Application of miR-153-3p and interferent thereof and product

A technology for interferers and products, applied in the field of applications and products of miR-153-3p and its interferers

Pending Publication Date: 2019-11-26
QINGDAO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although more and more miRNAs have been discovered as biomarkers and therapeutic targets of human diseases, the key miRNAs and their mechanisms in heart diseases are still rarely studied, and further discoveries are needed

Method used

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  • Application of miR-153-3p and interferent thereof and product
  • Application of miR-153-3p and interferent thereof and product
  • Application of miR-153-3p and interferent thereof and product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Example 1 Detection of miRNA-153-3p expression at the cellular level during ISO-induced cardiomyocyte hypertrophy

[0091] First, we detected that during the process of ISO-induced cardiomyocyte hypertrophy, the expression level of miR-153-3p was continuously increased ( figure 1 ). In this example, the primary cardiomyocytes of rat suckling rats were cultured using the method established in this laboratory, and the hypertrophy model of cardiomyocytes was established by using the primary cardiomyocytes of rat suckling rats treated with ISO, specifically, in a cell culture incubator Rat suckling rat primary cardiomyocytes were cultured, the culture scale was 10cm culture dish, and the cell volume of each culture dish was about 1×10 6 , and the incubation time lasted 0, 8, 18, and 24 hours. RNA was extracted with different culture times, and the mRNA expression level of miRNA-153-3p was detected by real-time fluorescent quantitative PCR technology.

[0092] Specificall...

Embodiment 2

[0099] Example 2 The experiment of miRNA-153-3p antisense nucleotide effectively inhibiting cardiomyocyte hypertrophy

[0100] 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-153-3p antisense nucleotide (SEQ ID NO.2) transfection treatment (transfected by liposome lipfectin3000 according to the kit operation instructions, purchased from Invitrogen Company), after 24 hours of transfection, in In cell culture, the cells were subjected to ISO culture treatment for 24 hours to induce cell hypertrophy. At the same time, primary cardiomyocytes without miRNA-153-3p antisense nucleotide transfection were used as a negative control to detect the expression level of endogenous miRNA-153-3p, which showed that miRNA-153-3p antisense nucleotide could Effectively inhibit the endogenous expression of miRNA-153-3p ( Figure 2A ). And carry out the detection of myocardial hypertrophy ...

Embodiment 3

[0101] Example 3 miRNA-153-3p antisense nucleotides effectively inhibit the hypertrophy of cardiomyocytes in a mouse model

[0102] The antisense nucleotides (SEQ ID NO.2) of miRNA-153-3p used in this experiment are all modified miRNA-153-3p antisense nucleotides, and the modification method is to miRNA-153-3p antisense core Each base of the nucleotide was modified with 2'-methoxy group to increase the stability of miRNA-153-3p antisense nucleotide in vivo. The modification and synthesis of miRNA-153-3p antisense nucleotides were completed by Shanghai Gemma Company.

[0103] Coronary artery injection or tail vein injection of miRNA-153-3p antisense nucleotides in mice can significantly inhibit cardiomyocyte hypertrophy. Taking C57 mice as the experimental subjects, inject miRNA-153-3p antisense nucleotides and its negative control (NC) as follows

[0104] (5'-CAGUACUUUUGUGUAGUACAA-3' (SEQ IDNO: 7), and each base has been modified with 2'-methoxy, synthesized by Shanghai Gemm...

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Abstract

The invention relates to the biotechnology field and particularly provides application of miR-153-3p and interferent thereof and a product. Research finds for the first time that miR-153-3p participates in regulating cardiomyocyte hypertrophy by forming a signal path of nfatc3 / miR-153-3p / Mfn1 and affecting chondriokinesis. Therefore, the miR-153-3p can be used as an early diagnosis or auxiliary diagnosis marker for heart diseases to realize early diagnosis and early treatment of the heart diseases, and a kit for detecting miR-153-3p can realized diagnosis or auxiliary diagnosis; and besides, the inventor finds that by inhabitation of the miR-153-3p, early stage of cardiomyocyte hypertrophy can be interference, and early effective prevention or treatment of the heart diseases can be realized.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to the application and products of miR-153-3p and its interferents. Background technique [0002] Cardiac hypertrophy is a compensatory response of the myocardium to sustained increases in cardiac load and is characterized by increased cardiomyocyte volume / surface area, increased heart weight, stromal cell proliferation, and myocardial remodeling. Pathological cardiac hypertrophy is associated with ventricular remodeling and cardiac dysfunction, which is caused by many cardiovascular diseases. Cardiac hypertrophy involves not only abnormal increases in myocardial tissue and ventricular wall / septum, but also concomitant upregulation of embryonic genes, myocardial fibrosis, and cardiac insufficiency. Mitochondrial dysfunction is an important factor in the development of various cardiovascular diseases through increased production of reactive oxygen species and mitochondria...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61P9/10A61P9/00A23L33/10
CPCA61K31/7105A61P9/10A61P9/00A23L33/10A23V2002/00A23V2200/326
Inventor 王昆周露玙
Owner QINGDAO UNIV
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