Novel use of Kruppel-like transcription factor 4

A technology of transcription factors and coding genes, applied in gene therapy, pharmaceutical formulations, drug combinations, etc., can solve the problems of death, reduce the incidence of postoperative restenosis, etc., and achieve the effect of inhibiting proliferation, obvious inhibitory effect, and inhibiting hyperplasia

Inactive Publication Date: 2009-08-12
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The emergence of intravascular drug-eluting stents (DES) has solved the problem of acute occlusion caused by elastic retraction of blood vessels, and at the same time inhibited excessive proliferation of vascular intima by inhibiting cell proliferation, significantly reducing the incidence of postoperative restenosis, but Long-term use of stents can lead to intravascular thrombosis leading to myocardial infarction and cardiac death

Method used

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  • Novel use of Kruppel-like transcription factor 4
  • Novel use of Kruppel-like transcription factor 4
  • Novel use of Kruppel-like transcription factor 4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, the construction of recombinant adenovirus

[0032] 1. Construction method of recombinant adenovirus AdKLF4:

[0033] Plasmid pMT-KLF4 (Janiel M. Shields, Robert J. Christy and Vincent W. Identification and Characterization of a Gene Encoding a Gut-enriched Krüppel-like Factor Expressed during Growth Arrest. J Biol Chem. 1996 Aug16; 271(33): 20009 -17) (Peking University);

[0034] Shuttle plasmid pAdlox containing tetracycline response enhancer and CMV promoter (Nanping Wang, Lynne Verna, Stephen Hardy, John Forsayeth, Yi Zhu and Michael B. Stemerman. Adenovirus-Mediated Overexpression of c-Jun and c-Fos Induces Intercellular Adhesion Molecule-1 and Monocyte Chemoattractant Protein-1 in Human Endothelial Cells.Arterioscler.Thromb.Vasc.Biol.1999; 19; 2078-2084) (Peking University);

[0035] Delete the adenovirus 5 genomic DNA containing the loxp site in the E1 and E3 regions (Sauer, B.1987.Functional expression of the cre-lox site-specific recombination ...

Embodiment 2

[0057] Example 2, Inhibition of KLF4 on the proliferation of vascular smooth muscle cells

[0058] 1. Isolation and primary culture of rat thoracic aortic smooth muscle cells

[0059] 1. Isolation method: the isolated SD male rat thoracic aorta was used as the raw material, and the rat aortic smooth muscle cells were isolated by the tissue patch method.

[0060] Verification: α-actin is a marker of vascular smooth muscle cells, and the above-mentioned isolated cells were identified by immunostaining with an antibody specific to α-actin (purchased from Santa Cruz Biotechnology, Inc, Cat. No. sc-130617) , the results showed that the positive cells were more than 99%, which proved that the isolated cells were vascular smooth muscle cells.

[0061] Primary culture: use DMEM medium containing 10% (volume percentage) fetal bovine serum, 100U / mL penicillin, 100mg / mL streptomycin, at 37°C, 5% CO 2 cultured in an incubator.

[0062] 2. Construction of recombinant smooth muscle cells...

Embodiment 3

[0078] Example 3, KLF4 inhibits the hyperplasia of vascular intima after rat carotid artery balloon injury

[0079] 1. Treatment of rat carotid artery balloon injury:

[0080] 400 g male Sprague-Dawley rats were anesthetized by intraperitoneal injection of 0.6% sodium pentobarbital (injection dose: 25 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. After the balloon was inflated (0.5 atmospheres), it was rotated and pulled back to the bifurcation of the common carotid artery , this process was repeated 3 times to ensure that the vascular endothelium was fully removed. At the same time, rats without balloon strain (indicated by control) were used as a control.

[0081] The total RNA of blood vessels was extracted 3 days after the balloon injury, and the changes of KLF4mRNA level in injured blood vessels were detected by RT-PCR. The primers used i...

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Abstract

The invention discloses new application of KFL4, a kruppel-like transcription factor. The application refers to the application of coding gene of the kruppel-like transcription factor 4 protein in preparing medicines for treating intimal hyperplasia. The invention discloses a medicine with the kruppel-like transcription factor 4 protein or coding gene thereof as target spots for treating intimal hyperplasia, the active ingredients thereof are micro RNA or si RNA or recombinant vector of coding gene containing the kruppel-like transcription factor 4 protein, which overexpress the kruppel-like transcription factor 4. Experiments indicate that the application method or the medicine of the invention can effectively inhibit proliferation of vascular smooth muscle cell, so as to inhibit hyperplasia of endangium, the inhibiting effect is very obvious; therefore, the method and medicine of the invention enjoy very broad application prospect in treating hyperplasia of endangium.

Description

technical field [0001] The present invention relates to a novel use of Krüppel-like transcription factor 4 (KLF4). Background technique [0002] Percutaneous coronary angioplasty (PTCA) is an important method for the treatment of coronary heart disease, but postoperative vascular restenosis has always been the main problem affecting the efficacy of PTCA. The big disadvantage is that the incidence of restenosis can reach 30% to 50% within six months after surgery. The emergence of intravascular drug-eluting stents (DES) has solved the problem of acute occlusion caused by elastic recoil of blood vessels, and at the same time inhibited excessive proliferation of vascular intima by inhibiting cell proliferation, significantly reducing the incidence of postoperative restenosis, but Long-term use of stents can lead to intravascular thrombosis, resulting in myocardial infarction and cardiac death. To develop more specific drugs to inhibit the formation of neointima is a very nece...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P9/00
Inventor 汪南平王颖
Owner PEKING UNIV
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