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Function and application of IRF5 (interferon regulatory factor-5) and IRF5 inhibitor in treatment of restenosis after VI (vascular injury)

A technology for vascular injury and restenosis, applied in gene therapy, cardiovascular system diseases, blood diseases, etc., can solve problems such as high incidence and affecting treatment effect

Inactive Publication Date: 2017-02-15
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is currently no cure for this type of disease. The main treatment of vascular surgery is vascular reconstruction of the occluded segment, including balloon dilation, stent placement, and arterial bypass surgery. Although vascular reconstruction can be achieved and the blocked artery can be effectively dredged, Improve blood supply, but the incidence of restenosis after vascular reconstruction is high (30-60%), which greatly affects the treatment effect. So far, restenosis after vascular reconstruction is still a clinical problem

Method used

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  • Function and application of IRF5 (interferon regulatory factor-5) and IRF5 inhibitor in treatment of restenosis after VI (vascular injury)
  • Function and application of IRF5 (interferon regulatory factor-5) and IRF5 inhibitor in treatment of restenosis after VI (vascular injury)
  • Function and application of IRF5 (interferon regulatory factor-5) and IRF5 inhibitor in treatment of restenosis after VI (vascular injury)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] [Example 1] Mouse vascular injury model (VI) obtained

[0045] 1. Grouping of experimental animals: WT and IRF5-KO mice aged 8-10 weeks and weighing 24-27 g were used, and were divided into two groups: WT vascular injury group and IRF5-KO vascular injury group, with 20 mice in each group. mouse. The mice were sacrificed 14 days and 28 days after the operation, and blood vessels in the injured segment were collected for analysis.

[0046] 2. Operation procedure of mouse vascular injury model:

[0047] 1) Accurately weigh the body weight (g) of the mouse in dynamic mode with an electronic balance, accurately prepare 3% pentobarbital sodium solution with double distilled water, shake gently to dissolve it fully, and use 80 mg / kg body weight dose to calculate Accurately extract the corresponding volume of solution with a 1mL syringe after the required volume of pentobarbital sodium solution, and perform intraperitoneal injection to anesthetize the mouse. After the mouse i...

Embodiment 2

[0052] [Example 2] Intimal Neonatal Determination in Vascular Injury Model (VI) Mice

[0053] 1. Mice collection

[0054] 1) Anesthetize the mouse, cut the heart and let the blood out.

[0055] 2) Cut the carotid artery from the proximal bifurcation of the carotid artery, take 0.5-0.6cm long, and keep the external carotid artery knot.

[0056] 3) Put the carotid artery into PBS, and gently drain the residual blood in the lumen with micro forceps.

[0057] 4) Put the blood vessel into a 1.5mL EP tube filled with 1mL 4% paraformaldehyde for fixation.

[0058] 2. Pathological detection

[0059] 2. Pathological detection

[0060] 2.1 Preparation of paraffin specimen slices

[0061] Paraffin specimen slices are prepared by professional pathologists in the laboratory. The main operating procedures include: after overnight fixation in 4% paraformaldehyde, carefully wrap the blood vessel with filter paper, put it into the embedding frame→rinse with running water→dehydration→trans...

Embodiment 3

[0069] [Example 3] Detection of the proliferation level of blood vessel wall cells

[0070] Immunofluorescent staining was used to detect the expression of proliferating cell nuclear antigen (PCNA) and cell cycle protein (Cyclin D1). Required primary antibody information: PCNA (#2586; 1:100; mouse; Cell Signaling Technology), cyclin D1 (#2978; 1:25; rabbit; Cell Signaling Technology); required secondary antibody information: Alexa Fluor 568-conjugated goat anti-rabbit IgG (A11011; Invitrogen, Carlsbad, CA), Alexa Fluor 568-conjugated goat anti-mouse IgG (A11004; Invitrogen, Carlsbad, 150d, CA).

[0071] The main steps are:

[0072] 1) Baked slices: put the paraffin slices in an oven at 55°C for more than 60 minutes.

[0073] 2) Dewaxing: xylene 8min×3.

[0074] 3) Hydration: 100% ethanol 5min×2; 95% ethanol 5min; 70% ethanol 5min; ddH 2 O dipping for 5min×2.

[0075] 4) Citrate tissue antigen repair (high pressure repair): Take a certain amount of pH6.0 citrate antigen repa...

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Abstract

The invention discloses a function and an application of an IRF5 (interferon regulatory factor-5) and an IRF5 inhibitor in treatment of restenosis after VI (vascular injury). An IRF5 gene knockout mouse and a wild mouse are taken as experimental subjects, detection of intima neogenesis, vessel wall cell proliferation level and smooth muscle cell phenotype transformation of the mice are performed through a VI model, and results show that IRF5 gene knockout can obviously inhibit intima neogenesis and cell proliferation and inhibit transformation of smooth muscle cells from contractile phenotype to synthetic phenotype. Therefore, the function of the IRF5 on treatment of restenosis after VI mainly lies in promoting intima neogenesis, cell proliferation and smooth muscle cell phenotype transformation. Based on the function of the IRF5, the IRF5 can be taken as a drug target to be used for screening drugs for preventing and treating restenosis after VI, and the IRF5 inhibitor can be used for preparing a drug and an arterial stent for preventing and treating restenosis after VI.

Description

technical field [0001] The invention belongs to the field of gene function and application, and particularly relates to the application of an interferon regulatory factor-5 (interferonregulatory factor-5, IRF5) as a drug target in screening drugs for restenosis after blood vessel injury. Background technique [0002] With the rapid development of human society and economy, the improvement of people's living standards, the change of diet structure and the aging process of the population, the incidence of cardiovascular disease is increasing year by year, and it has become one of the major diseases that seriously endanger the global public health. Neointimal formation is a common pathological process of angioproliferative vascular diseases such as atherosclerosis (AS), pulmonary hypertension, and restenosis after percutaneous transluminal coronary intervention (PCI). The main pathological basis of restenosis is the hyperplasia of neointima and media and the accompanying format...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K48/00A61K31/713A61K45/00A61L31/16A61P7/00A61P9/10A61P9/12
CPCA61K49/0008A61K31/713A61K45/00A61K48/005A61L31/16A61L2300/432
Inventor 李红良程文林秦娟娟
Owner WUHAN UNIV
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