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Circular RNA marker for detecting cerebral ischemia-reperfusion injury and application

A technology of cerebral ischemia-reperfusion and molecular markers, applied in the direction of DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of complex pathogenesis and multiple influencing factors of cerebral ischemia-reperfusion injury , to achieve the effect of improving clinical treatment effect, quantitative accuracy, good application value and prospect

Pending Publication Date: 2019-10-29
昆明医科大学第二附属医院
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Problems solved by technology

[0005] The present invention provides a molecular marker for detecting cerebral ischemia-reperfusion injury, and its application and use method as a detection reagent. The problem of realizing accurate diagnosis and judging the degree of disease through a single specific marker

Method used

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  • Circular RNA marker for detecting cerebral ischemia-reperfusion injury and application
  • Circular RNA marker for detecting cerebral ischemia-reperfusion injury and application
  • Circular RNA marker for detecting cerebral ischemia-reperfusion injury and application

Examples

Experimental program
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Effect test

Embodiment 1

[0023] Example 1 Total RNA extraction, circRNA sequencing and screening

[0024] Prepare rat suture embolization MCAO / R cerebral ischemia-reperfusion injury model, collect peripheral blood, select 3 groups of peripheral blood samples for inspection, and use peripheral blood samples of normal rats as a control for RNA extraction and sequencing;

[0025] The specific method is as follows: 1. According to the instructions, use TRIZOL reagent from Thermo Fisher to extract 5 μg Total RNA from each sample, then use Epicentre Ribozero rRNA Removal Kit to remove rRNA kit, and use RNase R to digest the linear RNA in it , using Illumina’s directional library preparation kit to generate a sequence library. After cluster generation, it was sequenced on a high-throughput 4000 sequencing platform to generate 150-bp double-ended reads. The above sequencing and circRNA / mRNA analysis were all performed by Novogene source company.

[0026] 2. RT-qPCR verification

[0027] Use the RevertAid Fi...

specific Embodiment 2

[0035] Specific Example 2 Identification and detection of three corresponding circular circRNA 0002086, circRNA0001606 and circRNA 0002023 in human cerebral ischemia-reperfusion blood.

[0036] Methods: In this example, 28 peripheral blood samples from patients with cerebral ischemia-reperfusion injury and 28 samples from normal subjects were collected, and the levels and specificities of circRNA 0002086, circRNA 0001606 and circRNA 0002023 were detected.

[0037] It was found that, if Figure 2-3 As shown, the expression level of circRNA 0002086 in patient samples was significantly higher than that in the normal group, the difference was statistically significant (P<0.001), and the specificity of circRNA 0002086 as cerebral ischemia-reperfusion injury was 87%.

[0038]The results showed that in the blood of patients with cerebral ischemia-reperfusion injury, the expression level of circRNA 0002086 was also significantly higher than that of healthy people. Therefore, circRNA 0...

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Abstract

The invention provides a circular RNA molecule marker circRNA 0002086 for detecting cerebral ischemia-reperfusion injury. According to the circular RNA molecule marker circRNA 0002086, a circRNA molecule marker diagnostic model is used for diagnosing the cerebral ischemia-reperfusion injury for the first time, the good diagnostic efficiency is achieved, serum level circRNA of the cerebral ischemia-reperfusion injury is detected through a real-time fluorescence quantification PCR technology, the quantification is accurate, and convenience and reliability are achieved; and the provided circRNA can be used for clinical diagnosis of the cerebral ischemia-reperfusion injury, the diagnostic specificity of the circRNA 0002086 reaches 87%. According to the circular RNA molecule marker circRNA 0002086 for detecting the cerebral ischemia-reperfusion injury, a foundation is laid for that the state of a patient with the cerebral ischemia-reperfusion injury is quickly and accurately grasped by clinical doctors, and the clinical therapeutic effect is improved, help is provided for finding of novel small molecule drug targets with potential therapeutic value, and the circular RNA molecule markercircRNA0002086 has good application value and prospects.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a circular RNA marker for detecting cerebral ischemia-reperfusion injury. Background technique [0002] Cerebral ischemia-reperfusion injury (CIR) refers to the phenomenon that the ischemic injury is further aggravated after the recanalization of the cerebral blood vessels that cause the blood flow interruption and occlusion of the brain tissue due to various reasons. It is a complex pathophysiological process involving many factors. At present, the mechanism of CIR mainly includes cell apoptosis theory, inflammatory response theory, excitatory amino acid theory and free radical theory. Inflammatory response and cell apoptosis are the main factors of nerve cell damage after CIR induction. There is no ideal brain Molecular markers for the detection of ischemia-reperfusion injury. Because the pathogenesis of cerebral ischemia-reperfusion injury is complicated and there are many influen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/166C12Q2600/178C12Q2600/136
Inventor 闵晓黎
Owner 昆明医科大学第二附属医院
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