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Novel application of miRNA-484 and pharmaceutical composition containing miRNA-484 and its application

A technology of mirna-484, 1.mirna-484, applied in the field of biopharmaceuticals, can solve the problems that the pathogenesis of heart disease is not completely clear, the prevention, diagnosis and treatment of heart disease cannot achieve satisfactory results, etc.

Inactive Publication Date: 2011-12-07
INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • 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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  • Novel application of miRNA-484 and pharmaceutical composition containing miRNA-484 and its application
  • Novel application of miRNA-484 and pharmaceutical composition containing miRNA-484 and its application
  • Novel application of miRNA-484 and pharmaceutical composition containing miRNA-484 and its application

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0028] Experimental example 1 Myocardial ischemia and myocardial cell apoptosis induced by hypoxia miRNA-484 expression detection

[0029] For the experimental model of cardiomyocyte apoptosis, the primary rat neonatal cardiomyocytes were cultured using the method established in our laboratory (the rat neonatal rat was purchased from Beijing Medical University, the rat strain is Wistar rat, the rat neonatal primary rat The preparation of cardiomyocytes can be found in the following documents: W.-Q. Tan, et al, Foxo3a Inhibits Cardiomyocyte Hypertrophy through Transactivating Catalase J Biol Chem. 2008 October 31; 283(44): 29730-29739), hypoxic incubator (low oxygen concentration) In 1%) culture at different times, extract RNA, and detect the expression level of miRNA-484 by real-time fluorescent quantitative PCR technology. Such as figure 1 As shown in A, the expression level of miRNA-484 was significantly down-regulated within 10min~180min of hypoxia treatment.

[0030] In this...

experiment example 2

[0031] Experimental example 2 miRNA-484 inhibits cardiomyocyte apoptosis

[0032] The miRNA-484 adenovirus was constructed using the pSilencer Adeno 1.0-CMV system from Ambion. Construct the corresponding vector and adenovirus according to the company's instructions. The specific method is as follows: extract the genome of rat heart, use it as a template, PCR amplify miRNA-484 and its upstream and downstream ends each with nearly 200 bp sequence (PCR system 50μL, The PCR conditions are as follows: 95°C for 3min for a cycle; 95°C for 30s, 56°C for 30s, 72°C for 1min in 28 cycles; finally 72°C for extension for 5min), the amplified fragment is double digested with spe I and xho I, and then The adenoviral vector pSilencer Adeno CMV1.0 shuttle vector (pSilencer Adeno CMV1.0 shuttle vector) (purchased from Ambion) was double-enzyme digested and then ligated with the digested fragment, that is, the fragment was cloned into the adenovirus vector. Pac I digestion and linearization of t...

experiment example 3

[0034] Experimental example 3 miRNA-484 overexpression inhibits myocardial ischemia injury experimental verification

[0035] Mice injected miRNA-484 mimics via coronary artery can significantly inhibit myocardial ischemia-reperfusion injury. C57 / BL6 mice were used as experimental subjects, and miRNA-484 mimics (miRNA-484mimic) and miRNA-484 mimic negative controls (miRNA-484mimic NC) were injected as follows, and sham-operated mice were used as control groups , C57 / BL6 mice are wild type. The mice were weighed and anesthetized, fixed in the supine position, sheared and disinfected the operation area with iodine. Cut the trachea in the middle of the neck and intubate, connect the animal ventilator for positive pressure ventilation, cut the skin about 3cm longitudinally at the left edge of the sternum and the heart beat, and bluntly separate the subcutaneous tissue and muscle layer by layer, open the chest, and be careful Lift the pericardium and cut it open to fully expose the...

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Abstract

The invention provides a new application of miRNA-484, a pharmaceutical composition containing miRNA-484 and the application thereof. The use of the miRNA-484 in the preparation of medicines for preventing or treating heart disease, the pharmaceutical composition, the pharmaceutical composition is composed of an effective dose of miRNA-484 pharmaceutically acceptable carrier, virus or auxiliary material. miRNA-484 has a protective effect on the heart by inhibiting cardiomyocyte apoptosis, and miRNA-484 has potential preventive and therapeutic value for many heart diseases.

Description

Technical field [0001] The present invention relates to a new use of endogenous non-coding small RNAs, in particular to the use of mRNA-484 in the prevention or treatment of heart diseases, belonging to the technical field of biopharmaceuticals. Background technique [0002] Cardiovascular disease is the main threat to human health and life, and the number one killer of human health. Seventeen million people die of cardiovascular disease worldwide each year, and its mortality rate is close to the sum of all cancer mortality. In my country, with the increasing living standards of the people and changes in dietary structure, the death rate of cardiovascular diseases is showing a clear upward trend, and it has surpassed cancer as the leading cause of death. According to the latest statistics published by the Ministry of Health at the end of 2004, the prevalence rate of hypertension among residents aged 18 and over in my country is 18.8%. It is estimated that the number of patients i...

Claims

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Application Information

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