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 pa

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

Example Embodiment

[0036] Example 1 Obtaining a mouse atherosclerosis model (AS)

[0037] 1. Experimental animal grouping: select 8 weeks old, weight 19-25g, male, ApoE - / - Mice and CAD - / - ApoE - / - Mice were fed with high-fat diet (Western Diets, HFD) and low-fat diet (Normal chow, NC), ApoE - / - HFD group, ApoE - / - NC group, CAD - / - ApoE - / - HFD group, CAD - / - ApoE - / - There are 4 groups in the NC group, with 20 animals in each group.

[0038] 2. The process of atherosclerosis model induced by high-fat feed:

[0039] Adopt ApoE - / - Mice and CAD - / - ApoE - / - Mice, establish an AS model, perform phenotypic correlation analysis, and clarify the role of CAD gene on atherosclerotic disease. Starting from 8 weeks of age, mice in the HFD group were sacrificed after being fed a full course of high-fat diet for 28 weeks and samples were collected. The NC group was sacrificed after being fed a full course of low-fat diet for 28 weeks and samples were collected.

Example Embodiment

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

[0041] 1. Mouse terminal tissue collection

[0042] The mice were fed high-fat or low-fat feed until 28 weeks, weighed, and anesthetized the mice with 3% sodium pentobarbital, 90 mg / kg, fixed with a needle on the sample plate, and moistened the chest and abdomen skin of the mice with gauze. Ophthalmic scissors cut open the chest cavity, expose the heart, cut open the right atrial appendage, pierce the needle of the infusion set into the left ventricle, and slowly inject 10-15mL PBS buffer with a 50mL syringe. When the right atrial appendage is clear, replace it with 4% poly Continue to inject 10-15 mL of formaldehyde. After the perfusion, the organs in the thoracic and abdominal cavity were removed, leaving only the heart. Place the mouse under a microscope, separate the fascia and adipose tissue around the aortic arch, cut off the head and arm trunk, put it into a 5mL EP tube containing 4% paraformaldehyde, cut the h...

Example Embodiment

[0057] Example 3 Determination of plaque stability in AS model mice

[0058] 1. Determination of the area of ​​the center of aortic sinus necrosis

[0059] Hematoxylin and eosin staining (HE staining), the method is the same as that in Example 2.4, and the tissues containing cholesterol crystals and no nuclear fiber structure are selected and photographed under a microscope.

[0060] Determination of the area of ​​the necrotic center: Use Image-Pro Plus 6.0 image analysis software to circle the area of ​​the necrotic 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 min → xylene for 2 min, 3 times → 100% alcohol for 1 min → 95% alcohol for 1 min → 70% alcohol for 1 min → running water for 10 min → Double distilled water for 1 min→mass fraction 0.2% phosphomolybdic acid for 2 min→0.1% Sirius scarlet picric acid solution is dripped on the tissue, st...

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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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