CN114959003A

CN114959003A - Acute myocardial infarction marker and application thereof

Info

Publication number: CN114959003A
Application number: CN202111654193.9A
Authority: CN
Inventors: 魏慧卿; 赵蕾; 白龙
Original assignee: Second Hospital of Hebei Medical University
Current assignee: Second Hospital of Hebei Medical University
Priority date: 2021-12-30
Filing date: 2021-12-30
Publication date: 2022-08-30

Abstract

The invention relates to an acute myocardial infarction marker and application thereof. Compared with normal control, ELOVL6, SLAMF7 and TMEM88 as the markers of the invention show differential expression in patients with acute myocardial infarction, and the markers can be used alone or in combination to accurately and specifically diagnose the acute myocardial infarction.

Description

Acute myocardial infarction marker and application thereof

Technical Field

The invention belongs to the fields of biotechnology and medicine, and particularly relates to an acute myocardial infarction marker and application thereof.

Background

With the gradual modernization of society, the incidence of cardiovascular diseases is also increasing. Cardiovascular diseases are a serious health hazard due to their high morbidity and mortality. Acute Myocardial Infarction (AMI) is one of the most serious cardiovascular diseases, and is caused by severe and persistent Acute ischemic injury and necrosis of the heart muscle due to the sudden reduction or interruption of blood flow after the coronary artery is damaged or invaded by certain causes and formed into atherosclerotic plaques. AMI can be divided into two categories according to the electrocardiogram: ST-segment elevation myocardial infarction (STEMI) and Non-ST-segment elevation myocardial infarction (NSTEMI). STEMI is caused by total and long-term occlusion of the epicardial coronary arteries, whereas NSTEMI is usually caused by a transient occlusion due to stenosis of the coronary arteries or rupture of atherosclerotic coronary plaques followed by formation of a local thrombus. AMI patients may develop complications such as myocardial insufficiency, heart failure, arrhythmia, mechanical complications, and pericarditis, which are one of the main causes of death in AMI patients, and are more common in STEMI patients. With the significant increase in morbidity and mortality of AMI in recent years, and the trend toward ever-younger development, timely and accurate diagnosis is critical to controlling the occurrence and reducing mortality of AMI.

Disclosure of Invention

It is an object of the present invention to provide a method for accurately diagnosing AMI.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides an application of a reagent for measuring the expression quantity of the following biomarkers in a sample in preparing a tool for diagnosing acute myocardial infarction: ELOVL6, SLAMF7, and/or TMEM 88.

Further, the reagent includes a reagent for measuring the expression amount of the biomarker on the mRNA or protein level.

Further, the reagent for measuring the expression level of the biomarker on the mRNA level includes a primer, a probe specifically recognizing a nucleic acid sequence of the biomarker or a fragment of a complementary sequence thereof.

Further, said measuring the expression level of said biomarker at the mRNA level is achieved by a method selected from the group consisting of polymerase chain reaction, real-time fluorescent quantitation reverse transcription polymerase chain reaction, competitive polymerase chain reaction, nuclease protection assay, in situ hybridization, nucleic acid microarray, northern blot, or DNA chip.

Further, the reagent for measuring the expression level of the biomarker at the protein level includes an antibody, an antibody fragment, an aptamer, a high affinity multimer or a peptidomimetic that specifically recognizes the full length of the protein of the biomarker or a fragment thereof.

Further, the measuring of the expression level of the biomarker at the protein level is performed by a method selected from the group consisting of immunoblotting, enzyme-linked immunosorbent assay, radioimmunoassay, radioimmunodiffusion, immunoelectrophoresis, tissue immunostaining, immunoprecipitation analysis, complement fixation analysis, fluorescence-activated cell sorting, or protein microarray.

Further, the tool comprises a nucleic acid membrane strip, a preparation, a chip and a kit.

Further, the sample comprises blood and cells.

In another aspect of the present invention, there is provided a method of screening a candidate compound for the treatment of acute myocardial infarction, the method comprising:

(1) in the test group, administering a test compound to a subject to be tested, and detecting the level of the biomarker in a sample derived from said subject in the test group V1; in a control group, administering a blank control to the subject to be tested, and detecting the level of the biomarker in a sample derived from the subject in the control group, V2;

(2) comparing the level V1 and the level V2 detected in the previous step to determine whether the test compound is a candidate compound for treating acute myocardial infarction;

the biomarkers include ELOVL6, SLAMF7, and/or TMEM 88.

In another aspect, the invention provides the use of a biomarker comprising ELOVL6, SLAMF7 and/or TMEM88 in the screening of candidate compounds for the treatment of acute myocardial infarction.

Drawings

FIG. 1 is a boxplot of the relative expression levels of ELOVL 6;

FIG. 2 is a boxplot of the relative expression levels of SLAMF 7;

FIG. 3 is a box plot of the relative expression levels of TMEM 88;

FIG. 4 is a ROC plot of ELOVL6 in diagnosing acute myocardial infarction;

figure 5 is a ROC graph of SLAMF7 diagnosing acute myocardial infarction;

FIG. 6 is a ROC plot of TMEM88 diagnosing acute myocardial infarction;

FIG. 7 is a ROC plot of ELOVL6+ SLAMF7 in diagnosing acute myocardial infarction;

FIG. 8 is a ROC plot of SLAMF7+ TMEM88 in diagnosing acute myocardial infarction;

FIG. 9 is a ROC plot of ELOVL6+ TMEM88 diagnosing acute myocardial infarction;

FIG. 10 is a ROC plot of ELOVL6+ SLAMF7+ TMEM88 for the diagnosis of acute myocardial infarction.

Detailed Description

Hereinafter, the present invention will be described in detail by way of examples thereof with reference to the accompanying drawings. However, the following examples are provided as illustrations of the present invention, and when it is judged that a detailed description of a technique or a structure known to those of ordinary skill in the art to which the present invention pertains may unnecessarily obscure the gist of the present invention, a detailed description thereof may be omitted, and the present invention is not limited thereto. The present invention can be variously modified and applied within the scope of the following claims and equivalents to be explained thereby.

Also, the terms used in the present specification are terms used to appropriately express preferred embodiments of the present invention, and may vary according to the intention of a user or operator, the convention in the art to which the present invention pertains, and the like. Therefore, these terms should be defined based on the contents throughout the specification. In the present invention, the term "includes" or "including" a certain component in a certain portion is not intended to exclude another component but may include another component unless specifically stated to the contrary.

The term "and/or" as used herein in phrases such as "a and/or B" is intended to include both a and B; a or B; a (alone); and B (alone). Likewise, the term "and/or" as used in phrases such as "A, B and/or C" is intended to encompass each of the following embodiments: A. b and C; A. b or C; a or C; a or B; b or C; a and C; a and B; b and C; a (alone); b (alone); and C (alone).

Unless defined otherwise, 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 belongs. Any methods and materials similar or equivalent to those described herein can be used in the practice for testing the present invention, but the preferred materials and methods are described herein.

Reagent for measuring expression amount of biomarker in sample

In the present invention, the term "biomarker" refers to a biological molecule present in an individual at different concentrations that can be used to predict the disease state of the individual. Biomarkers can include, but are not limited to, nucleic acids, proteins, and variants and fragments thereof. A biomarker may be DNA comprising all or part of a nucleic acid sequence encoding the biomarker, or the complement of such a sequence. Biomarker nucleic acids useful in the present invention are considered to include DNA and RNA comprising all or part of any nucleic acid sequence of interest.

In a specific embodiment of the invention, the biomarker comprises ELOVL6, SLAMF7, and/or TMEM 88.

In the present invention, biomarkers such as ELOVL6(gene ID: 79071), SLAMF7(gene ID: 57823), TMEM88(gene ID: 92162), including gene and its encoded protein and its homologues, mutations, and isoforms. The term encompasses full-length, unprocessed biomarkers, as well as any form of biomarker that results from processing in a cell. The term encompasses naturally occurring variants (e.g., splice variants or allelic variants) of the biomarkers.

The term "sample" refers to a sample of bodily fluid, a sample of isolated cells or a sample from a tissue or organ. Body fluid samples may be obtained by well-known techniques and include samples of blood, urine, lymph, sputum, ascites, bronchial lavage or any other bodily secretion or derivative thereof. The tissue or organ sample may be obtained from any tissue or organ, for example, by tissue biopsy. Isolated cells may be obtained from a body fluid or tissue or organ by separation techniques, such as centrifugation or cell sorting, for example, a cell, tissue or organ sample may be obtained from such cells, tissue or organ that express or produce a biomarker. The sample can be frozen, fresh, fixed (e.g., formalin fixed), centrifuged, and/or embedded (e.g., paraffin embedded), and the like. Of course, prior to assessing the amount of label in the sample, the cell sample may be subjected to a variety of well-known post-collection preparation and storage techniques (e.g., nucleic acid and/or protein extraction, fixation, preservation, freezing, ultrafiltration, concentration, evaporation, centrifugation, etc.). Likewise, tissue biopsy samples may also be subjected to post-collection preparation and storage techniques, e.g., fixation. The sample may be collected before, during or after treatment. Samples may be taken from patients suspected of having or diagnosed with acute myocardial infarction and therefore may require treatment, or from normal individuals who are not suspected of having any condition. In the embodiment of the present invention, the sample is blood or cell.

The term "detecting" or "measuring" as used herein refers to quantifying the concentration of an object being detected or measured.

In the present invention, the term "primer" refers to a short nucleic acid sequence having a short free 3hydroxyl (free 3hydroxyl group) nucleic acid sequence capable of forming a base pair (base pair) with a complementary template (template), which serves as an origin for template strand replication. The primers can induce DNA synthesis in the presence of reagents for the polymerization reaction (i.e., DN polymerase or reverse transcriptase) and different 4 nucleoside triphosphates in the appropriate buffer and temperature.

In the present invention, the term "probe" refers to a nucleic acid fragment corresponding to several bases to several hundreds of bases capable of specifically binding to mRNA, for example, RNA or DNA, etc. Because of the labeling, the presence or absence of a specific mRNA can be confirmed. The probe can be produced in the form of an oligonucleotide (oligonucleotide) probe, a single-stranded dna (single stranded dna) probe, a double-stranded dna (double stranded dna) probe, an RNA probe, or the like.

The primer or probe of the present invention can be chemically synthesized by using a phosphoramidite solid phase support method or other known methods. Such nucleic acid sequences may be modified by a variety of means well known in the art. Non-limiting examples of such variations include methylation, encapsulation, substitution of more than one homolog of the natural nucleotide, and variations between nucleotides, for example, variations to uncharged linkers (e.g., methyl phosphonates, phosphotriesters, phosphoramidates, carbamates, etc.) or charged linkers (e.g., phosphorothioates, phosphorodithioates, etc.).

In the present invention, the term "antibody" is a term well known in the art and refers to a specific protein molecule directed against an antigenic site. For the purpose of the present invention, the antibody is an antibody that specifically binds to a protein encoded by the ELOVL6, SLAMF7, or TMEM88 gene, which is a marker of the present invention, and can be produced by a known method. Including partial peptides which can be made from the above proteins. The form of the antibody of the present invention is not particularly limited, and if a polyclonal antibody, a monoclonal antibody or any one having antigen binding property, a part thereof is also included in the antibody of the present invention, and all immunoglobulin antibodies are included. Furthermore, the antibody of the present invention also includes a specific antibody, for example, a humanized antibody.

Tool for diagnosing acute myocardial infarction

The tool for diagnosing acute myocardial infarction comprises a nucleic acid membrane strip, a preparation, a chip and a kit.

The chip of the invention comprises a gene chip and a protein chip; the gene chip comprises a solid phase carrier; and oligonucleotide probes immobilized on the solid support in an ordered manner, the oligonucleotide probes specifically corresponding to part or all of the sequences indicated by the biomarkers described above. The protein chip comprises a solid phase carrier and a specific antibody or ligand of the protein coded by the biomarker fixed on the solid phase carrier.

In the present invention, a nucleic acid membrane strip comprises a substrate and oligonucleotide probes immobilized on the substrate; the substrate may be any substrate suitable for immobilizing oligonucleotide probes, such as a nylon membrane, a nitrocellulose membrane, a polypropylene membrane, a glass plate, a silica gel wafer, a micro magnetic bead, or the like. Exemplary probes include PCR primers and gene-specific DNA oligonucleotide probes, such as microarray probes immobilized on a microarray substrate, quantitative nuclease protection test probes, probes attached to molecular barcodes, and probes immobilized on beads.

The present invention provides a kit which can be used to detect the levels of the biomarkers described above. The kit comprises a specific primer pair for amplifying the biomarker; a standard DNA template; and (3) PCR reaction liquid.

In a preferred embodiment, the specific primer pair comprises an upstream primer and a downstream primer.

The accuracy of the diagnosis of the disease is best described by the receiver-operating characteristics (ROC) of its subjects (Zweig, M.H. and Campbell, G., clinical chemistry 39(1993) 561-. The ROC plot is a plot of all sensitivity/specificity pairs obtained by varying the decision threshold over the entire range of data observed. The ROC plot shows the overlap between the two distributions by plotting the sensitivity versus 1-specificity over the complete decision threshold range. The y-axis is the sensitivity, or true positive rate defined as [ (number of true positive tests)/(number of true positive + false negative tests) ]. In the case of a disease or condition, this is also referred to as positive. It is computed only from the affected subgroups. The x-axis is the false positive rate, or 1-specificity [ defined as: (number of false positive results)/(number of true negative + false positive results) ]. This is an indicator of specificity and is calculated entirely from the unaffected subgroup. Because the true and false positive rates are calculated completely independently from the test results of the two different subgroups, the ROC plot is independent of prevalence in the sample. Each point in the ROC plot represents a sensitivity/specificity pair corresponding to a particular decision threshold. The ROC plot for a test with excellent discrimination (no overlap of the distributions of the two results) crosses the top left corner, where the true positive rate is 1.0 or 100% (perfect sensitivity) and the false positive rate is 0 (perfect specificity). A theoretical plot of the test without discriminatory power (the distribution of results for both clusters is the same) is a 45 ° diagonal line from the lower left to the upper right. Most of the graphs fall between these two extremes. Qualitatively, the closer the graph is to the upper left corner, the higher the overall accuracy of the test.

One purpose of quantifying the diagnostic accuracy of a laboratory test is to represent the performance of the laboratory test with a single number. The most common metric is the area under the curve (AUC) in the ROC plot. The area under the ROC curve is a measure of probability that enables correct identification of the condition. Conventionally, this area is typically greater than 0.5. The range of values was between 1.0 (test values for both populations were completely separated) and 0.5 (no significant difference in distribution between the two test values). This area depends not only on a certain part of the plot, such as the point closest to the slash or a sensitivity with a specificity of 90%, but on the entire plot. This quantitatively and descriptively expresses how close the ROC plot is to the perfect curve (area 1.0).

The concept and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention.

Example 1 screening for differentially expressed genes

First, research object

Blood samples were collected from 3 patients with acute myocardial infarction and 3 healthy controls.

Second, inclusion and exclusion criteria

1. Inclusion criteria

(1) Acute myocardial infarction patient

Inclusion criteria were: serum cardiac markers, mainly troponin, are elevated above at least 99% of the upper reference value, accompanied by CAG or CCTA confirmation of intracoronary thrombosis, one or more coronary artery main or branch stenosis, to an extent greater than 70%.

(2) Healthy control person

Inclusion criteria were: the electrocardiogram has no coronary heart disease characteristic, the serum myocardial enzyme marker is mainly normal troponin, and CAG or CCTA shows that no abnormal stenosis exists in the coronary artery lumen.

2. Exclusion criteria

(1) Patients suffering from other heart diseases, e.g. rheumatic heart disease, valvular heart disease or

Patients with congenital heart disease and the like;

(2) combined with diseases such as connective tissue disease, acute and chronic renal insufficiency, malignant tumor, or other diseases involving or associated with immune inflammation;

(3) combined with embolic diseases, such as disseminated intravascular coagulation, pulmonary embolism, lower limb arteriovenous embolism, mesenteric arteriovenous embolism and other blood vessel embolic diseases;

(4) patients taking anti-inflammatory drugs.

Third, the experimental procedure

1. Extraction of blood Total RNA

(1) And (3) homogenizing treatment: subject lml blood was collected using an EDTA-containing anticoagulation vacuum tube, following a 1: 3M1 TRIPURE LS Reagent was added and vortexed with a vortex mixer for 15-20s to lyse cells in the blood.

(2) Incubate at 15-30 ℃ for 5min to completely break down the nucleoprotein body.

(3) 0.6m1 chloroform was added, the sample tube cap was closed, shaken vigorously for 15s and incubated at room temperature for 3 min.

(4) After centrifugation at 12000rpm at 4 ℃ for 10min, the sample was separated into three layers, whereupon the RNA dissolved in the upper aqueous phase and the supernatant transferred to a fresh centrifuge tube.

(5) Adding equal volume of 75% ethanol, mixing by gentle inversion to give precipitate, and pouring the precipitate and liquid together into adsorption column RA sleeved with collecting tube.

(6) Centrifuging at 10000rpm at 4 deg.C for 45s repeatedly, and discarding waste liquid until all precipitate and solution pass through the column.

(7) Add 500. mu.l deproteinized liquid RE (Bioteke), centrifuge at 12000rpm for 45s, and discard the waste.

(8) Add 700. mu.l of Wash RW (Bioteke), centrifuge at 12000rpm for 60s, and discard the waste.

(9) Centrifuging at 4 deg.C and 12,000rpm for 2min to remove rinsing liquid as much as possible to prevent residual ethanol in the rinsing liquid from inhibiting downstream reaction.

(10) Taking out adsorption column RA, placing into centrifuge tube of RNase free, adding 80 μ L RNase free water (hot bath at 65-70 deg.C in advance), standing at room temperature for 2min, and centrifuging at 12000rpm for 1 min.

2. RNA quality detection

The concentration and purity of total RNA was determined using an ultraviolet spectrophotometer.

3. Library construction and transcriptome sequencing

(1) DNase digestion to remove DNA: DNA fragments existing in Total RNA samples are digested by DNase I, and reaction products are recovered by magnetic bead purification and finally dissolved in DEPC water.

(2) Removing rRNA: taking a digested Total RNA sample, removing rRNA by using a kit, carrying out Agilent 2100 detection after the rRNA is removed, and verifying the rRNA removal effect;

(3) RNA disruption: taking the sample in the previous step, adding a breaking Buffer, and placing the sample in a PCR instrument for thermal breaking to 130-;

(4) reverse transcription one-strand synthesis: adding a proper amount of primers into the broken sample, fully and uniformly mixing, reacting for a certain time at a proper temperature of a Thermomixer to open a secondary structure and combine with the primers, adding a one-chain synthesis reaction system Mix prepared in advance, and synthesizing one-chain cDNA on a PCR instrument according to a corresponding procedure;

(5) synthesis of reverse transcription duplex: preparing a double-chain synthesis reaction system, reacting on a Thermomixer at a proper temperature for a certain time to synthesize double-chain cDNA, and purifying and recovering reaction products by using magnetic beads. Purifying and recovering the product by using magnetic beads;

(6) and (3) repairing the tail end: preparing a terminal repair reaction system, reacting in a Thermomixer at a proper temperature for a certain time, and repairing the cohesive terminal of the cDNA double-chain obtained by reverse transcription under the action of enzyme. Purifying and recovering the end repairing product by using magnetic beads, and finally dissolving a sample in EB Solution;

(7) the cDNA ends were added with "A": preparing an A reaction system, reacting in a Thermomixer at a proper temperature for a certain time, and adding A basic groups to the 3' end of a product cDNA with repaired end under the action of enzyme;

(8) ligation of cdaadapter: preparing a joint connection reaction system, reacting in a Thermomixer at a proper temperature for a certain time, connecting a joint with the A base under the action of enzyme, and purifying and recovering a product by using magnetic beads.

(9) PCR reaction and product recovery: preparing a PCR reaction system, selecting a proper PCR reaction program, and amplifying the product obtained in the previous step. And (5) carrying out magnetic bead purification and recovery on the PCR product. The recovered product was dissolved in EB solution. Labeling and library preparation are completed.

(10) And (3) detecting the quality of the library: the size and concentration of fragments of the library were determined using an Agilent 2100 Bioanalyzer (Agilent DNA 1000 Reagents).

(11) Cyclization of PCR products: and (3) after the PCR product is denatured into a single chain, preparing a cyclization reaction system, fully mixing uniformly, reacting at a proper temperature for a certain time to obtain a single-chain cyclic product, and digesting the linear DNA molecules which are not cyclized to obtain the final library.

(12) And (3) machine sequencing: the single-stranded circular DNA molecule replicates through rolling circles to form a DNA Nanosphere (DNB) containing more than 200 copies. The obtained DNBs are added into the mesh pores on the chip by adopting a high-density DNA nano chip technology. The sequencing read length of 50bp/100bp is obtained by a sequencing-by-synthesis method.

4. Sequencing data quality control

Filtering the raw sequencing data to obtain high-quality sequencing data (clean data), comprising the following steps: removing an adapter sequence in reads; removing bases containing non-AGCT at the 5' end before shearing; pruning the ends of reads with lower sequencing quality (sequencing quality value less than Q20); removing reads with the N content of 10%; discarding small fragments with length less than 25bp after removing the adapter and mass pruning.

5. Alignment with reference genome

Sequencing data were aligned to the reference genome using hisat2 analytical software. The reference genome was from the Ensembl database.

6. Analysis of Gene expression levels

The expression level of the gene was calculated by aligning the number of sequences (clean reads) to the reference genomic region. The FPKM value of each gene/transcript in the sample was calculated using Stringtie according to the alignment of Hisat2 software, and this value was used as the expression level of the gene/transcript in the sample.

7. Differential mRNA expression analysis

The expression difference of mRNA of the control group and the disease group is compared by using DESeq2, and the difference analysis steps are as follows: firstly, standardizing (normalization) the original read count, mainly correcting the sequencing depth; carrying out hypothesis test probability (P-value) calculation through a statistical model, carrying out multiple hypothesis test correction (BH) to obtain a padj value (false discovery rate), wherein the screening standard of the differential expression genes is as follows: pvalue < 0.01.

Third, experimental results

Compared with a healthy control group, the blood expression of ELOVL6 and SLAMF7 in the acute myocardial infarction patient group is down-regulated, and the expression of TMEM88 is up-regulated.

Example 2 validation of differentially expressed genes in Large samples

Downloading GSE66360 data (the data set comprises circulating endothelial cell samples of 49 acute myocardial infarction patients and 50 healthy people) from a GEO database, annotating the probes on the genes by using GPL570 annotation documents, averaging the probes corresponding to a plurality of genes, carrying out differential analysis on the probes by using an R language limma package to obtain 1371 differentially expressed genes, wherein the screening standard is as follows: pvale < 0.001. The expression of the differentially expressed genes according to the present invention is shown in Table 1 and FIGS. 1 to 3.

TABLE 1 expression of biomarkers in patients with acute myocardial infarction

The invention uses R language pROC to carry out ROC diagnosis analysis on the gene so as to verify the diagnosis value of the gene. The results are shown in Table 2 and FIGS. 4 to 10.

TABLE 2 biomarker/biomarker combinations diagnostic potency

Biomarker/biomarker combinations	AUC
		ELOVL6	0.729
SLAMF7	0.714
		TMEM88	0.722
ELOVL6+SLAMF7	0.784
		SLAMF7+TMEM88	0.805
ELOVL6+TMEM88	0.793
		ELOVL6+SLAMF7+TMEM88	0.850

The experimental results demonstrate that the biomarker combinations have better diagnostic efficacy compared to the individual biomarkers. The results prove that the biomarker can be used for diagnosing acute myocardial infarction.

The preferred embodiments of the present application have been described in detail with reference to the accompanying drawings, however, the present application is not limited to the details of the above embodiments, and various simple modifications can be made to the technical solution of the present application within the technical idea of the present application, and these simple modifications are all within the protection scope of the present application. It should be noted that, in the foregoing embodiments, various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described in the present application. In addition, any combination of the various embodiments of the present application is also possible, and the same should be considered as disclosed in the present application as long as it does not depart from the idea of the present application.

Claims

1. Use of an agent for measuring the expression level of the following biomarkers in a sample for the preparation of a tool for diagnosing acute myocardial infarction: ELOVL6, SLAMF7, and/or TMEM 88.

2. The use of claim 1, wherein said reagents comprise reagents for measuring the expression of said biomarkers at the mRNA or protein level.

3. The use according to claim 2, wherein the reagent for measuring the expression level of the biomarker on the mRNA level comprises primers, probes specifically recognizing the nucleic acid sequence of the biomarker or a fragment of the complementary sequence thereof.

4. The use according to claim 3, wherein said measuring the expression level of said biomarker at the mRNA level is achieved by a method selected from the group consisting of polymerase chain reaction, real-time fluorescent quantitative reverse transcription polymerase chain reaction, competitive polymerase chain reaction, nuclease protection assay, in situ hybridization, nucleic acid microarray, northern blot, or DNA chip.

5. The use according to claim 2, wherein the reagent for measuring the expression level of the biomarker on the protein level comprises an antibody, an antibody fragment, an aptamer, a high affinity multimer or a peptidomimetic that specifically recognizes the full length of the protein of the biomarker or a fragment thereof.

6. The use according to claim 5, wherein said measuring the expression level of said biomarker at the protein level is performed by a method selected from the group consisting of immunoblotting, enzyme-linked immunosorbent assay, radioimmunoassay, radioimmunodiffusion, immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, fluorescence activated cell sorting, and protein microarray.

7. The use according to claim 1, wherein said means comprises a nucleic acid membrane strip, a preparation, a chip, a kit.

8. The use of claim 1, wherein said sample comprises blood, cells.

9. A method of screening a candidate compound for treatment of acute myocardial infarction, said method comprising:

(2) comparing the level V1 and the level V2 obtained from the previous step test to determine whether the test compound is a candidate compound for treating acute myocardial infarction;

the biomarkers include ELOVL6, SLAMF7, and/or TMEM 88.

10. Use of a biomarker for screening candidate compounds for the treatment of acute myocardial infarction, wherein the biomarker comprises ELOVL6, SLAMF7 and/or TMEM 88.