), the thoracic cavity was opened to expose the heart. The descending aorta was severed, and 3 mL of EDTA buffer was immediately injected into the right ventricle to flush the heart. The ascending aorta was clamped with hemostatic forceps, and the heart was transferred to a culture dish containing fresh EDTA buffer. Then, 10 mL of EDTA buffer, 3 mL of perfusion buffer, and 30-50 mL of collagenase buffer were sequentially injected into the left ventricle for digestion. The heart was then gently pulled into 1 mm fragments with forceps, and 5 mL of stop buffer was used to inhibit enzyme activity. The cell suspension was passed through a 100 μm cell filter, followed by four consecutive rounds of gravity sedimentation. The calcium ion concentration was gradually restored to physiological levels using calcium ion reintroduction buffer. Cardiomyocyte yield and the percentage of viable rod cells were quantified using a hemocytometer. Cardiomyocytes were resuspended in preheated cardiomyocyte culture medium, plated into cell culture plates pre-coated with laminin (5 μg / mL), and cultured in a tissue incubator (37°C, 5% CO2). The culture medium was changed after 24 h, and then every 48 h thereafter.

[0048] In vitro induced oxygen-glucose deprivation / reperfusion (OGD / R): Freshly isolated adult mouse ventricular cardiomyocytes were seeded on laminin-coated 96-well plates for 24 hours to allow adhesion. The culture medium was then replaced with serum-free RPMI1640 (Gibco, 11879020), and the cells were transferred to a hypoxic incubator (1% O2, 94% N2, 5% CO2) for 40 minutes. After this hypoxic period, the serum-free RPMI1640 was replaced with cardiomyocytes, and the cells were returned to a standard incubator for reoxygenation for 24 hours.

[0049] In vitro measurements of mitochondrial permeability transition pore (mPTP) opening, mitochondrial depolarization, and intracellular reactive oxygen species (ROS) levels: Cells were incubated at 37°C for 20 min with 1 μM Calcein-AM (Beyotime, C2012) in the presence of 1 mM cobalt chloride (CoCl2), 200 nM tetramethylrhodamine ethyl ester (TMRE), or 5 μM CellROX Green Reagent. Specifically, mPTP opening was assessed by measuring the green fluorescence intensity (488 nm) of Calcein-AM; mitochondrial membrane potential was assessed by measuring the red fluorescence intensity (561 nm) of TMRE; and intracellular ROS levels were determined by measuring the fluorescence intensity (488 nm) of the CellROX Green Reagent. Imaging was performed using an Andor Dragonfly 505 confocal microscope, and quantitative analysis was performed using ImageJ software.

[0050] Lactate dehydrogenase (LDH) release assay: Cell death is assessed by detecting the activity of lactate dehydrogenase (LDH) released from cultured cells. Cells are seeded in 96-well plates, and the supernatant is collected. The LDH activity in the supernatant is determined spectrophotometrically using an LDH cytotoxicity assay kit (Yeasen, 40209ES76) according to the manufacturer's instructions.

[0051] Western blotting: using Quick Start TM Protein concentration was quantified using the Bradford kit (Beyotime, P0006). Each sample was subjected to SDS-PAGE and electrotransfer to a PVDF membrane, then blocked with 5% skim milk, incubated with primary antibody, and then incubated with horseradish peroxidase (HRP)-labeled secondary antibody. Statistical analysis was performed using the Odyssey-Fc system.

[0052] Real-time quantitative PCR (qPCR): Total RNA was extracted from heart tissue using TRIzol reagent (Thermo Fisher, 15596018). RevertAid was used. TM The premixed buffer (Thermo Fisher, M1632) was reverse-engineered into cDNA. qPCR data were normalized using the internal control gene GAPDH.

[0053] Example 1: Construction of Nynrin-specific knockout mice for cardiomyocytes

[0054] 1. Construction of Nynrin-specific knockout mice for cardiomyocytes

[0055] This example demonstrates the construction of cardiomyocyte-specific Nynrin knockout mice. We first introduced Nynrin... fl / fl Mice (gifted by Professor Hou Yu of Chongqing Medical University, see published article https: / / doi.org / 10.1016 / j.stem.2024.06.007) were mated with Myh6-MerCreMer mice (purchased from The Jackson Laboratory, strain number JAX005657) to establish a tatamoxifen-induced cardiomyocyte-specific Nynrin knockout (Nynrin-cKO) mouse model. Figure 1 Mice were injected with tamoxifen (40 mg / kg) for 5 consecutive days at 5 weeks of age.

[0056] 2. Verification Results

[0057] Mice from step 1 were collected at 8 weeks of age for qPCR analysis. The results showed that Nynrin was specifically knocked out in the heart tissue of mice at 8 weeks of age. Figure 2 ).

[0058] Example 2: Cardiac cell-specific knockout of Nynrin exacerbates cardiac dysfunction and cardiac injury in mice with myocardial ischemia-reperfusion injury.

[0059] This embodiment uses a myocardial ischemia-reperfusion injury model as an example to explore how cardiomyocyte-specific knockout of Nynrin exacerbates cardiac dysfunction and cardiac injury after cardiovascular disease. A mouse model of Nynrin-specific knockout in cardiac tissue was constructed according to the method in Example 1. Mice in the Nynrin (Ctrl group) and Nynrin-cKO (KO group) groups underwent myocardial ischemia-reperfusion (I / R) surgery, and echocardiography was performed one week after surgery to assess cardiac function. Echocardiographic assessment one week after I / R showed that cardiac function in Nynrin-cKO mice was significantly worse, with increased LVEDD and LVESD, and decreased LVEF and LVFS compared to the Ctrl group mice. Figure 3 ).

[0060] Subsequently, samples were collected and weighed. The ratio of heart weight to body weight (HW / BW) and the ratio of heart weight to tibia length (HW / TL) in Nynrin-cKO mice were also significantly increased. Figure 4 ).

[0061] In addition, Sirius red staining was performed to assess cardiac fibrosis in mice. The Sirius red staining results further showed that, compared with the Ctrl group mice, the Nynrin cKO mice had an increased area of ​​cardiac fibrosis scarring. Figure 5 The results indicate that the KO group mice suffered more severe cardiac damage. These data suggest that cardiomyocyte-specific knockout of Nynrin leads to myocardial fibrosis and cardiac dysfunction.

[0062] Twenty-four hours after I / R, samples were collected for TUNEL and TTC staining to assess cardiomyocyte death in mice. TUNEL staining results showed that 24 hours after I / R, the cardiomyocyte mortality rate in the Nynrin-cKO cardiac peripheral zone was significantly higher than that in the Ctrl group. Figure 6 Furthermore, triphenyltetrazolium chloride (TTC) staining showed that Nynrin-cKO mice had a larger area of ​​myocardial infarction tissue compared to Ctrl mice. Figure 7 In summary, these data indicate that Nynrin-cKO increases cardiomyocyte death in mice after I / R.

[0063] Example 3: Nynrin overexpression alleviates cardiomyocyte injury under oxygen-glucose deprivation / reperfusion (OGD / R) conditions.

[0064] Lentiviral vector construction: using pCDH Nynrin plasmid (gifted by Professor Hou Yu of Chongqing Medical University, see the published article https: / / doi.org / 10.1016 / j.stem.2024.06.007) and lentivirus packaging in 293T cells, Nynrin-overexpressing lentiviruses were obtained.

[0065] We isolated primary adult mouse cardiomyocytes and overexpressed Nynrin in them using a lentiviral vector to evaluate its potential beneficial effects on cardiomyocytes under OGD / R conditions. Western blot analysis confirmed successful overexpression of Nynrin (Thermo Fisher, PA553881) in cardiomyocytes. Figure 8 LDH release assays showed that, compared with the Ctr1 group (luciferase overexpression), Nynrin-overexpressing cardiomyocytes exhibited significantly reduced damage. Figure 9 ).

[0066] Furthermore, our analysis showed that Nynrin overexpression significantly increased Calcein-AM fluorescence in the presence of CoC12. Figure 10 The image above shows that OGD / R conditions prevent mPTP opening. Furthermore, Nynrin overexpression also increases TMRE fluorescence (…). Figure 10 (middle image) and reduced Cel1ROX fluorescence ( Figure 10 The figure below shows that Nynrin overexpression alleviates mitochondrial depolarization and reduces oxidative stress levels in adult mouse cardiomyocytes. In summary, these data indicate that Nynrin overexpression mitigates cardiomyocyte injury under oxygen-glucose deprivation / reperfusion (OGD / R) conditions.

[0067] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some basic features can be applied within the scope of the following appended claims. SEQ ID NO:1: Nynrin amino acid sequence (1840aa):

[0068] MLLSGGKPPAQEWFMVQTKSKPRVHRQRLQVQRIFRVKVTAFQSRPDTPYFW

[0069] LQLEGPRENTGKAKEYLKGLCNPELWKEVRYPPLHLCAFLGAQGLFLDCLCW

[0070] STLAYLVPGPPGSLMVGGLTESFTMTQNWLEELVARLRWGPAPMITPRGVWET

[0071] EVTRAFGALVWIRCDKYAGDLLQLPPAVQELLLSLVRDAAGKEDIIEWLGHFGI

[0072] SGTCPNPEILICLARQQKESASLVTIKESPGTLQEIGALNRASENSKKTTSSGAA

[0073] GSLTQAQSPPAQETADQLARDQSNKQGDETNSVGEEGTATQDTSSQDSENPTQ

[0074] ALLQQKQVPKNEERISLLLPVSALSAYTSWKVWAPGTAFGPSWPGTIAATFWK

[0075] INELQSLHLAWLLSQACNLNFFWQRPTGPIQLKLPGRNPLPLKLEWKQKELVPL

[0076] SSAGSPACRPGGDLGRETALKHSPRPEIPSKIISLSVVPGGCGIKEKVSPGLLQV

[0077] GQSSTSVGDKGISLSDCKGLEKPFSLALSTEQGGSTAQERPLAQVPEAPTVSET

[0078] LQVATAAEVSNVEHPPTGEGLPATPKVPTALKKPAVYTEPTAPKVPSAPTEPAAP

[0079] ATPTAPQTPTAQKTPSVKTLAGLQTPKVQSETIATAGSEVPKAPAASAVAGPTV

[0080] DVAQLLSEVQASKNRAIMLKVQGKPGRQGFQPSTVPSRSKHQFLKEGLLGA

[0081] WEGSQRLSPHSQGTNIVTSFQRYNEALNTPFEMNLSEEPGNPGLRRVVIDGSSV

[0082] AMVHGLQHFFSCRGIAMAVQYFWNRGHREITVFVPTWQLKKNRRVRESHFLT

[0083] KLHRLKMLSITPSQLENGKKITTYDYRFMVKLAEETDGVIVTNEQIHILMNNS

[0084] KKLMVKDRLLPFTFAGSLFMVPDDPLGRDGPTLEEFLKKPNRLDMDIGNFLKV

[0085] WKTLPPSSASISELSDDADAEPLEDPQDVEEAGKEEGSLEEEPGIPKPDEEDEQ

[0086] DTNPVSVFGVECPSFSEEILQCLSLHDPSEGTLDIDLLPVVSSPYLDVPWDGKA

[0087] PCQQVLAQLAQLNIPSNFTALSFFMGFMDSHRDVISDYEDLVGPLHGLLKQKP

[0088] DWQWNQEHEKSFLALKRALVCALCLSTPNPNLPFYLEVTVSQVSLTASLHQEH

[0089] SGRKHPIAYTSKPLLPDEDSEGPQSGGDSPYAVAWALKHFARCVGDNPVVLRLS

[0090] YASRTTVDNEAWDSRRASKAWLIRWSLLLQDKGKRELELSLLQGLLGENQLLT

[0091] PPSSMPRVFQPLPPSSDLSTFICVHVSGYCFYRDDELCAGFGLYILSPTSPPVSLA

[0092] FSCSPYTTTYAHLAAVACGLERFGQSQHPVVFLTHCNWIFSVLWELLPLWRVR

[0093] GFLSSDGASLPHPSLLSYIISLTSGFSPLPFIYRTSYRGSLFAVTVDTLAKQGAQG

[0094] GGQWWDLPKDVPVPMVTPHPKGRKPNLLALQLSDTTLADIIAKLQAGQKLSG

[0095] PSPFSSAFNSLSLDQDSGLLMFKGERHPRVWVVPRQLRRDLIFSVHDSPIGEHQ

[0096] GLEDTYKTVRLLGWWPGMQDHVRDYCRSCLFCIPRNLIGGELKVIESPWPLRS

[0097] TAPWSSLQIEVVGPVTVSEEGHKHVLIVADANTRWVEAFPLKPYTHVAVAQVL

[0098] LQHVFARWGVPIRLEAAQGPQFARHVLVSCGLALGAQVTTLSRALQFPCLMSS

[0099] EAYWEFKRALKEFIFLYGKKWAASLPLLHLAFRASTTEATPFQVLTGGEMKLM

[0100] EPVWWEMSRANIEGLKMDAFLLQLMRELLDLHWRVAEKASEKAENRRFKRE

[0101] SQENEWSVGDQVLLLSLPRNGSSAKWMGPFYIGDRLSLSLYRVWGFPVPDKL

[0102] GCVYPSSLMKAFPKHDTPLSLDVEQ

[0103] SEQ ID NO:2: Nynrin CDS fragment (5523bp):

[0104] ATGCTTTTGTCGGGGGGTAAACCCCCAGCGCAGGAATGGTTCATGGTGCAG

[0105] ACAAAATCGAAGCCCCGGGTACACCGGCAGCGGCTGCAAGTCCAGCGCAT

[0106] CTTCAGGGTCAAGGTGACCGCTTTCCAGAGCCGTCCTGACACCCCTTACTT

[0107] TTGGCTGCAGCTGGAGGGGCCCCGGGAAAATACGGGCAAAGCCAAGGAAT

[0108] ATCTGAAAGGTCTGTGCAACCCAGAGCTATGGAAGGAGGTTCGCTACCCAC

[0109] CAGTCCTGCACTGCGCCTTCCTTGGGGCACAAGGCTTGTTCCTGGACTGCC

[0110] TCTGCTGGAGTACCCTGGCCTACCTGGTGCCTGGTCCCCCTGGCTCCTTAAT

[0111] GGTGGGTGGGCTGACCGAGTCTTTTACCATGACACAGAACTGGCTGGAGGA

[0112] GCTGGTGGCACGGTTGCGCTGGGGCCCTGCCCCAATGATCACCCCTCGAGG

[0113] GGTTTGGGAGACAGAGGTGACCCGGGCTTTTGGGGCTCTGGTGTGGATCCG

[0114] TTGTGACAAGTATGCAGGAGACTTGCTGCAGCTTCCTCCAGCAGTCCAGGA

[0115] GCTGCTTCTCAGTTTGGTCCGAGATGCTGCCGGCAAGGAAGACATCATTGA

[0116] GTGGCTCGGCCATTTTGGCATCTCTGGTACTTGCCCCAACCCAGAGATCCTG

[0117] ATCTGCCTTGCCCGGCAGCAGAAGGAAAGCGCATCCCTGGTGACCATAAAA

[0118] GAGAGCCCCGGGACGCTCCAAGAGATAGGAGCCTTGAATCGGGCTTCAGA

[0119] AAATTCAAAGAAAACAACAAGTTCGGGAGCCGCTGGGTCCCTGACCCAAG

[0120] CTCAGAGCCCTCCGGCGCAGGAGACAGCAGACCAGCTAGCACGGGACCAG

[0121] TCTAACAAGCAAGGTGACGAAACGAACAGTGTTGGAGAGGAAGGGACCGC

[0122] TACACAAGACACCAGCAGCCAGGACTCTGAGAATCCCACACAAGCCCTCTT

[0123] GCAGCAAAAGCAGGTACCGAAGAATGAAGAGAGAATCTCATTGCTGTTGCC

[0124] AGTGTCAGCTCTGAGCGCGTACACATCCTGGAAGGTCTGGGCCCCTGGGAC

[0125] AGCCTTTGGGCCCTCGTGGCCAGGGACCATTGCTGCCACCTTCTGGAAGAT

[0126] CAATGAACTGCAGTCTCTTCACCTAGCCTGGCTCCTGTCCCAGGCTTGCTTA

[0127] AATTTCCCCTTCTGGCAGAGGCCCACGGGCCCCATTCAGCTGAAGCTGCCA

[0128] GGACGGAATCCTTTGCCCTTAAAGCTGGAATGGAAGCAGAAAGAGCTGGTT

[0129] CCTCTGTCCAGTGCAGGAAGCCCAGCTTGTAGACCAGGTGGGGACCTAGG

[0130] GAGGGAGACAGCCCTAAAGCATAGCCCGAGGCCGGAGATTCCCTCCAAAAT

[0131] CATCAGTTTATCGGTGGTGCCAGGGGGTTGTGGTATCAAAGAGAAGGTTAG

[0132] CCCGGGACTTCTGCAAGTAGGGCAGTCTTCAACATCTGTGGGAGATAAAGG

[0133] AATTTCACTGTCAGATTGTAAAGGCCTGGAAAAGCCGTTCTCTCTAGCACTG

[0134] TCCACAGAGCAAGGGGGATCCACAGCTCAAGAGAGGCCACTGGCTCAAGT

[0135] ACCCGAAGCTCCAACGGTGTCTGAAACTCTTCAAGTGGCCACGGCTGCAG

[0136] AGGTGTCCAACGTTGAACACCCACCCACCGGGGAGGGGCTGCCTGCAACC

[0137] CCCAAAGTGCCTACAGCCCTAAAGAAGCCAGCAGTGTACACAGAGCCCAC

[0138] AGCTCCCAAAGTGCCTTCGGCTCCAACAGAACCAGCAGCTCCTGCCACGCC

[0139] CACAGCCCCACAAACTCCTACAGCTCAGAAAACACCTTCAGTGAAAACAC

[0140] TCGCAGGCCTCCAGACCCCCAAAGTTCAAAGTGAGACCATAGCTACAGCAG

[0141] GGTCTGAAGTTCCCAAAGCGCCTGCAGCTTCAGCAGTTGCGGGGCCGACC

[0142] GTGGATGTAGCCCAACTCCTGAGTGAGGTTCAGGCTTCAAAGAATAGAGCC

[0143] ATCATGTTGAAGGTCCAGGGAAAGCCCGGAAGGCAGGGTTTCCAGCCCAGT

[0144] AGCACCGTGCCCTCCAGAAGTAAGCATCAGTTCCTAAAGGAAGGACTTCTT

[0145] GGGGCTTGGGAGGGGTCCCAGAGGCTGTCACCTCACTCCCAGGGCACCAA

[0146] CATAGTGACCAGCTTCCAGAGGTACAACGAGGCCTTGAACACACCGTTCGA

[0147] GATGAACTTGTCTGAGGAGCCTGGAAATCCAGGGCTGAGGAGAGTGGTCAT

[0148] TGACGGCAGCAGTGTGGCCATGGTACATGGCCTGCAGCACTTCTTCTCGTG

[0149] CCGAGGCATCGCCATGGCCGTGCAGTACTTCTGGAATCGCGGACACCGAGA

[0150] GATCACCGTGTTCGTACCCACCTGGCAGCTGAAGAAAAACCGGAGGGTGA

[0151] GAGAGAGCCACTTTCTGACGAAGCTTCACAGACTGAAGATGCTGTCTATCA

[0152] CTCCCTCGCAGCTGGAGAATGGCAAAAAGATCACCACTTACGATTACAGGT

[0153] TCATGGTAAAGCTGGCAGAGGAGACAGATGGAGTCATTGTCACCAATGAGC

[0154] AGATCCACATCCTCATGAATAATTCCAAGAAACTGATGGTCAAAGATCGCCT

[0155] GCTGCCCTTCACCTTTGCCGGGAGTCTCTTTATGGTACCAGATGACCCCCTG

[0156] GGCCGTGACGGCCCCACCTTGGAGGAATTTCTGAAGAAGCCAAACAGGTT

[0157] GGACATGGACATTGGCAATTTCTTGAAGGTGTGGAAGACCCTGCCTCCCAG

[0158] CTCAGCCAGCATCTCTGAACTGAGTGATGACGCTGACGCCGAACCTTTGGA

[0159] GGATCCACAGGACGTAGAAGAAGCAGGGAAGGAGGAGGGAAGCCTGGAA

[0160] GAGGAGCCAGGAATCCCAAAGCCCGATGAAGAAGATGAGCAGGACACTAA

[0161] CCCTGTGTCGGTGTTTGGGGTCGAGTGCCCTTCATTTTCCGAGGAAATCCTC

[0162] CAGTGTCTCAGCCTGCATGACCCCTCTGAGGGGACTTTAGACATCGACCTTC

[0163] TGCCTGTGGTGTCCTCTCCGTACCTGGATGTCCCCTGGGATGGGAAGGCTCC

[0164] CTGTCAGCAGGTTCTGGCCCAGCTGGCGCAGCTTAACATCCCGAGCAACTT

[0165] CACCGCGCTATCCTTCTTCATGGGCTTCATGGATTCCCACCGGGATGTTATCT

[0166] CTGACTATGAAGATCTCGTGGGCCCCCTGCACGGCCTCCTCAAGCAGAAGC

[0167] CGGACTGGCAGTGGAACCAGGAGCACGAGAAGTCCTTCCTGGCCCTGAAA

[0168] CGAGCCCTAGTGTGCGCCCTCTGCCTGTCAACCCCCAACCCCAACCTGCCC

[0169] TTCTACCTGGAAGTGACTGTCAGCCAAGTGTCACTGACAGCTAGCTTGCAC

[0170] CAGGAGCACTCAGGAAGGAAGCACCCCATCGCTTATACCTCGAAACCTCTC

[0171] CTCCCCGATGAAGACAGCGAGGGTCCTCAGTCAGGGGGGGACAGCCCCTAT

[0172] GCTGTGGCTTGGGCCCTCAAGCATTTTGCCCGATGCGTTGGAGACAATCCA

[0173] GTGGTTCTACGTCTTTCCTATGCCTCCCGGACCACGGTGGATAATGAGGCAT

[0174] GGGATAGCCGTAGGGCTTCGAAAGCGTGGTTGATTCGGTGGTCTCTCTTGCT

[0175] GCAGGACAAAGGCAAGAGGGAATTGGAATTGTCCCTTCTCCAGGGCCTGCT

[0176] GGGGGAGAACCAGCTGCTGACACCCCCTTCCTCCATGCCCCGAGTTTTCCA

[0177] GCCTCTGCCTCCTTCTTCTGACCTGTCTACTTTTATCTGTGTCCATGTGTCTG

[0178] GCTATTGCTTCTACCGTGACGATGAGTTGTGTGCTGGCTTTGGTCTCTACATC

[0179] TTGTCGCCCACCAGCCCTCCAGTCTCCCTTGCCTTTTCCTGTTCTCCTTACAC

[0180] AACCACATACGCCCACCTGGCAGCCGTGGCCTGCGGCTTAGAGCGCTTCGG

[0181] CCAGTCCCAGCACCCTGTGGTTTTCCTCACCCACTGCAACTGGATCTTCAGC

[0182] GTTCTCTGGGAGCTCCTGCCCCTCTGGAGAGTCCGGGGCTTCCTGTCATCTG

[0183] ACGGGGCTTCACTACCTCATCCAAGCCTGCTGTCCTACATCATCTCTCTCAC

[0184] GTCTGGCTTTTCACCCCTTCCCTTTATCTACCGAACCTCTTATCGGGGCTCTC

[0185] TGTTTGCTGTGACAGTGGACACTCTGGCCAAGCAGGGTGCCCAAGGGGGC

[0186] GGGCAGTGGTGGGATTTGCCAAAGGATGTGCCAGTGCCAATGGTGACTCCC

[0187] CATCCTAAAGGCAGGAAGCCCAACTTGCTGGCCTTACAGCTGAGTGATACC

[0188] ACCCTGGCCGATATCATTGCCAAGTTGCAGGCAGGACAGAAATTGTCCGGG

[0189] CCCTCCCCTTTCAGTTCTGCCTTTAACTCACTCAGCCTGGACCAAGACAGTG

[0190] GCCTGCTCATGTTCAAGGGGGAAAGGCATCCCAGGGTTTGGGTAGTCCCAA

[0191] GGCAACTTCGGAGGGATCTGATTTTCTCTGTGCATGACAGCCCCATTGGGG

[0192] AACACCAGGGGCTAGAGGACACCTATAAGACGGTGAGGCTGCTGGGATGG

[0193] TGGCCTGGGATGCAGGACCACGTGAGAGATTACTGCAGGAGCTGTTTGTTC

[0194] TGCATCCCCCGGAATCTCATAGGCGGGGAGCTGAAAGTTATCGAGTCCCCAT

[0195] GGCCTCTCAGGTCGACAGCCCCTTGGTCCAGTCTGCAGATTGAGGTGGTGG

[0196] GTCCAGTCACTGTAAGTGAGGAGGGACATAAGCACGTGCTCATTGTGGCCG

[0197] ATGCCAACACCCGGTGGGTGGAGGCATTCCCTCTGAAGCCCTATACGCACG

[0198] TGGCTGTGGCCCAGGTGCTCCTCCAGCATGTGTTTGCAAGGTGGGGCGTTC

[0199] CCATCAGACTGGAGGCCGCCCAAGGCCCCCAGTTTGCCCGCCATGTTCTGG

[0200] TGAGCTGTGGGCTGGCCCTGGGAGCCCAAGTGACTACACTGAGTAGGGCC

[0201] CTCCAGTTCCCCTGCCTGATGAGTTCAGAGGCCTACTGGGAATTCAAGAGG

[0202] GCCCTGAAGGAGTTCATCTTTCTGTATGGCAAGAAGTGGGCGGCCTCTCTG

[0203] CCCTTGCTGCACCTGGCTTTTAGGGCCTCCACCACAGAGGCCACACCGTTC

[0204] CAGGTGCTGACCGGGGGCGAGATGAAGCTGATGGAGCCCGTGTGGTGGGA

[0205] GATGAGCCGGGCGAACATTGAAGGGCTCAAGATGGACGCTTTCTTGCTGCA

[0206] GCTGATGCGGGAGCTGTTGGACCTCCACTGGAGAGTGGCCGAGAAGGCCA

[0207] GCGAGAAGGCCGAGAACAGGCGTTTCAAGAGGGAGAGCCAGGAGAACGA

[0208] GTGGAGTGTGGGCGACCAGGTCCTCTTGTTGTCTCTCCCCAGGAATGGCAG

[0209] CAGTGCCAAATGGATGGGTCCTTTCTATATTGGGGACCGGTTGAGCCTATCA

[0210] CTCTATAGGGTGTGGGGCTTCCCAGTCCCAGACAAGCTGGGCTGTGTCTATC

[0211] CTAGCAGTCTGATGAAGGCCTTTCCCAAGCATGACACGCCCCTGTCCCTCG

[0212] ACGTGGAGCAGTGA。

Claims

1. The use of Nynrin protein or its active fragments and / or substances that increase the content or activity of Nynrin protein or its active fragments in products for the treatment and / or prevention of cardiovascular diseases.

2. The application according to claim 1, characterized in that, The substance that increases the content or activity of Nynrin protein or its active fragment is a plasmid containing Nynrin, a recombinant bacterium, a recombinant virus, mRNA, a small molecule agonist, or DNA that can be translated into the Nynrin protein, or an expression cassette, recombinant vector, or recombinant cell containing the DNA.

3. The application according to any one of claims 1-2, characterized in that, The product for treating and / or preventing cardiovascular disease is any of the following: 1) Products that reduce heart damage; 2) Products that improve heart function; 3) Products that improve cardiac fibrosis; 4) Products that reduce the area of ​​myocardial infarction; 5) Products that reduce myocardial cell mortality.

4. The application according to any one of claims 1-3, characterized in that, The cardiovascular diseases mentioned include coronary heart disease, hypertension, cardiomyopathy, myocarditis, angina pectoris, coronary atherosclerosis, myocardial infarction, myocardial ischemia-reperfusion injury, valvular heart disease, ischemic heart disease, congenital heart disease, rheumatic heart disease, heart failure, myocardial hypertrophy, myocardial fibrosis, heart failure, arrhythmia, endocarditis, pericarditis, deep vein thrombosis, pulmonary embolism, and pulmonary hypertension.

5. The application according to any one of claims 1-4, characterized in that, The dosage forms of the product include one or more of the following: injection, pharmaceutical drops, gel, suspension, emulsion, tablet, capsule, granule, spray, solution, powder, microcapsule, nanoparticle and / or transdermal controlled-release patch.

6. The application according to any one of claims 1-5, characterized in that, The products include pharmaceuticals.

7. A drug for the treatment and / or prevention of cardiovascular disease.

8. The medicament according to claim 7, characterized in that, The medication for treating and / or preventing cardiovascular disease has any of the following effects: 1) Reduces heart damage; 2) Improves heart function; 3) Improves cardiac fibrosis; 4) Reduce the area of ​​myocardial infarction; 5) Reduce myocardial cell mortality.

9. The medicament according to any one of claims 7-8, characterized in that, The cardiovascular diseases mentioned include coronary heart disease, hypertension, cardiomyopathy, myocarditis, angina pectoris, coronary atherosclerosis, myocardial infarction, myocardial ischemia-reperfusion injury, valvular heart disease, ischemic heart disease, congenital heart disease, rheumatic heart disease, heart failure, myocardial hypertrophy, myocardial fibrosis, heart failure, arrhythmia, endocarditis, pericarditis, deep vein thrombosis, pulmonary embolism, and pulmonary hypertension.

10. The medicament according to any one of claims 7-9, characterized in that, The dosage form of the drug includes one or more of the following: injection, pharmaceutical drops, gel, suspension, emulsion, tablet, capsule, granule, spray, solution, powder, microcapsule, nanoparticle and / or transdermal controlled-release patch.

Images