MicroRNA-328 and application of antisense nucleotide thereof for diagnosing, preventing and curing heart diseases

A heart disease, nucleotide sequence technology, applied in the direction of cardiovascular system diseases, gene therapy, microbial determination/testing, etc.

Inactive Publication Date: 2010-02-10
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although more and more microRNAs have been discovered as biomarkers, determinants and therapeutic targets of human diseases, key microRNA isoforms in atrial fibrillation, ischemia-reperfusion and myocardial fibrosis heart diseases are still not identified, and their function remains a challenge for researchers in the field

Method used

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  • MicroRNA-328 and application of antisense nucleotide thereof for diagnosing, preventing and curing heart diseases
  • MicroRNA-328 and application of antisense nucleotide thereof for diagnosing, preventing and curing heart diseases
  • MicroRNA-328 and application of antisense nucleotide thereof for diagnosing, preventing and curing heart diseases

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0021] Screening of miRNA expression in experimental example 1 atrial fibrillation (AF)

[0022] In this experiment, the atrial rapid pacing method was used to establish the AF model, and the pacing electrode was sutured and implanted in the right atrial appendage for pacing (400 beats / min), and Holter monitoring was performed after the operation. 27 dogs completed the entire experiment. 1 dog terminated the experiment due to pacemaker failure; 2 dogs died of pouch hematoma at 2 weeks after operation; 2 dogs died of postoperative infection; 1 dog died suddenly at 5 weeks after operation. , There are more fresh thrombi in the right atrial appendage, which is considered to be related to thromboembolism. None of the dogs induced AF by programmed stimulation before pacing.

[0023] A pacemaker was implanted in the canine atrium, and no AF occurred in all canine preprogrammed stimulation checks before pacing. After 8 weeks of rapid atrial pacing, 12 out of 17 dogs in the A-TP gr...

experiment example 2

[0027] Construction of experimental example 2 adenovirus vector

[0028] Use liposome 2000 to construct the pDC316-EGFP-U6 shuttle plasmid (please give the specific construction method of "pDC316-EGFP-U6 shuttle plasmid"; if the specific construction method of the shuttle plasmid has been disclosed in the literature, please show The specific source of this document is sufficient) and pBHGlox backbone plasmid (please give the commercial source of "pBHGlox backbone plasmid" or the specific construction method) to co-transfect HEK-293 cells at a ratio of 7-8:1. Cytopathy, and the expression of GFP. Replace the serum-free MEM, collect the cells, freeze and thaw repeatedly 3 times to lyse the cells, centrifuge at 10,000g to collect the supernatant as the virus liquid, and use it for the infection experiment, and observe the expression of GFP after 4 days of infection ( image 3 ).

[0029] Adult healthy mongrel dogs weighing 20-30kg were selected and divided into three groups: (1...

experiment example 3

[0031] Experimental example 3 Construction of miR-328 cardiac overexpression TG mouse model

[0032] The expression of miR-328 in each organ of TG mice was detected by real time PCR, and the results showed ( Figure 6 A): Compared with non-transgenic mice (WT), the expression of miR-328 in the left atrium of TG mice was at least 17 times higher (*p Figure 6 B): All 14 positive TG mice showed spontaneous AF, that is, the P wave disappeared and the R-R interval was unequal. The above changes were not detected in WT mice.

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Abstract

The invention discloses a microRNA-328 and the application of antisense nucleotide thereof for diagnosing and controlling heart diseases; in the invention, a plurality of experiments prove that the microRNA-328 is changed and has pathopoiesia function in heavy heart diseases (atrial fibrillation, ischemia-reperfusion or myocardial fibrosis), and the antisense nucleotide AMO-328 of the microRNA-328has prevention and cure functions to the heart diseases, the microRNA-328 is used as a novel drug effect target spot, so as to finally determine and detect that the peripheral blood microRNA-328 canbe used as a novel biological marker which is used for diagnosing and preventing the heart diseases; in addition, the peripheral blood microRNA-328 expression level of a patient can be detected through fluorescent quantitative PCR and the nanometer microballoon technology enhanced by silver staining, so as to be used for diagnosing and preventing the heart diseases such as atrial fibrillation, long QT syndrome, ischemia-reperfusion or myocardial fibrosis and the like.

Description

technical field [0001] The present invention relates to a new medical application of endogenous non-coding small RNAs, in particular to the application of microRNA-328 and its antisense nucleotides in the diagnosis and prevention of heart disease, belonging to the field of diagnosis, prevention and treatment of heart disease . Background technique [0002] Heart disease is a highly prevalent disease with high morbidity, mortality, and resulting socioeconomic burden. Regarding the prevention, diagnosis and treatment of heart disease, there are still huge gaps. Atrial fibrillation, ischemia-reperfusion arrhythmia, and long QT syndrome are the main manifestations of clinical heart disease. The current treatment effect is not good, and the mortality rate is high. [0003] As a major pathological cause, myocardial fibrosis is involved in many heart diseases such as heart failure and arrhythmia. The pathogenesis of heart disease is complex, each pathogenesis can lead to one or ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K48/00A61P9/00A61P9/06A61P9/10
Inventor 杨宝峰王志国吕延杰单宏丽王宁张莹潘振伟初文峰
Owner HARBIN MEDICAL UNIVERSITY
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