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Application of miR-144 and inhibitor thereof

A 1. mir-144, inhibitor technology, application in the preparation of drugs for the treatment of heart disease, the field of miR-144 and its inhibitors

Inactive Publication Date: 2015-04-29
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] At present, there are no literature reports on the relationship between miR-144 and myocardial oxidative stress injury at home and abroad.

Method used

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  • Application of miR-144 and inhibitor thereof
  • Application of miR-144 and inhibitor thereof
  • Application of miR-144 and inhibitor thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1, microRNAs expression analysis in diabetic myocardial injury model

[0069] There are a variety of high-abundance microRNAs in myocardial tissue. In this experiment, the classic STZ induction method was used to successfully establish a mouse model of diabetic myocardial injury. The construction method is as follows:

[0070] Male C57BL / 6 mice, 8 weeks old, were randomly divided into two groups. Body weight was weighed after fasting for 12 hours before modeling, and fasting blood glucose was measured. One group was intraperitoneally injected with streptozotocin (STZ) solution (prepared with pH 4.5 citric acid-sodium citrate buffer), and each mouse was injected with 150 mg / kg once; the negative control group was injected with the same volume intraperitoneally Citric acid-sodium citrate buffer. One week later, the blood was collected from the tail vein, the blood glucose was measured by the blood glucose meter, and the blood glucose ≥ 18.6mol / L and positive ur...

Embodiment 2

[0076] Example 2, miR-144, a high-abundance miRNA in the heart, is mainly expressed in cardiomyocytes

[0077] Cardiomyocyte culture: Digest with 0.2% trypsin and 0.1% collagenase, purify cardiomyocytes by differential adhesion and attachment method, culture in IMDM medium, observe under an inverted microscope, and evaluate the quality of cardiomyocytes by trypan blue staining , Myocytes were analyzed by Actin immunohistochemistry, Myoglobin fluorescence immunohistochemistry and electron microscopy.

[0078] Myocardial fibroblast culture: primary culture of mouse cardiac fibroblasts by enzyme digestion + differential attachment method. At the bottom of the bottle, fibroblasts were obtained by repeated differential attachment for 1 hour, and the cell morphology was observed and immunocytochemical staining was used to verify the purity of the cells (cardiac fibroblasts showed a strong positive reaction for vimentin and a negative reaction for actin).

[0079] The present invent...

Embodiment 3

[0081] Example 3, the expression of miR-144 gradually increases in the pathological process of diabetic myocardial injury

[0082] The method is the same as example 1 and 2

[0083] In the previous work, we detected the expression of microRNAs in the left ventricle tissue of the diabetic myocardial injury mouse model by quantitative PCR, and found that the abnormal expression of miR-144 was more obvious and there was a relatively specific myocardial expression; and the plasma miR-144 level was also higher than that of the control group. Significantly increased. We will further explore the abnormal expression of miR-144 in the whole process of myocardial oxidative stress, and explore its possible pathological significance. The results showed that: miR-144 did not change at 1 week, the expression of miR-144 began to increase to 1.5 times that of the sham group at 4 weeks, and the expression level was about 3 times that of the sham group at 12 weeks, suggesting that miR-144 may ...

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Abstract

The invention relates to the technical field of biological medicines. That miR-144 can specifically inhibit the gene expression of Nrf2 is disclosed for the first time, the expression of miR-144 is closely related to heart disease, and the high expression of miR-144 can promote myocardial oxidative stress to cause myocardial damage related diseases. The invention provides miR-144 and application of an inhibitor of miR-144 in preparation of medicines for preventing or treating myocardial oxidative stress and damage diseases, especially diabetes caused myocardial oxidative stress and damage diseases; a new target spot is provided for the diagnosis and prevention and treatment of heart diseases.

Description

technical field [0001] The invention relates to the research field of biotechnology microRNA, more specifically, relates to the application of miR-144 and its inhibitors in the preparation of drugs for treating heart diseases. Background technique [0002] MicroRNA is a kind of small molecule RNA with post-transcriptional regulatory activity discovered in Drosophila, nematode, mouse and human in recent years. The discovery of a large number of endogenous small RNAs benefits from two technologies, one is the construction and sequencing of microRNA cDNA libraries; the other is the capture of biotin-labeled oligonucleotide probes, which are A technique for performing PCR amplification. Through library construction and sequencing, people have mastered a large amount of microRNA sequence information, and through bioinformatics comparison analysis of these sequences, it has been found that most microRNAs are highly conserved among species and highly homologous in evolution. [0...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K45/00A61K31/7088C12Q1/68A61P9/04A61P3/10
Inventor 于曼丽刘宇秦永文赵仙先
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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