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Functions and applications of nucleotide synthetase CAD in treatment of atherosclerosis

A technology of atherosclerosis and RNA interference, applied in the direction of medical preparations containing active ingredients, gene therapy, drug combinations, etc., can solve the problems of unexplained biological functions and possible mechanisms of action, and achieve the goal of promoting atherosclerosis hardening effect

Active Publication Date: 2015-01-07
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

Since NOD2 regulates the pro-inflammatory activity of macrophages, which are present in early atherosclerotic lesions, we believe that CAD may play an important role in the characteristic pathophysiology of atherogenesis, but Its biological function and possible mechanism of action in the characteristic pathophysiological process of atherosclerosis have not yet been elucidated

Method used

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  • Functions and applications of nucleotide synthetase CAD in treatment of atherosclerosis
  • Functions and applications of nucleotide synthetase CAD in treatment of atherosclerosis
  • Functions and applications of nucleotide synthetase CAD in treatment of atherosclerosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Mouse Atherosclerosis Model (AS) Obtained

[0037] 1. Grouping of experimental animals: 8 weeks old, weighing 19-25g, male, ApoE - / - Mice and CAD - / - ApoE - / - Mice were fed high-fat diet (Western Diets, HFD) and low-fat diet (Normal chow, NC) respectively, ApoE - / - HFD group, ApoE - / - NC group, CAD - / - ApoE - / - HFD group, CAD - / - ApoE - / - The NC group consisted of 4 groups with 20 rats in each group.

[0038] 2. Atherosclerosis model induced by high-fat diet:

[0039] Using ApoE - / - Mice and CAD - / - ApoE - / - In mice, AS models were established, and phenotype correlation analysis was performed to clarify the role of CAD genes 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

[0040] Example 2 Determination of plaque area in AS model mice

[0041] 1. Final mouse tissue harvesting

[0042] 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. Area Size Determination of the Center of Aortic Sinus Necrosis

[0059] For hematoxylin and eosin staining (HE staining), the method is the same as in Example 2.4, and a tissue containing cholesterol crystals and no nucleus fiber structure is selected to take pictures under a microscope.

[0060] 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.

[0061] 2. Determination of collagen content in aortic sinus:

[0062] 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% phosphomolybdic acid for 2min→0.1% Sirius red picric acid solution ...

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Abstract

The invention discloses functions and applications of nucleotide synthetase CAD in treatment of atherosclerosis, belonging to the fields of functions and applications of genes. ApoE- / - mice and CAD- / -ApoE- / - mice are taken as experimental subjects, an atherosclerosis model is obtained through high fat diet induction, the result proves that compared with the ApoE- / - mice, the plaque area of the aorta is obviously reduced due to the CAD gene defects, the stability of sinus plaques of the aorta is enhanced, and the inflammatory response is obviously alleviated. The functions of the CAD in atherosclerosis mainly refer to the effects of promoting aorta plaque formation, particularly the effect of promoting the atherosclerosis. Aiming at the functions of the CAD, the CAD can serve as a drug target to be used for screening medicines for preventing, relieving and / or treating the atherosclerosis, and inhibitors of the CAD can be used for preparing medicines for preventing, relieving and / or treating the atherosclerosis.

Description

[0001] technical field [0002] The invention belongs to the field of gene function and application, and specifically relates to a nucleotide synthetase CAD: carbamoyl-phosphate synthetase II, aspartate aminotransferase and dihydroorotase (carbamoyl-phosphate synthetase II, aspartate The function and application of transcarbamylase, and dihydroorotase (CAD) in the treatment of atherosclerosis, specifically the application of CAD in the preparation of drugs for preventing, alleviating and / or treating atherosclerosis. Background technique [0003] Cardiovascular and cerebrovascular diseases are the main cause of death in many developed countries, and the morbidity and mortality in my country are also increasing year by year. The basis of cardiovascular and cerebrovascular diseases is atherosclerosis (Atheosclersis, AS). Atherosclerosis can thicken and harden the arterial wall and narrow the lumen, leading to many cardiovascular and cerebrovascular events. Acute stenosis and o...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61K48/00A61K39/395A61P9/10
Inventor 李红良赵辉王丕晓程文林
Owner 武汉惠康基因科技有限公司
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