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Application of histone methyltransferase EZH2 to preparation of medicinal preparation for preventing or treating aortic diseases

A technology of methyltransferase and pharmaceutical preparation, applied in the field of protein function, can solve the problem that the use of EZH2 has not been reported yet.

Inactive Publication Date: 2018-06-01
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there is no report on the use of EZH2 in the treatment of aortic medial degenerative disease

Method used

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  • Application of histone methyltransferase EZH2 to preparation of medicinal preparation for preventing or treating aortic diseases
  • Application of histone methyltransferase EZH2 to preparation of medicinal preparation for preventing or treating aortic diseases
  • Application of histone methyltransferase EZH2 to preparation of medicinal preparation for preventing or treating aortic diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Collection of samples from patients with aortic dissection and expression level of EZH2

[0039] 16 cases of thoracic aorta tissue were obtained from aortic lesion vessel specimens excised from patients undergoing surgery for acute Stranford type A thoracic aortic dissection in a hospital, and 15 normal controls were obtained from heart transplant patients without history of vascular disease. Echocardiography, chest CT, and angiography confirmed that the aorta was not dilated. After collection, the samples were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator. First, we extracted the total protein in the aortic wall tissue for Western blot detection. The primary antibody was incubated with LC3I / II (CST, #8457) and GAPDH (CST, #5174), and the secondary antibody was incubated with Jackson ImmunoResearch Laboratories, 111-035-003 Incubation. The results showed that autophagy was significantly enhanced in the aortic wall tissue of patients wi...

Embodiment 2

[0045] Example 2 Effect of lentivirus overexpression of EZH2 (lenti-EZH2) on the survival of vascular smooth muscle cells

[0046] First, we established a mouse aortic smooth muscle cell (MOVAS) line overexpressing EZH2. Mouse aortic vascular smooth muscle cells (MOVAS cells) were purchased from ATCC cell bank (CRL-2797TM), using 10% fetal bovine serum (SH30084.03, Hyclone), 1% penicillin-streptomycin double antibody (15140-122 , ThermoFisher Scientific) DMEM high glucose medium (SH30022.01, Hyclone) for cultivation. Cells were infected with lentivirus lenti-EZH2, lenti-GFP, lenti-mCherry-GFP-LC3 for 48 hours, treated with puromycin for 7 days, and cell selection was performed to obtain MOVAS cells successfully infected with lentivirus.

[0047] The total protein of the above cells was extracted, and Western blot was used to detect the protein expression levels of EZH2 and its downstream molecules H3K27me2 and H3K27me3 in the control group and the experimental group. express...

Embodiment 3

[0049] Example 3 Effect and mechanism of targeted activation of EZH2 on autophagy of vascular smooth muscle cells

[0050] The latest research shows that the level of autophagy is an important factor affecting the survival status of vascular smooth muscle cells. Therefore, rapamycin with a final concentration of 150nM was used as an autophagy inducer to stimulate mouse aortic vascular smooth muscle cells, and the cells were treated with dimethyl sulfoxide (DMSO) as a control group, and the total protein was extracted for Western blot experiments to detect EZH2 protein expression levels.

[0051] The results showed that after the vascular smooth muscle cells were stimulated by rapamycin, the expression level of EZH2 protein decreased significantly. At the same time, MOVAS cells were infected and cultured with Lenti-GFP and lenti-EZH2 respectively, and the total protein of cells was extracted for Western blot experiment to detect the expression level of autophagy marker molecul...

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Abstract

The invention discloses application of histone methyltransferase EZH2 to preparation of a medicinal preparation for preventing or treating aortic diseases. The invention confirms that the EZH2 can effectively inhibit autophagy of vascular smooth muscle cells and promote the survival of smooth muscle cells; a novel target spot and strategy are provided for effective treatment of aortic dissection and other aortic diseases with reduction of the vascular smooth muscle cells as the basis of cytology, and a basic is provided for the development of a novel medicament for treating diseases of the type.

Description

technical field [0001] The present invention relates to the field of protein function, and specifically refers to the application of a histone methyltransferase EZH2 in the preparation of pharmaceutical preparations for preventing or treating aortic diseases. Background technique [0002] Aortic medial degenerative disease refers to a series of aortic diseases with aortic medial disease as the pathological basis, including aortic aneurysm, aortic dissection, aortic atherosclerosis, genetic diseases such as Marfan syndrome, etc. general term. Compared with other diseases of the arterial system, aortic disease has a longer subclinical development period and has the clinical characteristics of acute onset. Once symptoms appear, it is very dangerous. Aortic disease is difficult to treat, with high cost, poor treatment effect and high mortality rate. Among them, aortic aneurysm and aortic dissection are the most serious and have the highest mortality rate. The 2010 Global Burde...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/45A61P9/00A61P9/10A61P35/00
CPCA61K38/45C12Y201/01043
Inventor 蒋丁胜魏翔霍博李睿朱学海
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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