The function and application of blood shear stress response protein 1 in the treatment of atherosclerosis
A technology for atherosclerosis and purpose, 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
[0043] Example 1 Obtaining a mouse atherosclerosis model (AS)
[0044] 1. Grouping of experimental animals: 8-week-old, weighing 19-25g, male, ApoE - / - Mice and RECS1 - / - ApoE - / - Mice were fed with high-fat diet (Western Diets, HFD) and low-fat diet (Normal chow, NC), respectively, ApoE - / - HFD group, ApoE - / - NC Group, RECS1 - / - ApoE - / - HFD group, RECS1 - / - ApoE - / - There were 4 groups in the NC group, with 20 animals in each group.
[0045] 2. The atherosclerosis model is induced by high-fat diet. The operation process:
[0046] Using ApoE - / - Mice and RECS1 - / - ApoE - / - In mice, an AS model was established, and phenotype correlation analysis was performed to clarify the role of RECS1 gene on atherosclerotic disease. From 8 weeks old, the mice in the HFD group were sacrificed and the samples were collected after being fed with the high-fat diet for 28 weeks, and the mice in the NC group were sacrificed and the samples were fed with the whole low-fat diet for 2...
Embodiment 2
[0047] Example 2 Determination of plaque area in AS model mice
[0048] 1. Mice terminal tissue harvesting
[0049] Mice were fed with high-fat or low-fat diet until 28 weeks, weighed, anesthetized the mice with 3% sodium pentobarbital, 90 mg / kg, fixed them on the sampling plate with a needle, and wet the skin of the chest and abdomen of the mice with gauze. Cut the chest cavity with ophthalmic scissors, expose the heart, cut open the right atrial appendage, insert the needle of the infusion set into the left ventricle, and slowly inject 10-15 mL of PBS buffer with a 50 mL syringe. Continue to inject 10-15mL of formaldehyde. 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 heart was cut at the beginning of the ascending aorta, the middle of the thoracic aorta, and about 3 mm below the common neck and clavicle ...
Embodiment 3
[0062] Example 3 Determination of plaque stability in AS model mice
[0063] 1. Determination of the area size of the necrotic center of the aortic sinus
[0064] Hematoxylin and eosin staining (HE staining), the method was the same as that of Example 2.4, and the tissue containing cholesterol crystals and no nucleus fibrous structure was selected to take pictures under a microscope.
[0065] Measurement of the area of the necrotic center: The area of the necrotic center was circled using Image-Pro Plus 6.0 image analysis software.
[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℃ for 30min→xylene for 2min, 3 times→100% alcohol for 1min→95% alcohol for 1min→70% alcohol for 1min→rinse with running water for 10min→ Double-distilled water for 1min→0.2% phosphomolybdic acid for 2min→0.1% Sirius scarlet picric acid solution was dropped on the tissue, st...
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