Function and application of Vinexin[beta] in treatment of atherosclerosis
A technology of atherosclerosis and β gene, which is applied in the field of gene function and application, can solve the unexplained problems of biological function and possible mechanism of action, and achieve the effect of promoting atherosclerosis
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Embodiment 1
[0041] Example 1 Mouse Atherosclerosis Model (AS) Obtained
[0042] 1. Grouping of experimental animals: 8 weeks old, weighing 19-25g, male, ApoE - / - Mice and Vinexin beta - / - ApoE - / - Mice were fed high-fat diet (Western Diets, HFD) and low-fat diet (Normal chow, NC) respectively, ApoE - / - HFD group, ApoE - / - NC group, Vinexin β - / - ApoE - / - HFD group, Vinexinβ - / - ApoE - / - The NC group consisted of 4 groups with 20 rats in each group.
[0043] 2. Atherosclerosis model induced by high-fat diet:
[0044] Using ApoE - / - Mice and Vinexin beta - / - ApoE - / - In mice, AS models were established, and phenotype correlation analysis was performed to clarify the role of Vinexinβ gene in atherosclerotic diseases. From the age of 8 weeks, the mice in the HFD group were sacrificed after 28 weeks of high-fat diet, and the samples were collected in the NC group after 28 weeks of low-fat diet.
Embodiment 2
[0045] Example 2 Determination of plaque area in AS model mice
[0046] 1. Final mouse tissue harvesting
[0047] 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
[0062] Example 3 Determination of Plaque Stability in AS Model Mice
[0063] 1. Area Size Determination of the Center of Aortic Sinus Necrosis
[0064] Hematoxylin and eosin staining (HE staining) of the paraffin white slices of the aortic sinus was performed in the same manner as in Example 2.4, and the tissues containing cholesterol crystals and fiber structures without nuclei were selected and photographed under a microscope.
[0065] Determination of the area of the necrosis center: use Image-Pro Plus 6.0 image analysis software to circle the area of the necrosis center.
[0066] 2. Determination of collagen content in aortic sinus:
[0067] Sirius red (PSR) staining, the main steps are: take aortic sinus paraffin white slices and 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 1min→mass fraction 0.2% pho...
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