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RNA(Ribonucleic Acid)and application thereof in diseases of cardiovascular system

A technology in the heart and sequence listing, applied in cardiovascular system diseases, DNA/RNA fragments, applications, etc., can solve the problems of HIP-55 research reports and less understanding of HIP-55 functions

Inactive Publication Date: 2015-05-06
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the function of HIP-55 is poorly understood, and there is no research report on the role of HIP-55 in the cardiovascular system.

Method used

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  • RNA(Ribonucleic Acid)and application thereof in diseases of cardiovascular system
  • RNA(Ribonucleic Acid)and application thereof in diseases of cardiovascular system
  • RNA(Ribonucleic Acid)and application thereof in diseases of cardiovascular system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1. Obtaining of HIP-55shRNA, recombinant adenovirus overexpressing HIP-55 and recombinant adenovirus expressing RNA interference HIP-55

[0043] 1. Obtaining HIP-55shRNA

[0044] The following HIP-55 shRNA and control shRNA were designed and synthesized for the mouse HIP-55 protein coding gene as shown in Table 1:

[0045] HIP-55shRNA, its nucleotide sequence is sequence 3 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 4 in the sequence listing;

[0046] Design Control shRNA: its nucleotide sequence is sequence 5 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 6 in the sequence listing;

[0047] Table 1 is shRNA and its control sequence

[0048]

[0049] 2. Construction and preparation of recombinant adenovirus overexpressing HIP-55

[0050] 1. Construction of recombinant adenoviral vector for overexpressing HIP-55

[0051] Amplification of attB1-HIP-55-Flag-IRES / DesRed-attB2 by Overla...

Embodiment 2

[0178] Example 2, Functional Research of HIP-55

[0179] 1. The effect of HIP-55 on the proliferation of cardiac fibroblasts

[0180] 1) Brdu ELISA detection

[0181] The isolated cardiac fibroblasts were subcultured in a 96-well plate with a cell density of 3.5*10 4 cells / mL, 100 μl / well, when the cell confluency reaches 60-70%, replace with serum-free culture, and infect with adenovirus; the above-mentioned adenoviruses are the overexpressed HIP-55 recombinant adenovirus prepared in Example 1 respectively ( Dsred-HIP55), Des1d adenovirus (Dsred), shRNA interference HIP-55 expression recombinant adenovirus (KD1), control shRNA interference recombinant adenovirus (scramble), pAd / PL-Dest adenovirus. After culturing for 24 hours, give isoproterenol (ISO, the final concentration is 10 -5 mol / L) were collected after 24 hours of treatment. Six hours before sample collection, 10 μl / well of BrdU was added (final concentration 10 μmol / L). Discard the supernatant when collecting s...

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Abstract

The invention discloses RNA and application of RNA in diseases of a cardiovascular system. According to RNA, the nucleotide sequence of RNA is a sequence 3 in a sequence table. The invention further provides a gene,encoding RNA. The nucleotide sequence of the gene of RNA is a sequence 4 in the sequence table. The experiment of the invention shows that RNA and the application of RNA in the diseases of the cardiovascular system find that by an induction of isoproterenol (ISO), RNAi method Knock down IHP-55 is used in neonatal rat cardiac fibroblasts to obviously promote proliferation of cardiac fibroblasts; ISO induced activation and kinase activity of P38MAPK can be enhanced in the Knock-down HIP-55 cardiac fibroblasts; and HIP-55 is a new protein which has a protective effect on cardiac fibrosis, and provides a new target for clinical diagnosis, treatment and new medicament development.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an RNA and its application in cardiovascular system diseases. Background technique [0002] HIP-55 (hematopoietic progenitor kinase 1 [HPK1]-interacting protein of 55 kDa; also known as drebrin-like protein, mAbp1 or SH3P7) is a protein containing a multifunctional domain. Its structure mainly includes the N-terminal F-actin protein binding domain and the C-terminal SH3 domain. HIP-55 is involved in synaptogenesis, receptor endocytosis, cytoskeleton rearrangement and signal transduction. At present, little is known about the function of HIP-55, and there is no research report on the role of HIP-55 in the cardiovascular system. [0003] Cardiovascular disease is the "number one killer" of Chinese residents' health, and the annual direct medical expenses mainly for cardiovascular disease have reached 130 billion yuan. Myocardial fibrosis is an important pathological change of cardia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/861C12N5/10C12N7/01A61K48/00A61K45/00A61P9/00
Inventor 李子健张幼怡邢瑞杨承志吕志珍
Owner PEKING UNIV THIRD HOSPITAL
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