Application of CFTR gene to preparation of thrombotic disease early warning or diagnosing medicine

A thrombotic disease, 1. CFTR technology, applied in the application field of CFTR gene in the preparation of early warning or diagnosing thrombotic disease drugs, can solve the problems of lack of specific diagnosis, affecting the quality of life and economic burden of patients, lack of specificity, etc.

Inactive Publication Date: 2019-05-10
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Acute Coronary Syndrome (ACS) is one of the acute and critical diseases of coronary heart disease. Its morbidity and mortality are increasing year by year, seriously affecting the quality of life and economic burden of pat

Method used

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  • Application of CFTR gene to preparation of thrombotic disease early warning or diagnosing medicine
  • Application of CFTR gene to preparation of thrombotic disease early warning or diagnosing medicine
  • Application of CFTR gene to preparation of thrombotic disease early warning or diagnosing medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Changes in CFTR protein levels of platelets in peripheral blood of normal people and patients with coronary heart disease

[0017] (1) Materials and methods

[0018] 1. Collection and grouping of peripheral blood of clinical patients

[0019] Peripheral blood from the population was collected from Guangdong Provincial People's Hospital after approval by the Ethics Committee of Guangdong Provincial People's Hospital. According to the clinical diagnosis index, it is divided into Non-CAD group (non-coronary artery disease group), SA group (stable angina pectoris group), ACS group (acute coronary syndrome group)

[0020] 2. Human Platelet Preparation

[0021] After disinfecting the elbow skin of the volunteers, draw venous blood, put the blood into an anticoagulant tube, anticoagulate the blood and anticoagulant with sodium citrate at a ratio of 9:1; centrifuge at 300g for 3 minutes at room temperature, and gently draw the upper Put platelet rich plasma (Platel...

Embodiment 2

[0028] Example 2 Changes of CFTR protein levels in peripheral blood mononuclear cells of normal people and patients with coronary heart disease

[0029] (1) Materials and methods

[0030] 1. Decreased expression of CFTR on monocytes of patients with ACS

[0031] (1) Preparation of peripheral blood mononuclear cells: The liquid (1.5 ml) in the anticoagulated blood sample was gently mixed with D-Hank’s buffer, and the total volume was 3 ml. In a 15ml centrifuge tube, carefully pour the diluted sample onto the surface of 3ml of Ficoll solution. To obtain peripheral blood mononuclear cells, samples were centrifuged at 800g×20min at room temperature. After centrifugation, a mononuclear cell layer should appear between the Ficoll solution and the plasma. Collect monocytes (1mL) and wash with 8ml of D-Hank's buffer before centrifugation at 300g×5min, and repeat twice. The monocytes were then resuspended in serum-free RPMI 1640 medium and seeded at 35 mm 2 Petri dish. Cells wer...

Embodiment 3

[0035] Example 3 Ferric chloride-induced mesenteric arteriolar thrombosis model

[0036] (1) Materials and methods

[0037] Experimental principle: The model of mouse mesenteric arteriole thrombosis induced by FeCl3 is to label mouse platelets with fluorescent dye (Calcein-AM, calceinacctoxymethyester) in vitro, inject tail vein back into experimental or control mice, and treat with FeCl3 The mesenteric arterioles injure the vascular endothelium, induce the formation of thrombus at the damaged site, and observe the formation of arterial thrombus in real time under the fluorescence microscope.

[0038] 1. Mice were anesthetized with 10% sodium pentobarbital

[0039] 2. Fix the mouse on the operating table, separate the abdominal aorta of the mouse, and use a 2ml syringe to take blood from the abdominal aorta. Add 3.8% sodium citrate anticoagulant to the syringe in advance, and the ratio of blood to anticoagulant The ratio is 9:1. After taking the blood, transfer the blood to ...

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Abstract

The invention discloses application of a CFTR gene to preparation of thrombotic disease early warning or diagnosing medicine. The CFTR gene codes a chloride ion channel with cAMP dependency, and it isfound that the expression of the CFTR gene on the peripheral blood platelets and mononuclear cells of a patient with a coronary heart disease is reduced remarkably; statistics difference is shown among patient groups with stable angina pectoris and acute coronary syndromes. On an animal cell model, the specificity restraining CFTR gene is found to remarkably promote thrombosis, and has the effects of promoting platelet aggregation and up regulating a P2Y12 receptor-mediated platelet aggregation signal channel. The new medical application of the CFTR gene to thrombotic disease early warning and diagnosing is mined, the new application field of gene treatment is expanded, and the new clinical diagnosis marker is provided for early warning and diagnosing of a critical thrombotic disease.

Description

technical field [0001] The invention relates to the application of CFTR gene in early warning and diagnosis of thrombotic diseases, and the use of CFTR gene to screen possible drug use for treating thrombotic diseases. Background technique [0002] Cystic fibrosis transmembrane conductance regulator (CFTR) is a type of ATP-gated, cAMP-dependently activated chloride ion channel, and its basic physiological function is to regulate chloride ion secretion and transmembrane material transport. Mutations in the CFTR gene lead to the development of cystic fibrosis (CF), a rare but fatal autosomal recessive disorder that primarily affects the function of the respiratory, intestinal, and reproductive glandular epithelium. In the circulatory system, vascular smooth muscle (including aorta, pulmonary artery, mesenteric artery, cerebral basilar artery, etc.), vascular endothelium, and platelets have CFTR chloride channel mRNA and protein expression. It has been reported that CFTR gene ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883
Inventor 王冠蕾杨涵滟
Owner SUN YAT SEN UNIV
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