Function and application of interferon regulatory factor 9 (IRF9) gene in atherosclerosis

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

Active Publication Date: 2014-05-14
武汉惠康基因科技有限公司
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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 and application of interferon regulatory factor 9 (IRF9) gene in atherosclerosis
  • Function and application of interferon regulatory factor 9 (IRF9) gene in atherosclerosis
  • Function and application of interferon regulatory factor 9 (IRF9) gene in atherosclerosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Obtaining a mouse atherosclerosis model (AS)

[0029] 1. Grouping of experimental animals: 8-week-old, weighing 19-25g, male, LDLR - / - Mice and IRF9 - / - LDLR - / - Mice, fed with high-fat diet (Western Diets, HFD) and low-fat diet (Normal chow, NC), respectively, LDLR - / - HFD group, LDLR - / - NC group, IRF9 - / - LDLR - / - HFD group, IRF9 - / - LDLR - / - There are 4 groups in the NC group.

[0030] 2. The atherosclerosis model is induced by high-fat diet. The operation process:

[0031] LDLR - / - Mice and IRF9 - / - LDLR - / -In mice, an AS model was established, and phenotype correlation analysis was performed to clarify that IRF9 gene plays an important role in the pathogenesis of atherosclerosis. Mice were fed chow from 8 weeks of age until sacrificed at 28 weeks and samples were collected.

Embodiment 2

[0032] Example 2 Determination of plaque area in AS model mice

[0033] 1. Mice terminal tissue harvesting

[0034] Mice were fed high-fat diet until 28 weeks, weighed, anesthetized the mice with 3% sodium pentobarbital, 90 mg / kg, fixed on the sampling plate with a needle, moistened the skin of the chest and abdomen of the mice with gauze, and cut them with ophthalmic scissors. Open the thoracic cavity, expose the heart, cut the right atrial appendage, insert the needle of the infusion set into the left ventricle, slowly inject 10-15 mL of PBS buffer with a 50 mL syringe, and when the effluent from the right atrial appendage is clear, replace it with 4% paraformaldehyde and continue to push Note 10-15mL. After the perfusion, the thoracic and abdominal organs were removed, and only the heart was preserved. The mice were placed under a microscope, the fascia and adipose tissue around the aortic arch were separated, the brachiocephalic trunk was cut off, and placed in a 5 mL EP...

Embodiment 3

[0046] Example 3 Analysis of plaque inclusions in AS model mice

[0047] 1. Macrophage and Smooth Muscle Cell Expression Assay

[0048] Immunofluorescence staining was used to detect the expression of macrophages (CD68) and smooth muscle actin (SMA). Primary Antibody Information Required: CD68 (MCA1957; 1:100; rat; AbD Serotec), SMA (ab5694; 1:100; rabbit; Abcam); Secondary Antibody Information Required: Alexa Flour? 568 goat anti-rat IgG (A11077 ; Invitrogen, Carlsbad, CA), Alexa Fluor 488-conjugated goat anti-rabbit IgG (A11008; Invitrogen, Carlsbad, CA).

[0049] The main steps are:

[0050] 1) Baked slices: Place the paraffin slices in the oven for more than 30 minutes.

[0051] 2) Dewaxing: xylene 5min×3 times.

[0052] 3) Hydration: 100% ethanol 5min×2 times; 95% ethanol 5min; 70% ethanol 5min; ddH 2 O dipping 5min×2 times.

[0053] 4) Citrate tissue antigen retrieval (high pressure repair): Take a certain amount of pH6.0 citrate antigen retrieval working solution ...

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Abstract

The invention discloses a function and application of an interferon regulatory factor 9 (IRF9) gene in atherosclerosis (AS), belonging to the field of gene functions and applications. According to the function and application disclosed by the invention, LDLR (Low Density Lipoprotein Receptor)-/- mice and IRF9-/-LDLR-/- mice are taken as experimental subjects, an atherosclerosis model is obtained by high fat induction, and AS model mouse plaque area determination and inclusion analysis are carried out; results show that compared with LDLR-/- mice, the aortal plaque area of the IRF9-/-LDLR-/- mice is significantly increased. The results indicate that the function of IRF9 gene in atherosclerosis is manifested by the action in inhibiting aortal plaque formation, especially inhibiting atherosclerosis. As for the function of the IRF9 gene, IRF9 gene can be used for preparing medicaments for treating atherosclerosis.

Description

[0001] Technical field [0002] The invention belongs to the function and application of genes, involving the function and application of an IRF9 (Interfereon Regulatory Factor 9) gene in atherosclerosis. Background technique [0003] Cardiovascular and cerebrovascular diseases are the main causes of death in many developed countries, and the incidence and mortality in my country have also increased year by year.The basis of cardiovascular and cerebrovascular diseases is atherosclerosis (AS). Atherosclerosis can increase the wall of the arteries, hardening, and narrow lumen, resulting in many cardiovascular and cerebrovascular events.The rupture of atherosclerotic unstable plaques, the aggregation of platelets, and the acute stenosis and occlusion of the coronary arteries caused by thrombosis are important causes of acute coronary syndrome (ACS). [0004] Atherosclerosis is a chronic inflammatory disease jointly participated in a variety of genetic genes, risk factors and immune ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/17A61P9/10
Inventor 李红良黄玲向梅蒋曦王丕晓张晓东
Owner 武汉惠康基因科技有限公司
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