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MiRNA-361 (micro-ribonucleic acid-361) and application of antisense nucleotide thereof

A miRNA-361, antisense nucleotide technology, applied in the determination/examination of microorganisms, gene therapy, cardiovascular system diseases, etc., can solve the problem that the prevention, diagnosis and treatment of heart disease cannot achieve satisfactory results, heart disease The pathogenesis of the disease is not fully understood, etc.

Active Publication Date: 2013-02-13
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the pathogenesis of heart disease is not completely clear, and the prevention, diagnosis and treatment of heart disease cannot achieve satisfactory results. It is urgent to develop new technologies and methods for the diagnosis, prevention and treatment of heart disease.

Method used

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  • MiRNA-361 (micro-ribonucleic acid-361) and application of antisense nucleotide thereof
  • MiRNA-361 (micro-ribonucleic acid-361) and application of antisense nucleotide thereof
  • MiRNA-361 (micro-ribonucleic acid-361) and application of antisense nucleotide thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Detection of miRNA-361 expression in cardiomyocyte hypoxia and myocardial ischemia-induced apoptosis

[0034]In this example, the method established in this laboratory was used to culture primary cardiomyocytes of rat neonatal rats for the construction of an experimental model of cardiomyocyte apoptosis. For the specific steps of preparing primary cardiomyocytes of rat neonatal rats, please refer to the literature : W.-Q. Tan, Kun Wang, et al., Foxo3a Inhibits Cardiomyocyte Hypertrophy through Transactivating Catalase. J Biol Chem. 2008 October 31;283(44):29730-29739.

[0035] The myocardial hypoxia model was established by using the primary cardiomyocytes of rat suckling rats. Specifically, the primary cardiomyocytes of rat suckling rats were cultured in a hypoxic incubator (oxygen concentration lower than 1%), and the culture scale was a 10cm petri dish. , the cell volume per dish is about 1 × 10 6 , and the incubation time lasted 0, 1, 2, 3, 4 hours. RNA ...

Embodiment 2

[0042] Example 2 The experiment of miRNA-361 antisense nucleotides inhibiting cardiomyocyte apoptosis

[0043] In this experiment, the primary cultured cardiomyocytes in Example 1 were used as a model. For cardiomyocytes (the cell volume is about 1×10 6 ) for miRNA-361 antisense nucleotide transfection treatment (transfected by liposome lipfectin2000 according to the kit operation instructions, purchased from Invitrogen Company), 24 hours after transfection, in a hypoxic incubator (oxygen concentration lower than 1%) for 3 hours to induce cell apoptosis. At the same time, primary cardiomyocytes without miRNA-361 antisense nucleotide transfection were used as a negative control, and apoptosis of cardiomyocytes was detected by trypan blue staining.

[0044] Experimental results such as figure 2 As shown, it was shown that inhibition of endogenous miRNA-361 could significantly inhibit hypoxia-induced cardiomyocyte apoptosis.

Embodiment 3

[0045] Example 3 miRNA-361 antisense nucleotide inhibits myocardial ischemia injury experiment

[0046] Coronary injection of miRNA-361 antisense nucleotides in mice can significantly inhibit myocardial ischemia-reperfusion injury. Taking C57 mice as the experimental subjects, inject miRNA-361 antisense nucleotides and its negative control (5'-CAGUACUUUUGUGUAGUACAA-3' (SEQ ID NO: 7) according to the following method, and each base was 2' -Methoxy modification, synthesized by Shanghai Jima Company,) (NC): Mice were weighed and anesthetized, fixed in supine position, sheared and disinfected with iodine in the operation area. The trachea was cut midline in the neck and intubated, and the animal ventilator was used for positive pressure ventilation. The skin was cut longitudinally about 3 cm at the left edge of the sternum and the beating heart, and the subcutaneous tissue and muscles were bluntly separated layer by layer. Open the chest and be careful. Lift the pericardium and c...

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Abstract

The invention discloses miRNA-361 (micro-ribonucleic acid-361) and application of antisense nucleotide thereof, and in particular discloses application of miRNA-361 antisense nucleotide sequence in preparing a medicament for preventing and treating heart diseases and application of miRNA-361 ribonucleic acid in preparing a medicament or kit for diagnosing or prognosing heart diseases. Massive experiments prove variation of the miRNA-361 antisense nucleotide in myocardial ischemial injury and myocardium cell apoptosis and the myocardial protection effect of the miRNA-361 antisense nucleotide. The miRNA-361 antisense nucleotide has the functions of inhibiting myocardial ischemial injury and myocardium cell apoptosis, can be used as a novel drug action target, and has important meaning to prevention and treatment of myocardial fibrosis, coronary heart disease, congestive heart failure and other heart diseases caused by myocardial ischemia clinically.

Description

technical field [0001] The present invention relates to a new drug use of endogenous non-coding small RNAs, in particular to the use of miRNA-361 and its antisense nucleotides, especially to the preparation of miRNA-361 and its antisense nucleotides The invention relates to a medicine for diagnosing and preventing and treating heart diseases, belonging to the field of diagnosis, prevention and treatment of heart diseases. Background technique [0002] Cardiovascular disease is a major threat to human health and life, and is the number one killer of human health. Seventeen million people die from cardiovascular disease every year worldwide, and its mortality rate is close to the sum of all cancer mortality rates. In my country, with the improvement of people's living standards and the change of diet structure, the mortality rate of cardiovascular diseases is on the rise, surpassing cancer as the number one cause of death. According to the latest statistical results released ...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12Q1/68A61P9/00A61P9/10
Inventor 李培峰王昆
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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