An lncRNA-HRAT and its applications

Abstract

The present invention belongs to the field of biomedical technology, and specifically relates to an lncRNA-HRAT and its application. The present invention simulates the in vivo ischemia / reperfusion process through an hypoxia / reoxygenation (H / R) model, and then screens for lncRNAs with large differences in expression levels before and after H / R treatment through high-throughput sequencing technology. Finally, the lncRNA-HRAT is screened out. The lncRNA-HRAT provided by the present invention can be used as a biomarker for diseases such as myocardial infarction and myocardial ischemia-reperfusion injury, with high sensitivity, capable of detecting myocardial infarction and myocardial ischemia-reperfusion injury diseases and having high diagnostic efficiency and good reliability, providing a new detection method for diagnosing myocardial infarction and myocardial ischemia-reperfusion injury diseases. The detection method is simple and easy to operate, and has good potential for translational medicine prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to the application of lncRNA-HRAT. Background Art

[0002] For patients with acute myocardial infarction, the most effective method to rescue ischemic myocardium is to rapidly restore the blood supply to the ischemic myocardium, that is, reperfusion. Currently, thrombolytic therapy or effective interventional treatment means such as percutaneous transluminal coronary angioplasty, coronary artery bypass grafting, and laser myocardial revascularization are often used. However, after the blood supply is restored again, the damage of tissues or organs may not be reduced but further aggravated within a subsequent period of time, manifested as arrhythmia, myocardial contractility and relaxation dysfunction, metabolic abnormalities, and changes in myocardial ultrastructure, that is, myocardial ischemia / reperfusion (I / R) injury.

[0003] At present, the main mechanisms of myocardial ischemia-reperfusion injury are as follows: reactive oxygen species (ROS) injury, calcium overload, energy metabolism disorder, inflammatory response, apoptosis, autophagy, etc. However, the mechanism of myocardial ischemia-reperfusion injury has not been fully elucidated. Therefore, it is of great significance to conduct in-depth research on myocardial ischemia-reperfusion injury to find new preventive measures and treatment targets.

[0004] Research shows that only less than 2% of the RNA in the human transcriptome can encode proteins, and the rest are mainly non-coding RNAs (ncRNAs), including ribosomal RNAs (rRNAs), transfer RNAs (tRNAs), microRNAs (miRNAs), long non-coding RNAs (lncRNAs), circular RNAs (circRNAs), and other small RNAs. Among them, lncRNAs are a class of functional RNA molecules with a transcript length exceeding 200 nt (nucleotide units), and they lack the ability to encode proteins. LncRNAs are usually longer, have an mRNA-like structure, are spliced, have a polyA tail and a promoter structure, and have dynamic expression and different splicing patterns during the differentiation process. However, compared with most mRNAs, they have fewer and longer exons, relatively low expression levels, poor primary sequence conservation, and high cell and tissue specificity. Research has confirmed that lncRNAs are involved in many important biological phenomena, such as X chromosome silencing, imprinted genomic loci, formation of chromosome conformation, and regulation of allosteric enzyme activity.

[0005] Aberrant expression of lncRNAs is closely associated with various diseases, such as cardiovascular diseases, neurological diseases, cancers, etc. Therefore, lncRNAs can serve as new targets for disease treatment. Recent studies have shown that lncRNAs are key components of the gene regulatory network controlling cardiovascular development, and aberrant expression of lncRNAs is closely related to the occurrence and development of heart diseases. Some studies have confirmed that there are some aberrantly expressed lncRNAs in the heart tissue of mice during early reperfusion after myocardial ischemia. Subsequent studies have further confirmed that lncRNAs play an important role in myocardial ischemia-reperfusion injury. For example, lncRNA-H19 can reduce cardiomyocyte apoptosis and myocardial ischemia-reperfusion injury by inhibiting the miR-877-3p-Bcl-2-mediated mitochondrial apoptosis pathway; inhibiting the expression of lncRNA-TUG1 can reduce its adsorption of miR-142-3p, thereby downregulating the expression of miR-142-3p target genes HMGB1 and Rac1, and ultimately improving myocardial injury caused by ischemia / reperfusion; inhibiting the expression of lncRNA-XIST can improve myocardial injury caused by ischemia / reperfusion by regulating the miR-133a / SOCS2 axis and inhibiting autophagy. However, the intervention of these lncRNA molecules partially improves myocardial ischemia-reperfusion injury, but whether there are other lncRNAs that regulate myocardial ischemia-reperfusion injury remains to be further studied.

[0006] Therefore, the present invention intends to screen out new lncRNAs with aberrant expression during the process of hypoxia / reoxygenation injury of cardiomyocytes by RNA sequencing, and explore their roles in myocardial ischemia-reperfusion injury, so as to provide new targets and theoretical basis for the treatment of myocardial ischemia-reperfusion injury. Summary of the Invention

[0007] To solve the technical problems existing in the prior art, the present invention provides an lncRNA-HRAT, which can be used as a biomarker for diseases such as myocardial infarction and myocardial ischemia-reperfusion injury, or can be used to prepare drugs for preventing / treating / diagnosing diseases such as myocardial infarction and myocardial ischemia-reperfusion injury.

[0008] The technical solution of the present invention is as follows.

[0009] In the first aspect, the present invention provides an lncRNA-HRAT, which contains the nucleotide sequence shown in SEQ ID NO:1 or SEQ ID NO:2.

[0010] In the second aspect, the present invention also provides a nucleic acid molecule encoding the above lncRNA-HRAT and a vector containing the above nucleic acid molecule.

[0011] In a third aspect, the present invention also provides a cell comprising the above-mentioned lncRNA-HRAT, or a nucleic acid molecule of the above-mentioned lncRNA-HRAT, or a vector comprising the above-mentioned.

[0012] In a fourth aspect, the present invention also provides the use of the above-mentioned lncRNA-HRAT as a biomarker for myocardial infarction or myocardial ischemia-reperfusion injury diseases, or its use in screening drugs for preventing and treating myocardial infarction or myocardial ischemia-reperfusion injury diseases.

[0013] In a fifth aspect, the present invention also provides a primer pair for detecting the above-mentioned lncRNA-HRAT, comprising an upstream primer and a downstream primer. The upstream primer: 5'-GGGTTTCATCGTCACGCAAG-3' (SEQ ID NO:3), the downstream primer: 5'-GGACCGTCTACCCCACACTA-3' (SEQ ID NO:4); or, the upstream primer: 5'-GTTATCTTTGTGGGTTCTTTAGCC-3' (SEQ ID NO:5); the downstream primer: 5'-GTTGATCTGTAGCCTCTCTGCC-3' (SEQ ID NO:6).

[0014] In a sixth aspect, the present invention also provides a diagnostic kit, which comprises the above-mentioned lncRNA-HRAT, or a nucleic acid molecule of the above-mentioned lncRNA-HRAT, or the above-mentioned vector, or the above-mentioned primer.

[0015] In a seventh aspect, the present invention also provides a method for detecting the above-mentioned lncRNA for non-disease diagnosis or treatment purposes, which is characterized by comprising the following steps:

[0016] (1) Extract the total RNA of the myocardial cells after H / R treatment to be detected, and reverse transcribe it into cDNA;

[0017] (2) Using the cDNA as a template, perform real-time fluorescence quantitative PCR detection with the above-mentioned primer pair;

[0018] (3) Calculate the expression level of the lncRNA-HRAT by the relative quantification method.

[0019] Furthermore, the reaction system for the real-time fluorescence quantitative PCR detection is a 20 μL PCR reaction system: 11 μL of 2× polymerase chain reaction premix, 1 μL of cDNA template, 1 μL of 10 μM upstream primer, 1 μL of 10 μM downstream primer, 7 μL of RNase-free water. Among them, the 2× polymerase chain reaction premix includes 2.5 U Taq DNA polymerase, 1.5 mmol / L MgCl2, 100 μmol / L dNTPs and 2.0 mmol / L SYBR Green I.

[0020] Furthermore, the PCR reaction conditions are as follows: pre-denaturation at 95°C for 30 seconds first; then denaturation at 95°C for 5 seconds, annealing and extension at 60°C for 34 seconds, for a total of 40 cycles.

[0021] The present invention has the following advantages:

[0022] The present invention screened out a new lncRNA-HRAT and verified its role in myocardial ischemia-reperfusion injury through in vivo and in vitro experiments. The results showed that in cardiomyocytes treated with H / R, overexpression of lncRNA-HRAT could significantly increase the expression of the apoptotic effector protein caspase-3, promoting H / R-induced cardiomyocyte apoptosis. In mice treated with I / R, knocking out lncRNA-HRAT could reduce the release of CK in serum after ischemia / reperfusion, reduce the myocardial infarction area, and improve cardiac function, indicating that upregulation of lncRNA-HRAT expression could exacerbate myocardial injury caused by ischemia / reperfusion. Therefore, lncRNA-HRAT may become a potential target for the treatment of myocardial ischemia-reperfusion injury. Description of the Drawings

[0023] Figure 1 : Expression changes of lncRNAs after H / R treatment, where, A. Heat map of differential expression of lncRNAs obtained by RNA-Seq; B. Candidate lncRNAs selected from the sequencing results; C. Expression of candidate lncRNAs detected by qRT-PCR. *P<0.05, **P<0.01, ns indicates no significant difference, (n = 5).

[0024] Figure 2 : Coding ability of lncRNA-HRAT calculated by Potential Calculator Online software.

[0025] Figure 3 : Subcellular localization of lncRNA-HRAT in cardiomyocytes (scale bar: 10 μm).

[0026] Figure 4 : Effect of overexpression of lncRNA-HRAT on caspase-3 expression, where, A. Expression level of lncRNA-HRAT in cardiomyocytes detected by qRT-PCR; B. Expression level of caspase-3 in cardiomyocytes detected by Western blot. *P<0.05, **P<0.01, (n = 3).

[0027] Figure 5:Effect of overexpressing lncRNA-HRAT on H / R-induced cell apoptosis, where, A. Flow cytometry results; B. Statistical analysis of flow cytometry results. *P<0.05, **P<0.01, (n = 3).

[0028] Figure 6 :Effect of I / R treatment on CK level in mouse serum and cardiac infarction area, where, A. Detecting the activity of CK in serum with CK kit; B. Detecting the cardiac infarction area of mice by TTC staining, normal myocardial tissue is red, infarcted myocardial tissue is white and statistical analysis of mouse myocardial infarction area. **P<0.01, (n = 5).

[0029] Figure 7 :Effect of I / R treatment on the expression of lncRNA-HRAT in cardiac tissue. **P<0.01, (n = 5).

[0030] Figure 8 :Construction process of HRAT-CKO mice, where, A. Gene construction strategy of HRAT-CKO mice; B. Line establishment process of HRAT-CKO mice.

[0031] Figure 9 :Genotype identification results of HRAT-CKO mice, where, A. Identification of Flox allele: Flox homozygote is 281bp, wild type is 211bp, Flox heterozygote has bands at both 281bp and 211bp; B. Identification of Cre allele: Cre is 304bp, wild type has no band; 0 is the blank control group, 1 - 7 are different HRAT-CKO mouse DNA sample groups.

[0032] Figure 10 :Expression level of lncRNA-HRAT in cardiac tissue of HRAT-CKO mice. **P<0.01, (n = 5).

[0033] Figure 11 :Effect of myocardial-specific knockout of lncRNA-HRAT on CK level in mouse serum. *P<0.05, **P<0.01, ns means no significant difference, (n = 5).

[0034] Figure 12 :Effect of myocardial-specific knockout of lncRNA-HRAT on cardiac infarction area of mice, where, A. Detecting the cardiac infarction area of mice by TTC staining, normal myocardial tissue is red, infarcted myocardial tissue is white; B. Statistical analysis of mouse myocardial infarction area. *P<0.05, (n = 5).

[0035] Figure 13:Effect of myocardial - specific knockout of lncRNA - HRAT on mouse heart function. Among them, A. Detection of mouse heart function by ultrasonic imaging system; B. Quantitative analysis of left ventricular ejection fraction (LVEF); C. Quantitative analysis of left ventricular fractional shortening (LVFS). *P < 0.05, **P < 0.01, ns indicates no significant difference, (n = 5). Detailed implementation manners

[0036] The following embodiments are only examples for better further understanding of the present invention, and do not limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior - art features falls within the protection scope of the present invention.

[0037] For those not specifying specific experimental steps or conditions in the embodiments, the operations or conditions of the conventional experimental steps described in the prior art in this field can be followed. For reagents or instruments not indicating the manufacturer, they are all conventional reagent products that can be obtained through commercial purchase.

[0038] Example 1 Screening of lncRNA - HRAT

[0039] To find lncRNAs that may play a role in myocardial ischemia - reperfusion injury, an hypoxia / reoxygenation (H / R) model was used to simulate the in - vivo ischemia / reperfusion process. Subsequently, 2 pairs of cell samples treated with H / R and the corresponding control groups were collected and sent to Guangzhou Ribobio Co., Ltd. for RNA - Seq sequencing analysis.

[0040] Taking the difference multiple between the H / R group and the control group > 2 (P < 0.05) as the screening criterion, a total of 1029 differentially expressed lncRNAs were screened out. Among them, 222 were up - regulated and 807 were down - regulated ( Figure 1 A). According to the following criteria: (1) lncRNAs with a difference multiple between the H / R group and the control group > 4.5; (2) not reported in the UCSC and Ensemble databases, we selected 7 lncRNAs ( Figure 1 B). The expression levels of these lncRNAs in H9C2 cardiomyocytes were detected by qRT - PCR to further verify the sequencing results. The results showed that compared with the control group, the changing trends of the two lncRNAs, TCONS_00029632 and TCONS_00078211, after H / R treatment were consistent with the RNA - sequencing results ( Figure 1 C). Among them, the change multiple of lncRNA TCONS_00029632 was the most significant after H / R treatment.

[0041] Therefore, lncRNA TCONS_00029632 may play a role in myocardial ischemia-reperfusion injury, so it was selected as the research object for subsequent studies. There is no relevant naming or report of lncRNA TCONS_00029632 in the prior art. According to its possible function, we named it lncRNA-HRAT (hypoxia / reoxygenation associated transprict, HRAT, hypoxia / reoxygenation-related transcript). The nucleotide sequence is shown in SEQ ID NO:1 or SEQ ID NO.2, where SEQ ID NO:1 is from Rattus norvegicus and SEQ ID NO:2 is from Mus musculus.

[0042] Example 2 Identification of lncRNA-HRAT

[0043] 2.1 Calculation of the coding ability of lncRNA-HRAT

[0044] To verify whether lncRNA-HRAT has coding ability, we used the PotentialCalculator Online software reported in the literature and input the lncRNA sequence for calculation. The results showed ( Figure 2 ): The coding ability values of the β-actin mRNA and Tp53 mRNA of the positive control were 3.70 and 8.25 respectively, while the coding ability value of lncRNA-HRAT was only -1.03303, indicating that it hardly has coding ability and belongs to non-coding RNA.

[0045] 2.2 LncRNA-HRAT is distributed in both the nucleus and cytoplasm

[0046] The different distribution positions of lncRNA in cells determine its mode of action. Therefore, it is very necessary to determine the subcellular localization of lncRNA-HRAT in cardiomyocytes for subsequent mechanism studies. We used RNA fluorescence in situ hybridization experiments to detect the localization of lncRNA-HRAT in cardiomyocytes. The results showed that: the internal reference gene 18S, as a positive control RNA, was mainly present in the cytoplasm, and lncRNA-HRAT was distributed in both the nucleus and cytoplasm ( Figure 3 ).

[0047] Example 3 LncRNA-HRAT promotes hypoxia / reoxygenation-induced cardiomyocyte injury in vitro

[0048] Cardiomyocytes can undergo apoptosis after hypoxia / reoxygenation treatment. Therefore, we regard cardiomyocyte apoptosis as a marker of cardiomyocyte injury. To verify the role of lncRNA-HART in myocardial ischemia-reperfusion injury, we first transfected lentivirus containing lncRNA-HRAT into H9c2 cells in vitro to increase the expression of lncRNA-HRAT in H9c2 cells, and then subjected H9c2 cells to H / R treatment. The expression of apoptotic effector protein caspase-3 was detected by western blot. The results showed that: compared with the negative control group, the expression level of lncRNA-HRAT in H9c2 cells increased significantly after lentivirus infection ( Figure 4 A). Compared with the normal control group, the expression level of caspase-3 increased significantly after H / R treatment of H9c2 cells, indicating that the H / R model was successfully established; compared with the negative control group, overexpression of lncRNA-HRAT significantly increased the expression level of caspase-3 ( Figure 4 B), indicating that lncRNA-HRAT can promote H / R-induced cardiomyocyte apoptosis.

[0049] To verify the regulatory effect of lncRNA-HRAT on cardiomyocyte apoptosis, we used flow cytometry to detect the change of apoptosis rate after overexpression of lncRNA-HRAT. The results showed ( Figure 5 ): compared with the normal control group, the apoptosis rate of H9c2 cells increased significantly after H / R treatment; compared with the negative control group, the apoptosis rate increased significantly after overexpression of lncRNA-HRAT, further confirming that lncRNA-HRAT can promote H / R-induced cardiomyocyte apoptosis, suggesting that lncRNA-HRAT can promote H / R-induced cardiomyocyte injury.

[0050] Example 4 Expression of LncRNA-HRAT in a Mouse Model of Myocardial Ischemia / Reperfusion Injury in Vivo

[0051] 4.1 Preparation of a Mouse Model of Myocardial Ischemia-Reperfusion Injury

[0052] After wild-type C57BL / 6 mice were subjected to ischemia for 45 min and reperfusion for 3 h, the serum of the modeled mice was taken, and the activity of CK in the serum was detected by an automatic biochemical analyzer; the hearts of the modeled mice were taken, and the infarct area of the heart was detected by TTC staining. The results showed that: compared with the sham operation (Sham) group, the activity of CK in the serum of mice increased significantly after I / R treatment ( Figure 6 A); the results of TTC staining showed that the infarct area of the I / R group was significantly larger than that of the Sham group ( Figure 6 B). The above results suggest that the mouse model of myocardial ischemia-reperfusion injury was successfully established.

[0053] 4.2 Upregulation of lncRNA-HRAT expression in the myocardial ischemia / reperfusion injury model

[0054] The in vitro experimental results of Example 3 showed that lncRNA-HRAT could promote H / R-induced cardiomyocyte injury. To explore the role of lncRNA-HRAT in in vivo myocardial ischemia / reperfusion (I / R) injury, we used qRT-PCR to detect the expression of lncRNA-HRAT in cardiac tissues after I / R treatment. The results showed that: compared with the Sham group, after I / R treatment, the expression of lncRNA-HRAT was significantly upregulated ( Figure 7 ), suggesting that lncRNA-HART might play a role in in vivo I / R injury.

[0055] Example 5 Construction of lncRNA-HRAT myocardial-specific knockout (HRAT-CKO) mice

[0056] 5.1 Genotype identification of HRAT-CKO mice

[0057] The principle of conditional gene knockout is to insert two LoxP sites at both ends of one or several important exons of the target gene to prepare mice containing two flox sites. Before the flox mice are crossed with mice expressing Cre recombinase, the gene is expressed normally; when the flox mice are crossed with mice expressing Cre enzyme specifically in tissues, the gene can be knocked out in specific tissues or cells, while the gene is expressed normally in other tissues or cells. Therefore, we can determine whether the gene in the mice is specifically knocked out by identifying whether the genotype of the mice is flox homozygous and carrying Cre enzyme.

[0058] HRAT-Flox homozygous (HRATF / F) mice and mice specifically expressing Cre enzyme in the myocardium (αMyHC-Cre) were provided by Cyagen Biosciences (Guangzhou) Co., Ltd. We bred lncRNA-HRAT myocardial-specific knockout (HRAT-CKO) mice by crossing the two, and the specific construction process of HRAT-CKO mice is as Figure 8 shown. We then collected the mouse tails for DNA extraction and amplification, and detected the genotype of the mice by agarose gel electrophoresis. The results showed that the Flox genotype of HRAT-CKO mice was homozygous and carried Cre enzyme ( Figure 9 ), suggesting that the knockout mice we constructed were HRAT-CKO mice.

[0059] 5.2 Expression of lncRNA-HRAT in the hearts of HRAT-CKO mice

[0060] To further verify the knockout effect of lncRNA-HRAT in the heart, we collected mouse hearts, extracted RNA, and used qRT-PCR to verify the expression of lncRNA-HRAT. The results showed that: compared with the mice in the HRATF / F group, the expression of lncRNA-HRAT in the hearts of CKO mice was significantly decreased ( Figure 10 ), indicating that the construction of lncRNA-HRAT heart-specific knockout mice was successful.

[0061] Example 6 Myocardial-specific knockout of lncRNA-HRAT can reduce myocardial ischemia-reperfusion injury

[0062] 6.1 Knockout of lncRNA-HRAT can reduce the level of CK in the serum of ischemia / reperfusion mice

[0063] To explore the role of lncRNA-HRAT in myocardial ischemia-reperfusion injury in vivo, after HRATF / F and HRAT CKO mice underwent ischemia for 45 min and reperfusion for 3 h, then a CK kit was used to detect the release of CK in the serum to reflect the degree of heart injury. The results showed ( Figure 11 ) that: compared with the Sham+HRATF / F group, the level of CK in the serum of the I / R+HRATF / F group increased significantly; compared with the mice in the I / R+HRATF / F group, the level of CK in the serum of the I / R+HRAT CKO group decreased significantly, indicating that myocardial-specific knockout of lncRNA-HRAT can reduce myocardial injury caused by ischemia / reperfusion.

[0064] 6.2 Knockout of lncRNA-HRAT can reduce the myocardial infarction area of ischemia / reperfusion mice

[0065] To verify the role of lncRNA-HRAT in myocardial ischemia-reperfusion injury in vivo, after HRATF / F and HRAT CKO mice underwent ischemia for 45 min and reperfusion for 24 h, we used TTC staining to detect the myocardial infarction area to reflect the degree of heart injury. The results showed ( Figure 12 ) that: compared with the mice in the HRATF / F group (43.57±5.29%), after knocking out lncRNA-HRAT, the myocardial infarction area of the mice decreased significantly (31.00±0.93%, p<0.05), indicating that knocking out lncRNA-HRAT can reduce myocardial infarction caused by myocardial ischemia / reperfusion, further confirming that myocardial-specific knockout of lncRNA-HRAT can reduce myocardial injury caused by ischemia / reperfusion.

[0066] 6.3 Knockout of lncRNA-HRAT can improve the heart function of ischemia / reperfusion mice

[0067] To further verify the role of lncRNA-HRAT in myocardial ischemia-reperfusion injury in vivo, 7 days after the ischemia-reperfusion experiment was performed on HRATF / F and HRATCKO mice, we used a small animal ultrasound imaging system to detect the left ventricular end-systolic diameter (LVEDS) and the left ventricular end-diastolic diameter (LVEDD), and then calculated the left ventricular ejection fraction (LVEF) and the left ventricular fractional shortening (LVFS) to evaluate the cardiac function of the mice. The results showed ( Figure 13 ): Compared with the Sham+HRATF / F group, the LVEF and LVFS in the I / R+HRATF / F group were significantly decreased; compared with the mice in the I / R+HRATF / F group, the LVEF and LVFS in the I / R+HRAT CKO group were significantly increased, suggesting that knocking out lncRNA-HRAT could improve the cardiac function abnormality caused by myocardial ischemia / reperfusion. The above results confirmed that myocardial-specific knockout of lncRNA-HRAT could reduce the myocardial injury caused by ischemia / reperfusion.

[0068] The above results indicated that the lncRNA-HRAT provided by the present invention could promote H / R-induced apoptosis / loss of cardiomyocytes, was up-regulated in the myocardial ischemia / reperfusion injury model, and myocardial-specific knockout of lncRNA-HRAT could reduce myocardial ischemia-reperfusion injury. Therefore, lncRNA-HRAT could be used as a biomarker for diseases such as myocardial infarction and myocardial ischemia-reperfusion injury, or for preparing drugs for preventing / treating / diagnosing diseases such as myocardial infarction and myocardial ischemia-reperfusion injury.

[0069] Obviously, the above embodiments are merely examples given for clear illustration and are not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation manners here. And the obvious changes or modifications derived therefrom are still within the protection scope of the present invention.

Claims

1. A lncRNA-HRAT, characterized in that, The nucleotide sequence of the lncRNA-HRAT is shown in SEQ ID NO:1 or SEQ ID NO:

2. SEQ ID NO:1 is derived from rats, and SEQ ID NO:2 is derived from mice.

2. A vector comprising the lncRNA-HRAT described in claim 1.

3. A cell comprising the vector described in claim 2.

4. Use of a reagent for detecting the lncRNA-HRAT described in claim 1 in the preparation of a detection reagent for diseases such as myocardial infarction or myocardial ischemia-reperfusion injury in rats or mice.

5. Use of the lncRNA-HRAT described in claim 1 in the screening of drugs for preventing and treating diseases such as myocardial infarction or myocardial ischemia-reperfusion injury in rats or mice.

6. A primer pair for detecting the lncRNA-HRAT described in claim 1, comprising an upstream primer and a downstream primer. Upstream primer: 5'-GGGTTTCATCGTCACGCAAG-3', downstream primer: 5'-GGACCGTCTACCCCACACTA-3'; or, upstream primer: 5'-GTTATCTTTGTGGGTTCTTTAGCC-3'; downstream primer: 5'-GTTGATCTGTAGCCTCTCTGCC-3'.

7. A diagnostic kit, characterized in that, The kit comprises the primer pair described in claim 6.

Images