Application of mitochondrial pyruvic acid transporter 1 and / or detection reagent thereof in preparation of coronary heart disease detection product

Abstract

The invention relates to the field of biomedicine and molecular biology, in particular to application of a mitochondrial pyruvic acid transporter 1 and / or a detection reagent thereof in preparation of coronary heart disease detection products. Compared with the traditional technology, the application discovers and proposes for the first time that the MPC1 specifically expressed in the circulating exosome is used as the diagnosis biomarker of the coronary heart disease, and a new scheme is provided for clinical detection of the coronary heart disease.

Description

Technical Field

[0001] The present application relates to the fields of biomedicine and molecular biology, and in particular to the use of a mitochondrial pyruvate transporter 1 and / or a detection reagent thereof in the preparation of a coronary heart disease detection product. Background Art

[0002] With the increasing incidence of chronic diseases such as hypertension, hyperlipidemia, and hyperglycemia, as well as the increasing aging of the population, coronary heart disease (CAD) has become one of the leading causes of death and disability worldwide. Exploring the causes and target molecules of CAD is crucial to improving diagnostic accuracy and enhancing treatment efficacy.

[0003] Coronary heart disease is a complex multifactorial disease. Currently, the common biomarkers in clinical practice mainly include:

[0004] (1) Troponin (Troponin T and Troponin I): Troponin is a highly specific marker of myocardial cell damage. In recent years, high-sensitivity troponin (hs-TNT) has been considered the "gold standard" for diagnosing acute coronary heart disease (especially acute myocardial infarction, AMI). The level of troponin in the blood increases significantly after myocardial ischemia and necrosis. Troponin is almost only present in myocardial cells and is highly sensitive to myocardial damage. Continuous monitoring of the rising and falling trends of troponin levels can help determine the time window of acute myocardial infarction and the risk of further myocardial damage.

[0005] (2) Creatine kinase isoenzyme (CK-MB): CK-MB is traditionally used to detect acute myocardial infarction. It mainly reflects the release of enzymes after myocardial cell damage. It has a strong correlation with the myocardium and can quickly reflect myocardial damage.

[0006] (3) C-Reactive Protein (CRP): CRP is a sensitive marker of inflammatory response and is used to assess the inflammatory status of the coronary arteries and the overall risk of cardiovascular disease. Elevated CRP is closely related to the progression of atherosclerosis in coronary heart disease and can be used to predict the risk of cardiovascular events (such as myocardial infarction, ischemic stroke, etc.).

[0007] (4) Brain natriuretic peptide (BNP) and amino-terminal pro-brain natriuretic peptide (NT-proBNP): These two markers are mainly used to assess heart failure, but they also show high levels in conditions of myocardial stress and decreased cardiac function caused by coronary heart disease. They can reflect cardiac load and volume overload and are of great significance in assessing the risk of coronary heart disease complicated by heart failure.

[0008] The discovery of new biomarkers is of great significance for the clinical detection of coronary heart disease. Summary of the invention

[0009] Based on this, one or more embodiments of the present application provide the use of a detection reagent for mitochondrial pyruvate carrier 1 (MPC1) protein in the preparation of a coronary heart disease detection product. The following technical solutions are included:

[0010] One or more embodiments of the present application provide a use of a mitochondrial pyruvate transporter 1 and / or a detection reagent thereof in the preparation of a coronary heart disease detection product.

[0011] In some embodiments of the present application, the sample detected by the coronary heart disease detection product includes body fluids of the circulatory system.

[0012] In some embodiments of the present application, the sample detected by the coronary heart disease detection product includes blood.

[0013] In some embodiments of the present application, the coronary heart disease detection product detects exosome mitochondrial pyruvate transporter 1 in a sample.

[0014] In some embodiments of the present application, the coronary heart disease detection product also includes one or more of an exosome extraction reagent and an exosome detection reagent.

[0015] In some embodiments of the present application, the detection reagent in the coronary heart disease detection product is detected by any of the following methods: mass spectrometry, immunoassay, western blot analysis, radioimmunoassay, immunofluorescence, immunoprecipitation, equilibrium dialysis, immunodiffusion, electrochemiluminescence immunoassay, ELISA assay and immunopolymerase chain reaction.

[0016] In some embodiments of the present application, the coronary heart disease detection product is a detection kit or a chip.

[0017] In some embodiments of the present application, the coronary heart disease detected by the coronary heart disease detection product includes acute myocardial infarction.

[0018] In some embodiments of the present application, the coronary heart disease detected by the coronary heart disease detection product includes myocardial necrosis.

[0019] In some embodiments of the present application, the coronary heart disease detected by the coronary heart disease detection product includes acute coronary syndrome.

[0020] One or more embodiments of the present application provide a coronary heart disease detection product as defined above.

[0021] Compared with traditional technologies, this application discovered and proposed for the first time the use of MPC1 specifically expressed in circulating exosomes as a diagnostic biomarker for coronary heart disease, providing a new solution for the early detection of clinical coronary heart disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the technical solutions in the embodiments of the present application and to more completely understand the present application and its beneficial effects, the following is a brief introduction to the drawings required for the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0023] Figure 1 The WB identification of plasma exosomes and the ELISA determination of MPC1 expression results in the established mouse acute myocardial infarction model;

[0024] Figure 2 Western blot was used to detect the expression of TSG101, CD9, and GM130 proteins in plasma exosomes of clinical patient samples;

[0025] Figure 3 Correlation and ROC curve analysis results. DETAILED DESCRIPTION

[0026] The present application will be further described in detail below in conjunction with the accompanying drawings, embodiments and examples. It should be understood that these embodiments and examples are only used to illustrate the present application and are not used to limit the scope of the present application. The purpose of providing these embodiments and examples is to make the understanding of the disclosure of the present application more thorough and comprehensive. It should also be understood that the present application can be implemented in many different forms, is not limited to the embodiments and examples described herein, and those skilled in the art can make various changes or modifications without violating the connotation of the present application, and the equivalent form obtained also falls within the protection scope of the present application. In addition, in the description below, a large number of specific details are given in order to provide a more comprehensive understanding of the present application, and it should be understood that the present application can be implemented without one or more of these details.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing implementation modes and embodiments and are not intended to limit this application.

[0028] the term

[0029] Unless otherwise specified or incompatible herewith, the terms and phrases used herein shall have the following meanings:

[0030] The terms "and / or", "or / and", and "and / or" used in this article include any one of two or more related listed items, and also include any and all combinations of related listed items, and the arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution that is all connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the combination of four of A, B, C, and D (that is, the technical solution that is all connected by "logical AND").

[0031] In the present application, "plurality", "multiple", "multiple times", "multiples", etc., unless otherwise specified, refer to a number greater than 2 or equal to 2. For example, "one or more" means one or greater than or equal to two.

[0032] As used herein, "combination thereof", "any combination thereof", "any combination thereof" etc. include all suitable combinations of any two or more of the listed items.

[0033] Herein, the “suitable” mentioned in “suitable combination”, “suitable method”, “any suitable method”, etc., shall be based on the ability to implement the technical solution of this application, solve the technical problems of this application, and achieve the expected technical effects of this application.

[0034] Herein, “preferred”, “better”, “more preferred” and “suitable” are merely used to describe implementation methods or examples with better effects, and it should be understood that they do not constitute limitations on the scope of protection of this application.

[0035] In the present application, "further", "further", "particularly" and the like are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0036] In this application, "optionally", "optional", and "optional" mean optional or dispensable, that is, any one of the two parallel schemes of "yes" or "no". If multiple "options" appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent.

[0037] In the present application, the terms "first", "second", "third", "fourth", etc. in "the first aspect", "the second aspect", "the third aspect", "the fourth aspect", etc. are used only for descriptive purposes and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity.

[0038] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0039] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as t is an integer selected from 1 to 10, indicating that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be combined. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges included therein.

[0040] The temperature parameters in this application, unless otherwise specified, are allowed to be either constant temperature treatment or to vary within a certain temperature range. It should be understood that the constant temperature treatment allows the temperature to fluctuate within the accuracy range controlled by the instrument. Fluctuations within the range of ±5°C, ±4°C, ±3°C, ±2°C, and ±1°C are allowed.

[0041] In the present application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass volume percentage.

[0042] All documents mentioned in this application are cited as references in this application, just as each document is cited as reference separately. Unless they conflict with the application purpose and / or technical solution of this application, the cited documents involved in this application are cited with all contents and all purposes. When the cited documents are involved in this application, the definitions of relevant technical features, terms, nouns, phrases, etc. in the cited documents are also cited. When the cited documents are involved in this application, the examples and preferred methods of the cited relevant technical features can also be incorporated into this application as references, but are limited to the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, the present application shall prevail or be modified adaptively according to the description of this application.

[0043] At present, the biomarkers of coronary heart disease mainly include:

[0044] (1) Troponin. Elevated troponin is usually associated with acute myocardial injury and is therefore mainly used for the diagnosis of acute coronary syndrome (ACS).

[0045] (2) CK-MB. CK-MB may also increase in skeletal muscle injury, extensive exercise, or other kidney diseases, so its specificity is not as good as troponin.

[0046] (3) CRP: CRP is a nonspecific marker of inflammation and may be elevated due to other inflammatory diseases (such as infection and rheumatic diseases). CRP levels cannot be used alone to diagnose coronary heart disease, but can be used as an auxiliary predictive indicator.

[0047] (4) Brain natriuretic peptide (BNP) and amino-terminal pro-brain natriuretic peptide (NT-proBNP): They have poor specificity for coronary heart disease, and elevated levels may be caused by other cardiovascular diseases (such as atrial fibrillation) or systemic metabolic disorders.

[0048] At present, the screening of clinical coronary heart disease (including ACS) mainly relies on biomarkers such as troponin (hsTNT, cTNI) and phosphokinase (CK-MB), which have the following technical problems: (1) Current markers tend to focus on specific physiological functions (such as myocardial injury, inflammatory response, and cardiac mechanical stress). (2) Some markers (such as troponin) are more suitable for the diagnosis of acute events, but their ability to assess the risk stratification of coronary heart disease is limited. Therefore, patients with clinical suspected ACS often undergo coronary angiography, which is an invasive examination with limited patient acceptance, high cost, and the risk of radiation exposure.

[0049] This application discovered and proposed for the first time the use of MPC1 specifically expressed in circulating exosomes as a diagnostic biomarker for coronary heart disease, providing a new solution for the detection of coronary heart disease.

[0050] Based on the discovery of the new diagnostic marker of the present application, the first aspect of the embodiments of the present application provides a use of a mitochondrial pyruvate transporter 1 and / or its detection reagent in the preparation of a coronary heart disease detection product.

[0051] This application also has the following advantages:

[0052] (1) Early diagnostic potential: Exosomal MPC1 can be manifested in the early stages of metabolic changes in coronary heart disease and can effectively improve the detection rate of early ACS patients.

[0053] (2) Higher sensitivity and specificity: MPC1 in circulating exosomes has higher specificity and can reflect the imbalance of mitochondrial energy metabolism in patients with coronary heart disease, rather than myocardial damage or systemic inflammatory response. Compared with other biomarker values ​​in plasma, exosome detection has higher sensitivity.

[0054] (3) Non-invasive detection: Compared with coronary angiography, exosomes are derived from blood samples, which is convenient for liquid biopsy and has great potential for clinical application.

[0055] In some embodiments of the present application, the sample detected by the coronary heart disease detection product includes body fluids of the circulatory system. Optionally, the sample detected by the coronary heart disease detection product includes blood, such as plasma. In some embodiments of the present application, the coronary heart disease detection product detects exosome mitochondrial pyruvate transporter 1 in the sample.

[0056] Circulating exosomes are small vesicles secreted by a variety of cells. The molecular components such as proteins and nucleic acids contained in them can reflect the physiological and pathological states of cells and have received extensive attention in the study of diseases such as coronary heart disease. In addition, mitochondrial dysfunction (such as respiratory chain damage, increased metabolic heterogeneity, etc.) is considered to be an important mechanism for the occurrence and development of coronary heart disease, especially the regulation of mitochondrial metabolic processes. In the mitochondrial energy metabolism pathway, MPC1 is a key transporter responsible for the entry of pyruvate into mitochondria. Its abnormal expression may affect the homeostasis of energy metabolism and thus participate in the pathological process of coronary heart disease. Therefore, the expression level and changes of MPC1 in circulating exosomes may represent the degree of occurrence and development of coronary heart disease and have the potential to be used as a biomarker. At present, the research on circulating exosomes is gradually deepening, and some progress has been made in the exploration of exosomal miRNAs, proteins and metabolites used for the exploration of coronary heart disease markers. The study of mitochondrial pyruvate transporter is still an emerging field, and there is still a large gap in its detection method in circulating exosomes and its clinical relevance in coronary heart disease. Therefore, this application fills the gap in this field and provides a more efficient diagnostic tool and potential therapeutic target for coronary heart disease.

[0057] It is understandable that the detection product of the present application may be a set product, further comprising one or more of an exosome extraction reagent and an exosome detection reagent.

[0058] The present application does not particularly limit the detection mechanism and detection method of the detection reagent, including but not limited to any of the following methods for detection: mass spectrometry, immunoassay, western blot analysis, radioimmunoassay, immunofluorescence, immunoprecipitation, equilibrium dialysis, immunodiffusion, electrochemiluminescence immunoassay, ELISA assay and immunopolymerase chain reaction.

[0059] The present application does not specifically limit the detection reagents. It is understandable that the types and components of the corresponding detection reagents may vary with different detection mechanisms and detection methods, including but not limited to: detection kits or chips.

[0060] In some embodiments of the present application, the coronary heart disease detected by the coronary heart disease detection product includes acute myocardial infarction. In some embodiments of the present application, the coronary heart disease detected by the coronary heart disease detection product includes myocardial necrosis. In some embodiments of the present application, the coronary heart disease detected by the coronary heart disease detection product includes acute coronary syndrome.

[0061] The second aspect of the embodiments of the present application provides a coronary heart disease detection product as defined above.

[0062] The embodiments of the present application will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present application and are not intended to limit the scope of the present application. The experimental methods for which specific conditions are not specified in the following examples are preferably referred to the guidance provided in the present application, and can also be based on the experimental manual or normal conditions in this area, can also be based on the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0063] In the following specific embodiments, the measured parameters of raw material components may have slight deviations within the range of weighing accuracy unless otherwise specified. For temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0064] 1. Establish a mouse acute myocardial infarction model, extract mouse plasma exosomes, identify and quantify MPC1 expression using ELISA

[0065] 1.1 Methods

[0066] (1) Construction of acute myocardial infarction model

[0067] C57BL / 6 mice aged 6-8 weeks were anesthetized with 5% isoflurane, fixed in supine position on the operating table, hair removed from the precordial area, and disinfected with 75% alcohol cotton balls. Under cold light, the tongue of the mouse was fixed with a tongue clip, and then a 1.5mm diameter endotracheal tube was used for oral endotracheal intubation, and the small animal ventilator was connected for assisted ventilation. Anesthesia was maintained with a 2% isoflurane concentration, and the ventilator parameters were adjusted to tidal volume 2.0mL / time and respiratory rate 120 times / min for assisted ventilation. Preoperative electrocardiogram was recorded. The skin was incised from the second intercostal space on the left side of the sternum with a sterile blade, and the subcutaneous tissue, pectoralis major and pectoralis minor were bluntly separated with hemostats, and the intercostal space was bluntly expanded to expose the heart. An 8-0 suture needle with thread was used to penetrate the 1 / 3 myocardial layer at 1-2mm below the lower edge of the left atrial appendage, and the left anterior descending branch of the coronary artery was ligated. At this time, the electrocardiogram showed ST segment elevation, and the myocardium turned white after ligation, the contractility was weakened, and the ST segment elevation > 1 / 2R wave) was maintained for 5 minutes, confirming that the model was successful. The sham operation group mice were opened to expose the heart, and the myocardium was penetrated by 8-0 metabolic suture needle without ligation as the control group.

[0068] (2) Extraction of mouse plasma exosomes

[0069] 2 mL of plasma samples were collected from two groups of mice (sham operation group and acute myocardial infarction group) by heart blood sampling method, 13 mL of phosphate buffered saline (PBS) was added, and centrifuged at 300 g for 10 min at 4 ° C. The supernatant was taken and then centrifuged at 12000 g for 40 min at 4 ° C. The supernatant was taken and filtered through a 0.22 μm filter membrane. Ultracentrifuged at 120000 g at 4 ° C for 1.5 h, and the supernatant was discarded. The precipitate was washed with 15 mL of PBS solution, and centrifuged at 100000 g at 4 ° C for 1 h to precipitate exosomes.

[0070] (3) Western Blot Identification of Exosome Markers

[0071] PBS was added to dissolve the above precipitated exosomes for exosome purity identification (positive marker: TSG101 / CD63 / CD9; negative marker: TBP / GM130). The specific steps are as follows:

[0072] a. Add the corresponding volume of 5× loading buffer according to the volume of PBS dissolved exosomes;

[0073] b. Protein denaturation at 99°C for 10 min;

[0074] c. Prepare SDS-PAGE gels of different concentrations according to the target molecular weight;

[0075] d. Add the processed exosome protein samples into the SDS-PAGE gel loading wells;

[0076] e. Complete electrophoresis and membrane transfer under constant voltage and constant current conditions respectively;

[0077] f. Block with 5% skim milk at room temperature for 1 hour;

[0078] g. Incubate the corresponding primary antibody according to different molecular weights, incubate overnight on a shaker at 4°C, and wash away the excess unbound primary antibody;

[0079] h. Incubate different secondary antibodies according to the source of primary antibody at room temperature for 1 hour, and wash away the excess unbound secondary antibody;

[0080] i. Incubate the membrane with luminescent solution to visualize protein bands.

[0081] (4) ELISA determination of exosomal MPC1 expression

[0082] PBS was added to dissolve the precipitated exosomes, and the expression of plasma exosome MPC 1 content was detected by ELISA experiment (kit: Cloud Clone SEQ978).

[0083] 1.2 Results

[0084] In this example, an acute myocardial infarction model was constructed in mice, plasma exosomes were extracted, positive exosome markers (TSG101 and CD9) and negative exosome markers (GM130) were identified, and ELISA was used to determine the expression of MPC1. Figure 1 shown.

[0085] Figure 1 middle:

[0086] Figure A, Western blot detection of TSG101, CD9, and GM130 protein expression in mouse plasma exosomes (Exo). Mouse heart tissue (Tissue) was used as a control.

[0087] Figure B, Plasma exosomes were extracted, and the exosome protein concentrations of mice in the sham operation group (Sham) and acute myocardial infarction (MI) group were detected by BCA method. *** P<0.01, N=6.

[0088] 2. Extract exosomes from patient plasma and identify the purity of exosome extraction

[0089] 2.1 Sample

[0090] (1) Inclusion criteria

[0091] a. Over 18 years old;

[0092] b. Patients hospitalized in the Department of Cardiovascular Medicine of our hospital (The Second Affiliated Hospital of Guangdong Medical University) who are scheduled to undergo emergency coronary intervention in accordance with the "Guidelines for the Management of Acute Coronary Syndrome 2023".

[0093] (2) Exclusion criteria

[0094] a.Age < 18 years old;

[0095] b. Severe heart valve disease;

[0096] c. Hemodynamic instability requires assistance from IABP or ECMO or other devices;

[0097] d. Previous history of HF or MI;

[0098] e. Acute cerebrovascular accident or combined with severe liver and kidney dysfunction;

[0099] f. Combined with severe infection, systemic inflammatory disease or autoimmune disease;

[0100] g. Patients with severe bleeding tendency or recent history of bleeding disorders;

[0101] h. Pregnant or breastfeeding women;

[0102] i. Those who refuse to sign the informed consent form or have participated in other clinical trials in the past three months.

[0103] (3) Sample size

[0104] A total of 66 plasma samples that met the above inclusion and exclusion criteria were collected from the Department of Cardiology, the Second Affiliated Hospital of Guangdong Medical University from November 1, 2024 to February 28, 2025.

[0105] (4) Non-interfering samples

[0106] 2.2 Methods

[0107] 4 mL of blood was collected from the clinical control group and the experimental group and placed in an EDTA anticoagulant tube. The tube was centrifuged at 3000 × g for 15 min at 4 °C and the supernatant was aspirated, i.e., plasma. Exosomes were extracted from plasma using the precipitation method (kit: Gisai E1001). PBS was added to dissolve the precipitated exosomes for exosome purity identification (positive markers: TSG101 / CD63 / CD9; negative markers: TBP / GM130). The specific steps are as follows:

[0108] a. Add the corresponding volume of 5× loading buffer according to the volume of PBS dissolved exosomes;

[0109] b. Protein denaturation at 99°C for 10 min;

[0110] c. Prepare SDS-PAGE gels of different concentrations according to the target molecular weight;

[0111] d. Add the processed exosome protein samples into the SDS-PAGE gel loading wells;

[0112] e. Complete electrophoresis and membrane transfer under constant voltage and constant current conditions respectively;

[0113] f. Block with 5% skim milk at room temperature for 1 hour;

[0114] g. Incubate the corresponding primary antibody according to different molecular weights, incubate overnight on a shaker at 4°C, and wash away the excess unbound primary antibody;

[0115] h. Incubate different secondary antibodies according to the source of primary antibody at room temperature for 1 hour, and wash away the excess unbound secondary antibody;

[0116] i. Incubate the membrane with luminescent solution to visualize protein bands.

[0117] 2.3 Results

[0118] Western blot was used to identify exosome positive markers (TSG101 and CD9) and negative markers (GM130). Figure 2 shown.

[0119] 3. Analysis

[0120] 3.1 Collect the number of blood vessels with coronary artery stenosis and the degree of vascular stenosis after angiography in patients hospitalized in the Department of Cardiology who plan to undergo coronary angiography.

[0121] 3.2 Collect laboratory-related test indicators such as NT-proBNP and hs-TNT when patients are admitted to the hospital.

[0122] 3.3 Take 50 μL of the exosomes isolated in 2.2 and detect the expression of plasma exosome MPC1 content by ELISA experiment (kit: Cloud Clone SEQ978).

[0123] 3.4 The correlation between the MPC1 content in plasma exosomes and the number of coronary artery stenosis vessels, the most severe vascular stenosis, NT-proBNP and hs-TNT was calculated and analyzed, and the correlation between the two variables was evaluated by the Pearson correlation coefficient.

[0124] 3.5 Through ROC curve analysis, the efficiency of effectively screening patients with multiple coronary artery stenosis and patients with single coronary artery or no stenosis was compared with the traditional indicators NT-ProBNP, hs-TNT and plasma exosomal MPC1 molecule content.

[0125] 3.6 Through ROC curve analysis, the efficiency of effectively screening patients with coronary artery stenosis greater than 75% and patients with less than 75% or no stenosis was compared with the traditional indicators NT-ProBNP, hs-TNT and plasma exosomal MPC1 molecule content.

[0126] Correlation and ROC curve analysis results are as follows Figure 3 shown.

[0127] Figure 3 middle:

[0128] Figure A: ELISA method was used to compare the expression of plasma exosomal MPC1 in patients with myocardial infarction (MI) and non-MI coronary heart disease (No-MI CVD). * P<0.05, 20 patients with myocardial infarction and 44 patients with non-myocardial infarction coronary heart disease.

[0129] Figure B, ELISA method was used to compare the expression of MPC1 protein in the plasma of patients with myocardial infarction (MI) and non-MI coronary heart disease (No-MI CVD).

[0130] Figure C, Correlation between the expression level of plasma exosomal MPC-1 and the amount of coronary artery stenosis in patients.

[0131] Figure D, correlation between the expression of plasma exosomal MPC-1 and the degree of coronary artery stenosis in patients.

[0132] In Figures C and D, 1-no coronary artery stenosis; 2-coronary artery stenosis ≤30%; 3-coronary artery stenosis >30% and ≤40%; 4-coronary artery stenosis >40% and ≤50%; 5-coronary artery stenosis >50% and ≤60%; 6-coronary artery stenosis >60% and ≤75%; 7-coronary artery stenosis greater than 75%.

[0133] Figure E shows the ROC curve reflecting the predictive value of plasma exosomal MPC-1 expression level in predicting the cumulative number of coronary artery stenosis (with the number of involved vessels ≥ 2 as the critical point). The predictive ROC curve of hs-TNT / NT-proBNP was used as the control group.

[0134] Figure F, ROC curve of the predictive value of plasma exosomal MPC-1 expression level in predicting the degree of coronary artery stenosis (with stenosis ≥ 75% as the critical point). The predictive ROC curve of hs-TNT / NT-proBNP was used as the control group.

[0135] The technical features of the above-mentioned implementation modes and examples can be combined in any appropriate manner. To make the description concise, not all possible combinations of the technical features in the above-mentioned implementation modes and examples 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.

[0136] The above-described embodiments only express several implementation methods of the present application, which is convenient for understanding the technical solution of the present application in detail, but it cannot be understood as a limitation on the scope of protection of the patent application. It should be pointed out that for ordinary technicians in this field, without departing from the concept of the present application, several deformations and improvements can be made, which all belong to the protection scope of the present application. In addition, it should be understood that after reading the above-mentioned teaching content of the present application, the technicians in this field can make various changes or modifications to the present application, and the equivalent forms obtained also fall within the protection scope of the present application. It should also be understood that the technical solutions obtained by the technicians in this field through logical analysis, reasoning or limited experiments on the basis of the technical solutions provided in the present application are all within the protection scope of the claims attached to the present application. Therefore, the protection scope of the patent of the present application shall be based on the content of the attached claims, and the description and drawings can be used to explain the content of the claims.

Claims

1. Application of mitochondrial pyruvate transporter 1 and / or its detection reagent in the preparation of coronary heart disease detection products.

2. The use according to claim 1, characterized in that: The samples tested by the coronary heart disease detection product include body fluids of the circulatory system.

3. The use according to claim 2, characterized in that: The samples tested by the coronary heart disease detection product include blood.

4. The use according to claim 3, characterized in that: The coronary heart disease detection product detects exosome mitochondrial pyruvate transporter 1 in a sample.

5. The use according to claim 4, characterized in that: The coronary heart disease detection product also includes one or more of an exosome extraction reagent and an exosome detection reagent.

6. The use according to any one of claims 1 to 5, characterized in that The detection reagent in the coronary heart disease detection product is detected by any of the following methods: mass spectrometry, immunoassay, western blot analysis, radioimmunoassay, immunofluorescence, immunoprecipitation, equilibrium dialysis, immunodiffusion, electrochemiluminescence immunoassay, ELISA assay and immunopolymerase chain reaction.

7. The use according to any one of claims 1 to 5, characterized in that The coronary heart disease detection product is a detection kit or a chip.

8. The use according to any one of claims 1 to 5, characterized in that The coronary heart disease detected by the coronary heart disease detection product includes acute myocardial infarction.

9. The use according to any one of claims 1 to 5, characterized in that The coronary heart disease detected by the coronary heart disease detection product includes acute coronary syndrome.

10. The coronary heart disease detection product as defined in any one of claims 1 to 9.

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