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Function of IRF (Interferon Regulatory Factor) 9 in stent and carotid endarterectomy restenosis as well as application of inhibitor of IRF9

A technology of restenosis and inhibitor, applied in the application field of function and its inhibitor, can solve the problems of high incidence and affect the treatment effect, and achieve the effect of promoting restenosis

Active Publication Date: 2014-05-14
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Currently, there is no cure for this kind of disease. The main treatment of vascular surgery is vascular reconstruction of occluded segment, including balloon dilation, stent placement, and arterial bypass surgery, etc. However, the incidence of restenosis after vascular reconstruction is high (30 % to 60%), which greatly affected the therapeutic effect, so far restenosis after vascular reconstruction is still a clinical problem

Method used

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  • Function of IRF (Interferon Regulatory Factor) 9 in stent and carotid endarterectomy restenosis as well as application of inhibitor of IRF9
  • Function of IRF (Interferon Regulatory Factor) 9 in stent and carotid endarterectomy restenosis as well as application of inhibitor of IRF9
  • Function of IRF (Interferon Regulatory Factor) 9 in stent and carotid endarterectomy restenosis as well as application of inhibitor of IRF9

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Mouse Vascular Injury Model (VI) Obtained

[0033] 1. Grouping of experimental animals: WT and IRF9-KO mice aged 8-10 weeks and weighing 24-27g were used and divided into four groups: WT vascular injury group; WT sham operation group; IRF9-KO vascular injury group; IRF9 - KO sham operation group, 60 mice in each group. Twenty mice in each group were sacrificed 7 days, 14 days, and 28 days after the operation, and blood vessels in the injured segment were collected for analysis.

[0034] 2. Operation procedure of mouse vascular injury model:

[0035] 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 80mg / kg body weight dose to calculate Accurately extract the corresponding volume of pentobarbital sodium solution with a 1mL syringe, and intraperitoneally inject the anesthetized mouse. After t...

Embodiment 2

[0040] Example 2 Determination of Intimal Neogenesis in Vascular Injury Model (VI) Mice

[0041] 1. Mice Harvesting

[0042] 1) Anesthetize the mouse, cut the heart and let the blood out.

[0043] 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.

[0044] 3) Put the carotid artery in PBS, and gently drain the residual blood in the lumen with micro forceps.

[0045] 4) Put the blood vessel into a 1.5mL EP tube filled with 1mL 4% paraformaldehyde for fixation.

[0046] 2. Pathological detection

[0047] 2.1 Preparation of paraffin specimen slices

[0048] Paraffin specimen sections are prepared by professional pathological staff in the laboratory. The main operating procedures include trimming the heart → processing the embedding frame → washing with running water → dehydrating → transparent → dipping in wax → embedding → sectioning (3 μm) → spreading → drying or baking Backup.

[...

Embodiment 3

[0056] Example 3 Detection of proliferation level of blood vessel wall cells

[0057] The expressions of proliferating cell nuclear antigen (PCNA) and cell cycle protein (Cyclin D1) were detected by immunofluorescence staining. 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, 150 d, CA).

[0058] The main steps are:

[0059] 1) Baked slices: put the paraffin slices in the oven for more than 30 minutes.

[0060] 2) Dewaxing: Xylene 5min×3.

[0061] 3) Hydration: 100% ethanol 5min×2; 95% ethanol 5min; 70% ethanol 5min; ddH 2 O dipping for 5min×2.

[0062] 4) Citrate tissue antigen repair (high pressure repair): Take a certain amount of pH6.0 citrate antigen repair working...

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Abstract

The invention discloses the function of IRF 9 in stent and carotid endarterectomy restenosis as well as an application of an inhibitor of IRF9, belonging to the field of the function and the application of a gene. According to the invention, IRF9 gene knockout mice and wild C57 mice are taken as experimental subjects, detection of mouse intima neogenesis, vascular wall cell proliferation level and smooth muscle cell phenotype is performed through a vascular injury model, and the results show that IRF9 gene knockout mice represents inhibited intima neogenesis and cell proliferation in comparison with the wild C57 mice. The invention discloses the function of the IRF9 gene in the stent and carotid endarterectomy restenosis, which mainly shows that the IRF9 gene has the function of promoting the intima neogenesis and cell proliferation. According to the abovementioned function, the IRF9 can be used as a drug target for screening drugs for treating hemadostenosis diseases, and the inhibitor of the IRF9 can be used for preparing the drug for treating the stent and carotid endarterectomy restenosis.

Description

[0001] technical field [0002] The invention belongs to the field of gene function and application, and relates to the function of IRF9 in restenosis after stent and endarterectomy and the application of its inhibitor. Background technique [0003] In China at present, with the change of diet structure and the aging process of the population, atherosclerotic obliterative lesions show a trend of increasing year by year and become one of the main causes of death in our country. At present, there is no cure for this kind of disease. The treatment methods of vascular surgery include balloon dilation, stent placement and arterial bypass, but restenosis after vascular reconstruction greatly affects the treatment effect. Research on vascular restenosis has been carried out for many years, but so far it is not clear. Studies have shown that in the process of injury formation, the excessive proliferation of neointima and media tissue and the accompanying formation of extracellular ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K48/00A61K39/395A61P9/10
Inventor 李红良张书敏朱丽华张晓东蒋丁胜向梅
Owner WUHAN UNIV
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