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Function of IRF (Interferon Regulatory Factor) 3 gene in atherosclerosis and application of inhibitor of IRF3 gene

A technology of atherosclerosis and inhibitors, applied in the field of gene function and application, can solve problems such as unsatisfactory treatment and control effects, and achieve the effect of worsening atherosclerosis

Active Publication Date: 2014-05-14
武汉惠康达科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many risk factors for atherosclerosis that have been discovered so far, but the relevant targeted treatment and control effects are not satisfactory. Lipid lowering and anti-inflammatory treatment are currently the most important treatment measures

Method used

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  • Function of IRF (Interferon Regulatory Factor) 3 gene in atherosclerosis and application of inhibitor of IRF3 gene
  • Function of IRF (Interferon Regulatory Factor) 3 gene in atherosclerosis and application of inhibitor of IRF3 gene
  • Function of IRF (Interferon Regulatory Factor) 3 gene in atherosclerosis and application of inhibitor of IRF3 gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Mouse Atherosclerosis Model (AS) Obtained

[0031] 1. Grouping of experimental animals: 8 weeks old, weighing 19-25g, male, ApoE - / - mouse and IRF3 - / - ApoE - / - Mice were fed high-fat diet (Western Diets, HFD) respectively, ApoE - / - HFD group, IRF3 - / - ApoE - / - There are 2 groups in the HFD group.

[0032] 2. Atherosclerosis model induced by high-fat diet:

[0033] Using ApoE - / - mouse and IRF3 - / - ApoE - / -In mice, the AS model was established, and the phenotype correlation analysis was carried out to clarify that the IRF3 gene plays an important role in the pathogenesis of atherosclerosis. Mice were fed a high-fat diet from 8 weeks of age until sacrificed at 28 weeks and samples were collected.

Embodiment 2

[0034] Example 2 Determination of plaque area in AS model mice

[0035] 1. Final mouse tissue harvesting

[0036] Feed the mice with high-fat diet until 28 weeks, weigh them, anesthetize the mice with 3% pentobarbital sodium, 90 mg / kg, fix them on the sampling board with a needle, moisten the skin of the chest and abdomen of the mice with gauze, and cut them with ophthalmic scissors. Open the thorax, expose the heart, cut the right atrial appendage, insert the needle of the infusion set into the left ventricle, slowly inject 10-15mL PBS buffer solution with a 50mL syringe, wait until the effluent from the right atrial appendage is clear, replace it with 4% paraformaldehyde and continue pushing Inject 10-15mL. After the perfusion, the thoracoabdominal viscera were removed and only the heart was kept. Put the mouse under a microscope, separate the fascia and adipose tissue around the aortic arch, cut off the brachiocephalic trunk, put it into a 5mL EP tube filled with 4% paraf...

Embodiment 3

[0048] Example 3 Analysis of Plaque Contents in AS Model Mice

[0049] 1. Macrophage and Smooth Muscle Cell Expression Assay

[0050] Immunofluorescent staining was used to detect the expression of macrophages (CD68) and smooth muscle actin (SMA). Required primary antibody information: CD68 (MCA1957; 1:100; rat; AbD Serotec), SMA (ab5694; 1:100; rabbit; Abcam); Required secondary antibody information: Alexa Flour? 568 goat anti-rat IgG (A11077 ; Invitrogen, Carlsbad, CA), Alexa Fluor 488-conjugated goat anti-rabbit IgG (A11008; Invitrogen, Carlsbad, CA).

[0051] The main steps are:

[0052] 1) Baked slices: put the paraffin slices in the oven for more than 30 minutes.

[0053] 2) Dewaxing: xylene 5min x 3 times.

[0054] 3) Hydration: 100% ethanol for 5 min×2 times; 95% ethanol for 5 min; 70% ethanol for 5 min; ddH 2 O soaking for 5min × 2 times.

[0055] 4) Citrate tissue antigen repair (high pressure repair): Take a certain amount of pH6.0 citrate antigen repair worki...

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Abstract

The invention discloses the function of an IRF3 gene in atherosclerosis and an application of an inhibitor of the IRF3 gene, belonging to the field of the function and the application of a gene. According to the invention, ApoE- / -mice and IRF3- / -ApoE- / -mice are taken as experimental subjects, an atherosclerosis model is obtained through high-fat induction, AS model mouse plaque area measurement and inclusion analysis are performed, and the results show that the aorta plaque area of the IRF3- / -ApoE- / -mice is remarkably reduced in comparison with that of the ApoE- / -mice. The invention discloses the function of the IRF3 gene in an atherosclerosis disease, which means that the IRF3 gene has an effect of promoting the formation of the aorta plaques, especially that the IRF3 gene can worsen the atherosclerosis. According to the abovementioned function, the IRF3 can be used as a drug target for screening drugs for treating the atherosclerosis disease. The inhibitor of the IRF3 can be used for preparing the drug for treating the atherosclerosis disease.

Description

[0001] technical field [0002] The invention belongs to the field of gene function and application, and relates to the function of IRF3 (interferon regulatory factor 3) gene in atherosclerosis and the application of its inhibitor. Background technique [0003] Cardiovascular and cerebrovascular diseases are the main cause of death in many developed countries, and the morbidity and mortality in my country are also increasing year by year. The basis of cardiovascular and cerebrovascular diseases is atherosclerosis (Atheosclersis, AS). Atherosclerosis can thicken and harden the arterial wall and narrow the lumen, leading to many cardiovascular and cerebrovascular events. Acute stenosis and occlusion of coronary arteries caused by rupture of atherosclerotic unstable plaques, platelet aggregation, and thrombus formation are important causes of acute coronary syndrome (ACS). [0004] Atherosclerosis is a chronic inflammatory disease involving multiple genetic genes, risk factors...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K45/00A61K39/395A61P9/10
Inventor 李红良蒋曦蒋丁胜胡俊飞邓克穷王丕晓
Owner 武汉惠康达科技有限公司
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