Application of Mindin gene in treatment of atherosclerosis
A technology of atherosclerosis and genes, which is applied in the field of gene functions and applications, can solve the problems of unexplained biological functions and possible mechanisms of action, and achieve the effect of promoting atherosclerosis
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Embodiment 1
[0038] Example 1 Mouse Atherosclerosis Model (AS) Obtained
[0039] 1. Grouping of experimental animals: male ApoE- / - mice and Mindin- / -ApoE- / - mice aged 8 weeks, weighing 19-25g, were fed with high-fat diet (Western Diets, HFD) respectively, ApoE - / - HFD group and Mindin- / -ApoE- / - HFD group consisted of 2 groups, with 20 rats in each group.
[0040] 2. Atherosclerosis model induced by high-fat diet:
[0041] ApoE- / - mice and Mindin - / -ApoE- / - mice were used to establish AS models, and phenotype correlation analysis was performed to clarify the role of Mindin gene in atherosclerotic diseases. From the age of 8 weeks, the mice in the HFD group were fed with high-fat diet for 28 weeks and sacrificed, and the samples were collected.
Embodiment 2
[0042] Example 2 Determination of plaque area in AS model mice
[0043] 1. Final mouse tissue harvesting
[0044] Mice were fed with high-fat or low-fat diet until 28 weeks, weighed, anesthetized with 3% pentobarbital sodium, 90 mg / kg, fixed on the sampling board with a needle, moistened the skin of the chest and abdomen of the mouse with gauze, and used Ophthalmic scissors cut open the chest cavity, expose the heart, cut open the right atrial appendage, insert the needle of the infusion set into the left ventricle, inject 10-15mL PBS buffer slowly with a 50mL syringe, wait until the right atrial appendage effluent is clear, replace it with 4% Polymer Formaldehyde continued to 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% paraformaldehyde, cut ...
Embodiment 3
[0057] Example 3 Determination of Plaque Stability in AS Model Mice
[0058] 1. Determination of collagen content in aortic sinus:
[0059] Sirius Red (PSR) staining, the main steps are: bake at 55°C for 30 minutes → xylene for 2 minutes, 3 times → 100% alcohol for 1 minute → 95% alcohol for 1 minute → 70% alcohol for 1 minute → rinse with running water for 10 minutes → double distilled water for 1 minute → mass fraction 0.2% phosphomolybdic acid for 2 minutes → 0.1% Sirius scarlet picric acid solution was dropped on the tissue, stained in a wet box for 90 minutes → remove the residue → 0.01N hydrochloric acid for 4 seconds → 70% alcohol once → 90% alcohol once → 100% alcohol 30s, 3 times → 2min in xylene, 3 times → cover the slide immediately while the xylene is still wet, and take pictures under the microscope.
[0060] Collagen ratio (%)=collagen area / plaque area*100%.
[0061] 2. Expression Determination of Aortic Sinus Macrophages and Smooth Muscle Cells
[0062] Immunof...
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