Application of lncRNA-AU020206 in preparation of medicine for treating cerebral arterial thrombosis

Abstract

The invention belongs to the technical field of gene drugs, and particularly relates to application of lncRNA-AU020206 in preparation of drugs for treating cerebral arterial thrombosis, the key effect of the lncRNA-AU020206 in occurrence and development of the cerebral arterial thrombosis is disclosed for the first time, and it is clear that the lncRNA-AU020206 activates ferroptosis signal channels through negative regulation of SLC7A11 expression, so that cerebral ischemia reperfusion injury is aggravated. In-vivo and in-vitro experiments prove that the levels of MDA, ROS and Fe < 2 + > can be remarkably reduced by silencing the lncRNA-AU020206, the expression of GPX4 and SLC7A11 can be up-regulated, the cerebral infarction volume and neurological dysfunction can be reduced, and cerebral edema and apoptosis can be improved. Therefore, the lncRNA-AU020206 not only can be used as a new target for treating the cerebral arterial thrombosis, but also can be used for developing a related diagnostic kit, and has remarkable clinical application prospect and market value.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of gene drugs, and particularly relates to application of lncRNA-AU020206 in preparation of a drug for treating ischemic stroke. BACKGROUND

[0002] Ischemic stroke is an acute cerebrovascular disease with high morbidity and high disability rate. Although modern medicine has made certain progress in blood vessel recanalization, such as intravenous thrombolysis and arterial thrombectomy, the ischemia / reperfusion process can induce strong oxidative stress and inflammatory response, and then induce various programmed cell death forms including ferroptosis, resulting in further damage to brain tissue. At present, there is still a lack of drug targets and intervention means that can effectively intervene in this process in clinical practice, especially a lack of effective drugs for secondary damage after reperfusion. Long-chain non-coding RNA (lncRNA) as an important regulatory molecule plays a key role in various diseases, but its functional mechanism in ischemic stroke is not clear.

[0003] Existing studies have shown that some lncRNAs such as lncRNA-AU020206 are up-regulated in a cerebral ischemia-reperfusion model, and may be involved in regulating cell death processes such as ferroptosis. However, its specific mechanism of action, interaction with YTHDC2 and the effect on SLC7A11 mRNA stability have not been systematically reported, and there is no literature reporting that the regulation pathway can be used as a new target for treating ischemic stroke. There is a lack of technical solutions for using it as a treatment target.

[0004] Therefore, it is urgent to develop a new treatment method with lncRNA-AU020206 and its downstream pathway-mediated ferroptosis pathway as the core of intervention to inhibit ferroptosis after stroke, protect neurons and improve prognosis. SUMMARY

[0005] In view of the above problems, the application of lncRNA-AU020206 in the preparation of a drug for treating ischemic stroke is provided. Through systematic research, it is first revealed that lncRNA-AU020206 is up-regulated in an ischemic stroke model, and mediates the ferroptosis pathway through regulating the YTHDC2 / SLC7A11 axis, and then a new technical solution for developing a drug for treating ischemic stroke by taking it as a target is proposed, filling the gap in the prior art.

[0006] To achieve the above purpose, the technical solution adopted by the present application is:

[0007] The present application provides application of lncRNA-AU020206 as a target for treating stroke.

[0008] The application also provides application of an inhibitor for inhibiting functional expression of lncRNA-AU020206 in preparation of a drug for treating a stroke.

[0009] The nucleic acid sequence of the lncRNA-AU020206 is shown as SEQ ID NO. 1.

[0010] Further, in the application, the inhibitor is shRNA for targeting and silencing expression of lncRNA-AU020206, and the nucleic acid sequence of the shRNA is shown as SEQ ID NO. 2.

[0011] The application also provides a pharmaceutical composition containing an effective amount of the lncRNA-AU020206 inhibitor and a pharmaceutically acceptable carrier, for treating an ischemic stroke.

[0012] Further, the carrier is one or more of chitosan, cholesterol, nanoparticles and liposomes.

[0013] Further, in the pharmaceutical composition, the dosage form of the pharmaceutical composition is oral preparation or injection preparation.

[0014] The application also provides application of a reagent for detecting expression level of lncRNA-AU020206 in vitro in preparation of a diagnostic kit for an ischemic stroke.

[0015] Further, in the application, the reagent includes specific primers or probes for lncRNA-AU020206.

[0016] By adopting the technical scheme, the application has the following beneficial effects:

[0017] 1. The application first discloses a key role of lncRNA-AU020206 in occurrence and development of an ischemic stroke. Through experimental research on a mouse focal cerebral ischemia-reperfusion (MCAO / R) model and an in vitro neuron hypoxia-reperfusion (OGD / R) model, it is found that lncRNA-AU020206 is significantly up-regulated in cerebral ischemia-reperfusion injury, and regulates mRNA stability of a ferroptosis key gene SLC7A11 by combining with m6A recognition protein YTHDC2, thereby affecting a neuron ferroptosis process. Silencing lncRNA-AU020206 can significantly reduce cerebral infarction volume, alleviate cerebral edema, improve neural function defects, and reduce MDA, ROS and Fe 2+Iron death related indicators. Therefore, lncRNA-AU020206 can not only be used as a new target for treating ischemic stroke, but also provides an experimental basis for its application in the preparation of oral or injection treatment drugs, and can be used for the development of related diagnostic kits, which has a significant clinical application prospect and market value.

[0018] 2. Based on the above mechanism, the present application further provides the use of an inhibitor (such as siRNA or shRNA) targeting lncRNA-AU020206 in the preparation of a drug for treating ischemic stroke. These inhibitors have high specificity and good biocompatibility, and can be delivered efficiently through various administration routes (such as intravenous injection, local administration, etc.), and have a good clinical application prospect. The implementation of the present application will provide a new idea for the precision treatment of ischemic stroke, and has significant scientific research value and potential market application potential. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 Figure 1 is a diagram of neurological score after MCAO / R of the present application using mNSS.

[0020] Figure 2 Figure 2 is a diagram of TTC-stained brain tissue sections and quantitative analysis of mouse brain infarction volume of the present application.

[0021] Figure 3 Figure 3 is a diagram of quantitative analysis of brain edema of the present application.

[0022] Figure 4 Figure 4 is a diagram of quantitative analysis of MDA, ROS and Fe 2+ content of the present application.

[0023] Figure 5 Figure 5 is a diagram of representative images of HE staining to detect ischemic penumbra cell pathology of the present application.

[0024] Figure 6 Figure 6 is a diagram of TUNEL staining to analyze ischemic penumbra cell apoptosis of the present application.

[0025] Figure 7 Figure 7 is a diagram of mRNA level detection of iron death related genes of the present application.

[0026] Figure 8 Figure 8 is a diagram of protein level detection of iron death related genes of the present application. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical scheme and advantages of the present application clearer, the present application is further described in detail below in combination with examples and drawings. It should be understood that the specific examples described herein are only used to explain the present application and do not limit the present application.

[0028] Example 1:

[0029] The embodiment of the present application provides a lncRNA-AU020206, the sequence cDNA of which is SEQ ID NO. 1.

[0030] The expression of the lncRNA-AU020206 is detected and the sequence thereof is obtained, and the specific process is as follows:

[0031] 1. Preparation of a focal cerebral ischemia-reperfusion animal model

[0032] The middle cerebral artery occlusion (MCAO) model of a mouse is prepared by referring to the modified Longa thread embolization method. 7-8-week-old C57BL / 6 male mice weighing 18-21 g are raised in a clean environment. First, the skin of the neck of the mouse is cleaned of hair with a hair clipper to expose the complete skin tissue, and then the skin is carefully cut with surgical scissors to make a 1.5-cm-long longitudinal incision in the middle of the neck. A surgical microscope is turned on, and the tissue around the neck of the mouse in the field of view is carefully pried open with a thicker forceps under the microscope to avoid mechanical damage. At this time, the tracheal muscle below is separated to expose the common carotid artery (CCA), and the muscle and fascia adhering to the CCA are further stripped with a microforceps. The bifurcation of the CCA can be seen upward, and the internal carotid artery (ICA) and external carotid artery (ECA) can be seen further forward. At this time, the CCA near the heart end and the ECA near the bifurcation are ligated. Since the pterygopalatine artery (PPA) can be artificially avoided from being mistakenly entered by the thread embolus, the PPA is not ligated. Then, a small opening is cut on the blood vessel with a microscissors near the ligation site of the CCA. A nylon thread embolus 0.18 mm coated with silicone at the head end is inserted into the CCA through the small opening. In order to avoid massive bleeding of the artery when the thread embolus is inserted, a slipknot is tied at the end of the blood vessel before the thread embolus is inserted, and then the slipknot is loosened and gently pushed to make the thread embolus advance along the internal carotid artery. After passing through the intersection of the ECA and the ICA, the thread embolus continues to advance along the internal carotid artery until it reaches the middle cerebral artery (MCA) of the mouse. When a slight resistance is felt and the thread embolus cannot be inserted further, the insertion is stopped. The distance from the bifurcation to the MCA is about 12-14 mm, which basically blocks the blood flow to the brain provided by the middle cerebral artery of the mouse. After 60 minutes of ischemia, the thread embolus is slowly pulled out, the external carotid artery is re-ligated, and then the silk thread at the common carotid artery is loosened to restore the blood flow of the right carotid artery. The incision is sutured, and then the mouse is placed in a cage after being disinfected with iodophor. The mouse is raised at room temperature of 25℃. The focal cerebral ischemia-reperfusion model is completed. The local cerebral blood flow is monitored by a laser Doppler flowmeter during the whole process to ensure the success of the model preparation. The body temperature of the mouse is maintained at 36.5℃±1℃ by using a body temperature instrument from the beginning of the operation to the awakening of the animal. The animals in the sham operation group are only exposed to the bifurcation of the internal and external carotid arteries after being anesthetized, without inserting the thread embolus.

[0033] 2. Experimental grouping and drug administration

[0034] C57BL / 6 male mice were randomly divided into 4 groups after weighing: sham operation group (Sham), cerebral ischemia model group (Vehicle) and intervention negative control group (shRNA-NC), intervention treatment group of lncRNA-AU020206 (shRNA-AU020206).

[0035] The sequence of shRNA is shown in SEQ ID NO. 2.

[0036] The sequence of SEQ ID NO. 2 is: 5'-GAAGCAAGCTTTATTTACATACTTCAAGAGAGTATGTAAATAAAGCTTGCTTTTTTTT-3';

[0037] Method of administration: according to the weight of mice, intraperitoneal injection of 10% chloral hydrate (4.0 ml / kg) anesthesia, prone position fixation, so that the skull remains horizontal and central. Shaving, iodophor disinfection, exposure of the skull, dip with appropriate amount of H2O2 wipe the skull, exposure of the fontanel point. According to the mouse brain atlas, the left ventricle coordinates are: anteroposterior direction = 0.8 mm, medial direction = ± 1.4 mm, dorsal abdominal direction = 3.5 mm relative to bregma. The plasmid containing shRNA-AU020206 or shRNA-NC was slowly injected into each lateral ventricle (3 μL per hemisphere, injection rate 0.15 μL / min, injection time more than 15 minutes), and the needle was left for 5 min after complete injection. After slowly lifting the micro-syringe needle, the bone wax was used to close the drill hole. The mouse was taken off from the stereotaxic instrument, and the wound was sutured, iodophor disinfection, and then put back to the feeding cage.

[0038] 3. Neurological function score

[0039] The mice were given 3 days of adaptation training before the modified neurological severity score (mNSS) was performed to make them familiar with the test room and a 1 m long, 3 cm wide wooden beam. The test was usually completed by the same operator who was unaware of the grouping to reduce subjective errors. The total score was 18 points, with 1 point for failure and 0 points for success, including four modules: ① motor coordination: tail lift observation of the contralateral forelimb flexion, free movement, and whether it continued to turn to one side or dragged gait, a total of 6 points; ② sensory function: use a cotton swab to brush the antennae and gesture to quickly approach the eyes, each 1 point for lack of avoidance reaction, a total of 2 points; ③ brainstem reflex: light touch cornea and near ear knock table, each 1 point for no blinking or jumping, a total of 2 points; ④ balance and coordination: the animal was placed in the center of the wooden beam and recorded 6 performances, 0 points for no falling, and the more times it fell, the higher the score, up to 6 points; if the laboratory needs, 2 points of supplementary tests such as hanging test or inclined plate retention can be added, but the total score still maintains 18 points. After completing each test, the scores were added up, with 0-4 points for mild, 5-10 points for moderate, and ≥11 points for severe functional impairment. In animal experiments, animals with excessive bleeding during surgery, abnormal respiration after surgery, premature death, and subarachnoid hemorrhage found at the time of sacrifice were discarded. The experimental results are shown in Figure 1 As shown in FIG. 8, the neurological motor score of the sh-AU020206 intervention treatment group was significantly lower than that of the model group, indicating that silencing lncRNA-AU020206 can significantly improve the neurological motor dysfunction of mice after cerebral ischemia-reperfusion.

[0040] 4. Measurement of cerebral infarction volume

[0041] The mouse brain was removed quickly 24 h after ischemia-reperfusion, the olfactory bulb, cerebellum and low brainstem were removed, and the mouse brain was frozen at -20°C for 20 min. Then the mouse brain was cut into 5 pieces from the frontal pole 3 mm, and the coronal sections were cut from front to back. The brain slices were incubated in 1% TTC at 37°C for 10 min, and the brain slices were occasionally turned to ensure uniform staining. After TTC staining, normal tissue was red and infarcted tissue was white. The mouse brain slices were scanned using Image J software and the infarction volume was calculated as a percentage of the contralateral brain volume. As shown in FIG. 9, compared with the model group and the negative control group, the cerebral infarction volume of the lncRNA-AU020206 silencing treatment group was significantly reduced. Figure 2

[0042] 5. Measurement of brain edema

[0043] ​After euthanizing the experimental mice, the skull was quickly opened, and both cerebral hemispheres were completely removed within 30 seconds (the cerebellum and brainstem were removed if necessary to ensure regional consistency). Excess blood was removed by gently touching the surface with fibrous filter paper, but the tissue was not squeezed. The brain tissue was immediately placed into a pre-calibrated and numbered aluminum weighing bottle, and the bottle was tightly sealed. The initial wet weight (Wwet) was recorded on a balance, accurate to 0.1 mg. The weighing bottle was then placed in a constant temperature oven at 100-105℃ for continuous drying for 24 hours. To ensure complete dehydration, the bottle cap could be slightly opened for 5 seconds after 12 hours to release moisture before continuing drying. After drying, the bottle was quickly transferred to a desiccator to cool for 30 minutes to prevent reabsorption of moisture from the air. The final dry weight (W) was then measured again. dry If the difference between the two weighings is less than 0.2 mg, it is considered that the drying is sufficient; otherwise, continue drying for 2 hours and weigh again. The final dry-to-wet ratio is calculated as W. wet / W dry This can also be expressed as brain water content (%) = W wet –W dry / W wet ×100. See attached. Figure 3 As shown, compared with the model group and the negative control group, the sh-AU020206 treatment group showed a significant reduction in cerebral edema. This result indicates that silencing lncRNA-AU020206 can significantly improve cerebral edema in mice after cerebral ischemia-reperfusion.

[0044] 6. Detection of MDA, ROS and Fe2+ content

[0045] After euthanasia of experimental mice, the target brain region (e.g., ischemic hemisphere cortex) was rapidly isolated within 30 seconds, homogenized with pre-cooled PBS + 0.1 mM EDTA in an ice bath, centrifuged at 12000 × g for 10 min at 4℃, and the supernatant was collected. Protein was quantified using the BCA method, and all indicators were normalized to mg protein. Subsequently, three indicators related to iron oxide death were detected sequentially: ① 100 μL of supernatant was mixed with TCA / SDS and TBA reaction solution, then boiled in a 95℃ water bath for 15 min, cooled in an ice bath, and centrifuged. The supernatant was then measured at 532 nm in a 96-well plate, and the lipid peroxidation end product MDA (U / mL) was calculated according to the MDA standard curve; ② 50 μL of supernatant was added with an equal volume of 10 μM DCFH-DA, incubated at 37℃ in the dark for 30 min, washed with PBS, and centrifuged twice. 100 μL of each well was measured at 488 / 525 nm in a black 96-well plate, and the results were expressed as ROS activity (U / mL); ③ 100 μL of supernatant was added sequentially with iron reducing agent and Ferrozine binding reagent, reacted at room temperature in the dark for 15 min, and measured at 593 nm. The results were calculated according to the Fe... 2+Standard curve was used to calculate the free ferrous content (pmol / L) and the whole process avoided the use of EDTA-containing buffer. All operations must be completed at 4℃ in the dark conditions, technical triplicate and biological repeat of no less than six animals were used to ensure that the colorimetric reading was completed within 30 min of the mixed reaction. As shown in Fig. 1, compared with the model group and the negative control group, the MDA, ROS and Fe Figure 4 content of the sh-AU020206 treatment group was significantly lower than that of the model group, indicating that silencing lncRNA-AU020206 can significantly improve the ferroptosis of mice after cerebral ischemia-reperfusion. 2+

[0046] 7. HE staining

[0047] The paraffin-embedded coronal brain sections (4 pm) were deparaffmized in xylene (2 x 5 min) and rehydrated in graded ethanol (100%, 95%, 80% and 70%) in distilled water; 2 min each). Then, the slides were immersed in hematoxylin solution for 5 min, rinsed in tap water for 2 min, and stained in PBS at pH 7.4 for 1 min. After a second brief rinse, the sections were counterstained with eosin for 90 s, washed in distilled water, and dehydrated with increasing concentrations of alcohol (95% and 100%); 2 min each). Finally, the brain tissue specimens were cleared in xylene (2 x 3 min) and covered with neutral resin. Morphological changes were observed under a light microscope (x200). As shown in Fig. 3, compared with the sham operation group, the number of cells in the cerebral cortex of the model group and the negative control group was significantly reduced, and the degree of karyopyknosis and vacuolization was severe; after treatment with sh-AU020206, the number of cells in the cortex of mice was significantly increased compared with the model group, and the cell morphology also recovered. Figure 5

[0048] 8. TUNEL staining

[0049] Neuronal apoptosis was detected by TUNEL (terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling) assay. Cells or brain tissue samples were fixed on slides with ice-cold 4% glutaraldehyde for 10 min, followed by permeabilization with 0.1% (v / v) Triton X-100 (Sigma) for 5 min. Slides were incubated with TUNEL incubation mixture for 1.5 h at 37℃ in a humid dark environment. Nuclei were then stained with 4,6-diamidino-2'-phenylindole hydrochloride (DAPI). TUNEL-positive staining was observed and photographed using a fluorescence microscope. The apoptosis index refers to the percentage of TUNEL-positive cells (positive cells / 100% total cells). As shown in Fig. 4, compared with the sham operation group, the number of cells in the cerebral cortex of the model group and the negative control group was significantly reduced, and the degree of karyopyknosis and vacuolization was severe; after treatment with sh-AU020206, the number of cells in the cortex of mice was significantly increased compared with the model group, and the cell morphology also recovered. Figure 6 ​​As shown, compared with the sham operation group, the model group and the negative control group, the apoptosis of neurons in the cerebral cortex of the mice was significantly increased after 24 h of cerebral ischemia reperfusion; after the intervention treatment of sh-AU020206, the apoptosis of neurons induced by MCAO / R was significantly reduced.

[0050] 9. Quantitative real-time polymerase chain reaction (qRT-PCR)

[0051] Total RNA was extracted using TRIzol reagent (Generay Biotech) according to the manufacturer's protocol. Transcription reactions were performed at 16 °C for 30 min, followed by incubation at 42 °C for 30 min, and enzyme inactivation at 85 °C for 5 min. Fast quantitative PCR was performed using SYBRHSelect Master Mix (Vazyme). These transcription reactions were performed using the following parameters: incubation at 16 °C for 30 min, incubation at 42 °C for 30 min, and incubation at 84 °C for 5 min. The qRT-PCR reaction was performed using the following parameters: incubation at 95 °C for 2 min, followed by 40 cycles of 10 s at 95 °C and 20 s at 60 °C. All results were normalized to the expression of GAPDH. Quantitative analysis was performed by using 2 -ΔΔCt Technical Methods. All primer sequences used are shown below:

[0052] lncRNA-AU020206, 5'-AGTGGTGATGAGGTGCTGTT-3' and 5'-CTGAGGTAGTCTCCAGGTGC-3'; SLC7A11, 5'-AGACGGTGGCAGTGTTTGTA-3' and 5'-TGGGTTCTTCTGGGATGACA-3'; GPX4, 5'-TGGAGCCACGCATTTGTCAT-3' and 5'-TCGTTCTTCAGGGACAGGAG-3'; YTHDC2, 5'-CCATCTTCGACTCGCTGTTC-3' and 5'-TGACTCGCTTGTTGTGGGTA-3'; TFRC, 5'-TGCCTTGTGTATGCTCCACT-3' and 5'-CAGGGAGCTGTAGGAAGGTG-3'; ACSL4, 5'-GATGACTTCGGGATCGTGGT-3' and 5'-ACAGTCTGGGACCGAAAGGT-3'; and GAPDH, 5'-AACGATTTGGTTATTG-3' and 5'-GGAAGATGTGGTATT-3'. As shown in FIG. 6, compared with the sham operation group, the model group and the negative control group, the expression of lncRNA-AU020206 was significantly reduced in the cerebral cortex of the mice after 24 h of cerebral ischemia reperfusion; after the intervention treatment of sh-AU020206, the expression of lncRNA-AU020206 was significantly increased in the cerebral cortex of the mice. Figure 7As shown, after 24h of cerebral ischemia reperfusion, compared with the sh-NC group, after sh-AU020206 intervention treatment, the mRNA expression of ferroptosis genes (TFRC, ACSL4) was significantly reduced, and the mRNA expression level of anti-ferroptosis genes (GPX4, SLC7A11) was significantly increased.

[0053] 10. Western blotting

[0054] The mice were anesthetized by intraperitoneal injection of 10% chloral hydrate solution, decapitated and killed on ice, and the cerebral hemispheres were divided into the injured side and the non-injured side. Fresh brain tissue was taken from the perifocal area of the injured side, and the brain tissue and lysis solution were fully homogenized at a mass-volume ratio of 1:10. The homogenate was placed in an ice box, shaken on a shaker for 30 min, and then centrifuged at 4°C at 12,000 rpm for 15 min. The supernatant was aspirated, 6x protein loading buffer was added at a volume ratio, denatured at 100°C for 15 min, aliquoted and stored at -80°C. The protein samples in the -80°C were taken out, dissolved and centrifuged, and then added to the lanes using a microsyringe. SDS-polyacrylamide constant pressure gel electrophoresis separation was performed, and the electric transfer system was used to transfer to the PVDF membrane. 10% skim milk powder-TBST was blocked at room temperature for 1-2h. The primary antibody was incubated on a shaker at 4°C overnight. The next day, the primary antibody was discarded, the membrane was washed with TBST for 10 min x 4 times, the horseradish peroxidase-labeled secondary antibody was added, and the shaker was incubated at room temperature for 1h. After washing with TBST, ECL luminescent substrate was added for color development. Tanon5200 full-automatic chemiluminescence imaging analysis system was used for development and analysis. The ratio of the gray value of the target protein to the gray value of the internal reference GAPDH was semi-quantitatively analyzed. Figure 8 As shown, after 24h of cerebral ischemia reperfusion, compared with the sh-NC group, after sh-AU020206 intervention treatment, the protein expression of ferroptosis genes (TFRC, ACSL4) was significantly reduced, and the protein expression level of anti-ferroptosis genes (GPX4, SLC7A11) was significantly increased.

[0055] In summary, down-regulating lncRNA-AU020206 is beneficial to avoid the occurrence of cerebral ischemia / reperfusion injury, has a protective effect on the acute stage of focal cerebral ischemia reperfusion in mice, can be widely applied in the preparation of drugs for treating ischemic stroke, has a good application prospect, and provides a direction and technical basis for finding and developing new effective drugs for treating ischemic stroke and other cerebrovascular diseases, which is beneficial to the early recovery of patients with ischemic stroke and other cerebrovascular diseases from pain and suffering.

[0056] Example 2:

[0057] The reagent for detecting the expression level of lncRNA-AU020206 in vitro is used for preparing a diagnostic kit for ischemic stroke. The reagent comprises specific primers or probes for lncRNA-AU020206. Those skilled in the art can easily design primers for amplifying the molecular marker or probes for identifying the molecular marker according to the molecular genetic marker of the present application, thereby being used for the diagnosis of ischemic stroke, for example, by PCR amplification, using 2 -ΔΔCt The relative expression amount of lncRNA-AU020206 is calculated by a technical method, and compared with that of a healthy control group.

[0058] The above description is a detailed description of the preferred and feasible experimental examples of the present application, but the examples are not used to limit the scope of the patent application of the present application. Any equivalent changes or modifications made under the technical spirit of the present application should belong to the patent scope covered by the present application.

Claims

1. Application of lncRNA-AU020206 in a stroke treatment target.

2. Application of an inhibitor for inhibiting functional expression of lncRNA-AU020206 in preparation of a drug for treating stroke.

3. Use according to claim 1 or 2, characterized in that, The nucleic acid sequence of the lncRNA-AU020206 is shown as SEQ ID NO.

1.

4. The use according to claim 2, wherein the compound is ###0002### The inhibitor is shRNA for targeting and silencing expression of lncRNA-AU020206; the nucleic acid sequence of the shRNA is shown as SEQ ID NO.

2.

5. A pharmaceutical composition, characterized by, An effective amount of the lncRNA-AU020206 inhibitor and a pharmaceutically acceptable carrier are used for treating ischemic stroke.

6. The pharmaceutical composition of claim 5, wherein, The dosage form of the pharmaceutical composition is oral preparation or injection preparation.

7. Application of a reagent for detecting expression level of lncRNA-AU020206 in vitro in preparation of a diagnostic kit for ischemic stroke.

8. Use according to claim 7, wherein the compound is ###0002### The reagent includes specific primers or probes for lncRNA-AU020206.

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