The function and application of irf9 gene in atherosclerosis
A technology for atherosclerosis and aorta, which is 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 atherosclerotic diseases
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Embodiment 1
[0026] Example 1 Mouse Atherosclerosis Model (AS) Obtained
[0027] 1. Grouping of experimental animals: 8 weeks old, weighing 19-25g, male, LDLR - / - Mice and IRF9 - / - LDLR - / - Mice were fed 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.
[0028] 2. Atherosclerosis model induced by high-fat diet:
[0029] Using LDLR - / - Mice and IRF9 - / - LDLR - / - In mice, the AS model was established, and the phenotype correlation analysis was carried out to clarify that the IRF9 gene plays an important role in the pathogenesis of atherosclerosis. Mice were fed feed from 8 weeks of age until sacrificed at 28 weeks and samples were collected.
Embodiment 2
[0030] Example 2 Determination of plaque area in AS model mice
[0031]1. Final mouse tissue harvesting
[0032] 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% parafo...
Embodiment 3
[0044] Example 3 Analysis of Plaque Contents in AS Model Mice
[0045] 1. Macrophage and Smooth Muscle Cell Expression Assay
[0046] 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).
[0047] The main steps are:
[0048] 1) Baked slices: put the paraffin slices in the oven for more than 30 minutes.
[0049] 2) Dewaxing: xylene 5min x 3 times.
[0050] 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.
[0051] 4) Citrate tissue antigen repair (high pressure repair): Take a certain amount of pH6.0 citrate antigen repair worki...
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