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Small numerator interference RNA for inhibiting vascellum endometrial hyperplasia

A technology of small molecule interference and vascular intima, applied in DNA/RNA fragments, cardiovascular system diseases, recombinant DNA technology, etc.

Inactive Publication Date: 2010-09-15
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, recently published results of randomized trials suggest that DES can increase the risk of long-term non-cardiac death and cardiac death and / or myocardial infarction caused by late thrombosis (Pfisterer M, Brunner-La Rocca HP, Buser PT, Rickenbacher P , Hunziker P, Mueller C, Jeger R, Bader F, Osswald S, Kaiser C; BASKET-LATE Investigators. Late clinical events after clopidogrel discontinuation may limit the benefit of drug-eluting stents: an observational study of drug-eluting versus bare-metal stents . J Am Coll Cardiol. 2006 Dec 19; 48(12): 2584-91)

Method used

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  • Small numerator interference RNA for inhibiting vascellum endometrial hyperplasia
  • Small numerator interference RNA for inhibiting vascellum endometrial hyperplasia
  • Small numerator interference RNA for inhibiting vascellum endometrial hyperplasia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1, ADAMTS-7siRNA inhibits ADAMTS-7 in primary cultured smooth muscle cells

[0029] Firstly, ADAMTS-7-specific small interfering RNA was designed and synthesized. The specific method is as follows: according to the rat ADAMTS-7 sequence (GenBank Accession Number NM_001047101) by Invitrogen Company Blcok-iT TM RNAi Designer software design ADAMTS-7siRNA sequence: the sense strand of ADAMTS-7siRNA is: 5'-UAAUAAGGCGCACAACGGUGAUGUG-3' (sequence 1 in the sequence table), the antisense strand of ADAMTS-7siRNA is: 5'-UAAUAAGGCGCACAACGGUGAUGUG-3' (sequence Sequence 2) in the list. ADAMTS-7 siRNA of the above sequence was synthesized.

[0030]50nmol / L ADAMTS-7siRNA was transfected with Oligofectamine (purchased from Invitrogen, CA) into primary cultured rat aortic smooth muscle cells (primary rat aortic smooth muscle cells were prepared by conventional tissue patch method), 48 After 1 hour, the RNA of the cells was extracted, reverse-transcribed into cDNA and used as...

Embodiment 2

[0033] Example 2, ADAMTS-7siRNA inhibits the migration of primary cultured smooth muscle cells

[0034] Using the classic scratch test of isolated smooth muscle cells to simulate vascular injury, to prove that ADAMTS-7siRNA can inhibit the migration of smooth muscle cells, the specific experiments are as follows:

[0035] 50nmol / L ADAMTS-7siRNA constructed in the above-mentioned Example 1 was transfected into primary cultured mouse smooth muscle cells with Oligofectamine (purchased from Invitrogen, CA), and the primary cultured smooth muscle cells transfected with Oligofectamine and transfected Primary cultured smooth muscle cells transfected with scramble siRNA were used as control. Scratch after 48 hours, observe and take pictures after scratching 6 hours, and compare the relative distance of cell migration. The experiment was repeated three times. The images of cell migration in each group are as follows figure 2 A (N=3, P figure 2 Shown in B. Among them, Vehicle is th...

Embodiment 3

[0036] Example 3, ADAMTS-7siRNA inhibits the level of ADAMTS-7 protein in blood vessels after rat carotid artery strain

[0037] 1. Establishment of balloon strain injury model in rats

[0038] 200-220 grams of male Sprague-Dawley rats were intraperitoneally injected with chloral hydrate (300 mg / kg). A 1.5mm balloon (Medtronic, Minneapolis, MN) was inserted into the common carotid artery from the left external carotid artery and advanced 4 cm toward the thoracic aorta. This process was repeated 3 times to ensure complete removal of the endothelium, and similar tissue separation was performed on the contralateral vessel. At the same time, rats without balloon strain (indicated by Sham) were used as controls.

[0039] The common carotid arteries were fixed and harvested 7 days and 14 days after modeling, and identified by histological staining. The result is as Figure 4 shown. Among them, Sham means the rats in the control group, and Injury means the rats in the balloon st...

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Abstract

The invention discloses small interfering RNA for suppressing vascular intimal hyperplasia. The nucleotide sequence of sense chain of the small interfering RNA is represented by sequence 1 in the sequence table, and the nucleotide sequence of antisense chain of the small interfering RNA is represented by sequence 2 in the sequence table. The small interfering RNA for suppressing vascular intimal hyperplasia can be directly applied to vascular adventitia to persistently and remarkably suppress vascular intimal hyperplasia, and can be used for treating restenosis after angioplasty.

Description

technical field [0001] The invention relates to a small molecular interference RNA for inhibiting vascular intimal hyperplasia. Background technique [0002] RNA interference (RNA Interference, RNAi) refers to the phenomenon of specifically inhibiting the post-transcriptional expression of target genes due to the introduction of exogenous or endogenous double-stranded RNA into cells. Since its discovery in 1998, as an efficient and specific technology for regulating gene expression, RNAi has become a powerful tool for gene function research. In just a few years, the research of RNAi has made rapid development, such as the first report in 2001 that the successful application of RNAi technology in mammalian cells to inhibit gene expression (Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans Nature. 1998 Feb 19; 391(6669): 806-11.), pioneered the application of RNAi technology t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A61K31/713A61K48/00A61K47/34A61P9/00A61K47/36C12N15/113
Inventor 王宪孔炜王利
Owner PEKING UNIV
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