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Composition for inhibition of expression of miR-27b, medicines containing the same, and use for the same

A technology of mir-27b and compound, which is applied in the directions of medical preparations, drug combinations, and pharmaceutical formulations containing active ingredients, can solve the problems of unreported specific heart function and elevated expression level, and achieve the relief of cardiac systolic function and ease of use. and fibrosis, relieve myocardial hypertrophy

Inactive Publication Date: 2011-10-05
INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Previous studies have shown that the expression level of miR-27b is significantly increased in mouse models of cardiac hypertrophy and human specimens of aortic sclerosis, suggesting that it may play an important role in the occurrence of cardiac diseases, but regarding its role in the heart The specific function has not yet been reported

Method used

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  • Composition for inhibition of expression of miR-27b, medicines containing the same, and use for the same
  • Composition for inhibition of expression of miR-27b, medicines containing the same, and use for the same
  • Composition for inhibition of expression of miR-27b, medicines containing the same, and use for the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Packaging and cell infection experiments of adenovirus overexpressing miR-27b

[0042] The gene sequence containing the precursor of miR-27b was connected into the pAd-Track vector, linearized with Pme I, and recombined with the pAd-easy vector in BJ5183 bacteria to obtain pAd-miR-27b. The obtained vector was linearized and recovered by Pac I, and then transfected into 293A cells to obtain an adenovirus (pAd-miR-27b) overexpressing miR-27b. After virus amplification, infect cardiomyocytes at 100moi for 48h, (at the same time, Ad-GFP was used as a negative control, and Ad-GFP was infected and treated with phenylephrine (PE) as a positive control), through immunofluorescence staining and measurement software Measure and analyze the effect of miR-27b overexpression on the size of cardiomyocytes, extract cellular RNA, and detect changes in hypertrophy marker genes after reverse transcription. see results figure 1 , showed that miR-27b promotes cardiac hypertroph...

Embodiment 2

[0043] Example 2 Packaging and identification of knockdown miR-27b adenovirus

[0044] The reverse complementary sequence to the mature sequence of miR-27b was ligated into the pAd-Track vector, linearized with Pme I and recombined with the pAd-easy vector in BJ5183 bacteria to obtain pAd-anti-miR-27b. The obtained vector was linearized and recovered by Pac I, and then transfected into 293A cells to obtain adenovirus Ad-anti-miR-27b knocking down miR-27b. The obtained adenovirus Ad-anti-miR-27b was amplified and then infected with isolated primary cardiomyocytes, RNA was extracted after 48 hours, and miRNA northern blot detection was performed, see image 3 , found that this adenovirus could successfully knock down the expression of miR-27b.

Embodiment 3

[0045] Example 3 Establishment of TAC surgery model and detection of miR-27b expression level

[0046] (1) Establishment of the surgical model

[0047] 1. Anesthetize the animal with 0.125% tribromoethanol, and perform tracheal intubation on the mouse;

[0048] 2. Move the mouse to the dissecting table for surgery, and fix the limbs and tail of the mouse;

[0049] 3. Under the guidance of the microscope, cut the thorax of the mouse along the sternum, and use a chest opener to expand and fix a small incision, which is convenient for the subsequent ligation operation;

[0050] 4. Isolate the ascending aorta of the heart, and use a No. 26 standard marker to ligate around the ascending aorta to cause an increase in pressure load; close the chest cavity, and place the mouse on an incubator until it wakes up;

[0051] 5. The mice in the control group were subjected to the same operation, but the ascending aorta was not ligated.

[0052] (2) Detection of miR-27b expression level ...

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Abstract

The invention relates to a composition for specifically inhibiting the expression of miR-27b and medicines containing the composition. The composition has the nucleotide sequence 5' -GCAGAACUUAGCCACUGUGAA-3' of functional molecules produced in vivo. The inhibition of the expression of miR-27b in vivo can alleviate cardiac hypertrophy and heart damage caused by stress load. The invention determines the functions of miR-27b in cardiac hypertrophy through myocardial cell experiments. A mouse model of cardiac hypertrophy caused by TAC surgically created stress load is treated by packaging adenovirus for the specific inhibition of the expression of miR-27b and synthesizing an inhibitor against miR-27b, which brings out that cardiac hypertrophy and fibrosis caused by TAC operation can be alleviated effectively by respectively utilizing two inhibitors to treat the TAC postoperative mouse, with obvious thinning of ventricular wall thickness and obvious remission of systolic function of heart.

Description

technical field [0001] The invention relates to the application of miR-27b as a target molecule for heart disease treatment, in particular to a drug for treating heart disease with miR-27b as a target. Background technique [0002] In recent years, the incidence of heart disease has gradually increased, and most patients with heart disease will eventually die due to heart failure. According to the statistics of the World Health Organization, heart disease has become one of the main causes of death. The occurrence and development of heart failure is a complex process involving many factors, among which myocardial hypertrophy and myocardial fibrosis are two important factors. Myocardial hypertrophy is usually a hypertrophic reaction of the myocardium under many pathological stimuli, such as myocardial ischemia, hypertension, and heart valve defects. . Although the early compensatory period of myocardial hypertrophy is considered to be an adaptive response of the heart, which...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/713A61K48/00A61P9/00
Inventor 杨晓王剑侯宁孙强张彦
Owner INST OF BIOENG ACAD OF MILITARY MEDICAL SCI OF THE CHINESE
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