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Application of piRNA-5938 and antisense nucleic acid thereof in diagnosis, prevention, treatment and prognosis evaluation of ischemic heart disease

A pirna-5938, antisense nucleic acid technology, applied in cardiovascular system diseases, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as lack of effective cure for ischemic heart disease

Active Publication Date: 2017-11-24
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective means of curing ischemic heart disease clinically

Method used

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  • Application of piRNA-5938 and antisense nucleic acid thereof in diagnosis, prevention, treatment and prognosis evaluation of ischemic heart disease
  • Application of piRNA-5938 and antisense nucleic acid thereof in diagnosis, prevention, treatment and prognosis evaluation of ischemic heart disease
  • Application of piRNA-5938 and antisense nucleic acid thereof in diagnosis, prevention, treatment and prognosis evaluation of ischemic heart disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] The expression of piRNA-5938 is increased in the hearts of C57 mice treated with ischemia-reperfusion after ligation of the left anterior descending coronary artery

[0073] 1. Establishment of ischemia-reperfusion model of ligated left anterior descending coronary artery

[0074] 1) C57 mice were weighed and anesthetized with 2% chloral hydrate at a dose of 0.2mL / 10g;

[0075] 2) After the mouse is fully anesthetized, the mouse is fixed on the operating table, and the electrocardiogram is connected at the same time to detect the heartbeat;

[0076] 3) the mouse chest and neck hair were shaved, and iodophor was applied for disinfection;

[0077] 4) Tracheal intubation, cut the skin in the middle of the mouse neck, then separate the fat layer and muscle with tweezers, expose the trachea and thread it under the trachea; use surgical scissors to cut a small opening above the trachea, and place the ventilator The cannula was inserted into the trachea, fixed with surgical ...

Embodiment 2

[0113] piRNA-5938 expression is increased in primary cardiomyocytes treated with hydrogen peroxide

[0114] 1. Isolation of primary cardiomyocytes

[0115] 1) Equipment for autoclaving experiments, including scissors, tweezers, blades, glass plates, 260-mesh filter, 50mL Erlenmeyer flask, small rotor, tip, pipette; prepare all reagents for the experiment; prepare ice box, 37°C air bath shaker;

[0116] 2) Treat the petri dish on which cardiomyocytes will be cultured with Poly-L-lysin at 37°C for 1 hour, then wash 2-3 times with sterile three-distilled water, and dry in the air; the function of Poly-L-lysin is to make the petri dish Facilitate the adherence of cardiomyocytes;

[0117] 3) Prepare an ice box in the ultra-clean bench, put 4 petri dishes filled with 1×ADS buffer on the ice box to pre-cool;

[0118] 4) To kill rats, put 1-2 day old SD rat suckling rats, regardless of sex, into 75% alcohol for cleaning and disinfection twice, and finally drown them;

[0119] 5) G...

Embodiment 3

[0139] piRNA-5938 expression is increased in hypoxia-treated primary cardiomyocytes

[0140] 1. The primary cardiomyocytes are separated, and the operation method is the same as the step in Example 2;

[0141] 2. Hypoxia treatment of primary cardiomyocytes

[0142] Rat suckling mouse primary cardiomyocytes were cultured in a hypoxic incubator (oxygen concentration lower than 1%), and the culture scale was 10 cm culture dish, and the cell volume of each culture dish was about 1×10 6 Each, the culture time lasts 0, 10min, 30min, 60min;

[0143] 3. Total RNA was extracted, and the method of operation was the same as the step in Example 1;

[0144] 4. The cDNA preparation of piRNA-5938, the operation method is the same as the step in Example 1;

[0145] 5. Real-time fluorescent quantitative PCR detects the expression level of piRNA-5938, and the operation method is the same as the step in Example 1;

[0146] The result is as image 3 as shown, image 3 In order to isolate th...

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Abstract

The invention provides application of piRNA and antisense nucleic acid thereof in diagnosis, prevention, treatment and prognosis evaluation of an ischemic heart disease. The piRNA provided by the invention is a nucleic acid containing a sequence shown in SEQ ID No.2 or a bioactive functional fragment or variant thereof; the piRNA can be taken as a new biomarker and applied to diagnosis and / or prognosis evaluation of the ischemic heart disease; besides, the antisense nucleic acid of the piRNA can be taken as a novel medicine and used for preventing and / or treating the ischemic heart disease.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and relates to a new application of an endogenous non-coding small molecule RNA and its antisense nucleic acid, in particular to a piRNA (piwi-interacting RNA) and its antisense nucleic acid in diagnosis and / or Use in the prevention and / or treatment and / or prognosis assessment of ischemic heart disease. Background technique [0002] Cardiovascular disease is a major disease that threatens human life and health, and its morbidity and mortality rank first among all diseases. At present, the number of patients with cardiovascular diseases in China is about 290 million, and the prevalence and mortality are still rising. The annual medical expenses for cardiovascular diseases are as high as 130 billion yuan, which imposes a huge economic and social burden on the country and the people. Therefore, it is of great significance to strengthen the research of heart disease, and its research results w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11A61K31/7105A61P9/10A61P9/04
CPCA61K31/7105C12Q1/6883C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 李培峰孙腾王建勋范长江于万鹏常文光
Owner QINGDAO UNIV
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