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Application of microRNA-320(miR-320a) and its antisense nucleotide in diagnosis, prevention and treatment of cardiovascular diseases

A cardiovascular and cerebrovascular disease and nucleic acid technology, which is applied in the field of use of microRNA-320 (miR-320a) and its antisense nucleotides in the diagnosis and prevention of cardiovascular diseases

Active Publication Date: 2015-03-04
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether there is a class of miRNAs that play an important regulatory role in the key links or common pathways of the occurrence and development of atherosclerosis, and whether it can provide new strategies for the treatment of atherosclerosis by interfering with such miRNAs is still a research field in this field. people challenge
Some researchers have found that miR-320 is involved in the pathophysiological process of acute myocardial infarction (Circulation.2009; 119(17): 2357-66), but there is no specific subtype and miR-320 on inflammation, lipid metabolism and atherosclerosis In-depth study of the influence of sclerosis on the pathophysiological process

Method used

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  • Application of microRNA-320(miR-320a) and its antisense nucleotide in diagnosis, prevention and treatment of cardiovascular diseases
  • Application of microRNA-320(miR-320a) and its antisense nucleotide in diagnosis, prevention and treatment of cardiovascular diseases
  • Application of microRNA-320(miR-320a) and its antisense nucleotide in diagnosis, prevention and treatment of cardiovascular diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1. Screening of peripheral blood miRNA expression in patients with atherosclerosis

[0019] 1. Collect peripheral blood samples from 10 cases of atherosclerosis, 10 cases of acute myocardial infarction and 10 cases of normal controls (taken from patients hospitalized in the Department of Cardiovascular Medicine from 2005 to 2010, each case was confirmed by two or more clinicians Classification). After sampling, centrifuge at 3,500 rpm for 6 minutes at room temperature, take the upper layer of plasma, and store it in a -80°C refrigerator. Add 1ml TRIZOL LS (Invitrogen Company) to every 0.25ml of peripheral blood plasma, and homogenize with an electric homogenizer. After standing at room temperature for 5 minutes, add 0.2ml of chloroform, mix well, leave at room temperature for 5 minutes, and centrifuge at 12,000g at 4°C for 15 minutes. Transfer the upper aqueous phase to a new EP tube, add 0.5ml of isopropanol, mix well, place at room temperature for 10min, and...

Embodiment 2

[0047] Example 2. Detection of aortic miRNA-320a expression in atherosclerotic mice

[0048] 1. In this experiment, 4-week-old ApoE - / - Mice and control mice (C57BL6) were fed with a high-fat diet for 8 weeks, and were given tail vein injections of pSilencer-miR-320a plasmid and pSilencer-anti-miR-320a plasmid at a dose of 5 mg / kg body weight. Specifically divided into 6 groups: wild-type mice control (high-fat feeding C57BL6 mice), wild-type mice miR-320a treatment intervention (high-fat feeding C57BL6+pSilencer-miR-320a), wild-type mice anti-miR- 320a treatment intervention (high fat feeding C57BL6+pSilencer-anti-miR-320a), ApoE - / - Mice (high-fat feeding ApoE - / - mice), ApoE - / - Mice miR-320a treatment intervention (high-fat feeding ApoE - / - +pSilencer-miR-320a) and ApoE - / - Mice anti-miR-320a treatment intervention (high-fat feeding ApoE - / - +pSilencer-anti-miR-320a).

[0049] 2. At the end of the experiment (week), the mouse aortic RNA was routinely extracted:

[...

Embodiment 3

[0082] Example 3. The role of miR-320a and anti-miR-320a in blood lipid regulation in atherosclerotic mice

[0083] In this experiment, 4-week-old ApoE - / - Mice and control mice (C57BL6) were fed with a high-fat diet for 8 weeks, and were given tail vein injections of pSilencer-miR-320a plasmid and pSilencer-anti-miR-320a plasmid at a dose of 5 mg / kg body weight. Specifically divided into 6 groups: wild-type mice control (high-fat feeding C57BL6 mice), wild-type mice miR-320a treatment intervention (high-fat feeding C57BL6+pSilencer-miR-320a), wild-type mice anti-miR- 320a treatment intervention (high fat feeding C57BL6+pSilencer-anti-miR-320a), ApoE - / - Mice (high-fat feeding ApoE - / - mice), ApoE - / - Mice miR-320a treatment intervention (high-fat feeding ApoE - / -+pSilencer-miR-320a) and ApoE - / - Mice anti-miR-320a treatment intervention (high-fat feeding ApoE - / - +pSilencer-anti-miR-320a).

[0084] At the end of the experiment, the whole blood of the mice was collected...

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Abstract

The invention discloses application of miR-320a and its antisense nucleotide respectively in risk assessment / diagnosis of atherosclerotic cardiovascular and cerebrovascular diseases and prevention and treatment of atherosclerotic cardiovascular and cerebrovascular diseases. The inventor conducts a lot of experimental study to prove that: the miR-320a level changes (rises) in blood plasma (or serum) of high-risk groups of atherosclerotic cardiovascular and cerebrovascular diseases (atherosclerosis, coronary heart disease, myocardial infarction and stroke) and coronary heart disease, and the miR-320a level rise can induce atherosclerosis, inflammatory state and blood-lipid metabolic disorder obviously; and miR-320a's antisense nucleotide anti-miR-320a can alleviate, prevent and treat the atherosclerotic cardiovascular and cerebrovascular diseases and relevant disorder states. According to the invention, finally conclusions are obtained as the following: (1) miR-320a can be taken as a novel target for drugs, the nucleotide anti-miR-320a can be used for preventing and treating atherosclerotic cardiovascular and cerebrovascular diseases (such as atherosclerosis, coronary heart disease, myocardial infarction and stroke and the like); and (2) the peripheral blood miR-320a level can be adopted as a new biomarker, and its expression level can be detected for predicting and assessing the risk of atherosclerotic cardiovascular diseases or assisting in diagnosing already existing atherosclerotic cardiovascular diseases. Also, the fluorescent quantitative PCR (polymerase chain) technology can be employed to detect the expression level of miR-320a in patient peripheral blood so as to assist diagnosis of the atherosclerotic cardiovascular diseases.

Description

1. Technical field [0001] The present invention relates to a new medical application of endogenous small non-coding RNA, more specifically to microRNA-320 (hsa-miR-320a, SEQ ID No: 1, 5'-aaaagcuggguugagagggcga-3') and its reverse Sense nucleotide sequence hsa-anti-miR-320a (SEQ ID No: 2, 5'-tcgccctctcaacccagctttt-3') in risk assessment of atherosclerotic cardiovascular and cerebrovascular diseases and atherosclerotic cardiovascular and cerebrovascular diseases The use in the prevention and treatment of cardiovascular diseases belongs to the field of diagnosis, prevention and treatment of cardiovascular diseases. 2. Background technology [0002] Atherosclerosis is one of the pathophysiological basis of many clinical cardiovascular events, and has become the leading cause of mortality and disability. Existing studies have shown that atherosclerosis is caused by multiple factors, an inflammatory immune process involving a variety of cells, under the action of factors such as ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68A61K48/00A61P9/10A61P9/00
Inventor 汪道文陈琛杨盛兰王峰龙光文
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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