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Application of a kind of adenine nucleotide transporter 1

A technology of adenine nucleotide and transporter, which is applied in the application field of adenine nucleotide transporter 1 in the field of cardiac hypertrophy treatment to achieve the effect of inhibiting cardiac hypertrophy

Active Publication Date: 2020-08-04
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the role and mechanism of thiol nitrosylation modification of ANT1 in cardiovascular diseases have not yet been studied or reported.

Method used

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  • Application of a kind of adenine nucleotide transporter 1
  • Application of a kind of adenine nucleotide transporter 1
  • Application of a kind of adenine nucleotide transporter 1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] In order to explore the role of thiol-nitrosylation modification of ANT1 protein in cardiac hypertrophy, we cultured primary cardiomyocytes of neonatal rats, stimulated with Ang II (100 nM) for 48 h to induce hypertrophy of cardiomyocytes, compared with normal saline group, Ang II II (100 nM) stimulation for 48 h can significantly increase the thiol-nitrosylation level of ANT1 ( figure 1). Next, we performed sham surgery (sham) and coarctation of the aorta (TAC) on wild-type C57BL / 6 mice, and extracted sulfhydryl-nitrosylated proteins from myocardial tissue 4 weeks after surgery. The level of thiol-nitrosylation modification of ANT1 protein in mouse myocardial tissue was detected by Biotin-switch. It was found that compared with the control group, the level of thiol-nitrosylation modification of ANT1 protein in the myocardial tissue of mice in the TAC operation group was significantly increased ( figure 2 ). The above experimental results suggest that in the cells a...

Embodiment 2

[0086] In order to further clarify the role of thiol-nitrosylation modification of ANT1 protein in cardiac hypertrophy, we combined liquid chromatography tandem mass spectrometry (LC-MS / MS) technology to screen the thiol-nitrosylation modification of ANT1 in SHR and control mice WKY It was found that in SHR heart tissue, the 57th, 129th and 160th cysteine ​​sites (Cys57, Cys129, Cys160) of ANT1 can undergo sulfhydryl nitrosylation modification ( Figure 3A-Figure 3C ). The 57th, 129th and 160th cysteine ​​sites of ANT1 were mutated to alanine (C57A, C129A, C160A) and then transfected into 293T cells, and it was further concluded that the Cys160 site of ANT1 was modified by sulfhydryl nitrosylation ( Figure 4 ). Construct ANT1 wild-type adenovirus (Ad WT), adenovirus with 160th cysteine ​​mutation (AdC160A) and control group adenovirus (Ad GFP) to infect primary cardiomyocytes, give Ang II stimulation, and detect cardiac hypertrophy The expression of related genes ANP and B...

Embodiment 3

[0088] In order to further verify the role of thiol-nitrosylation modification of ANT1 protein in cardiac hypertrophy, we administered 4-week-old C57BL / 6 mice with tail vein injection of heart-specific overexpressed ANT1 wild-type (aav9-ANT1 WT) and ANT1 The adeno-associated virus with cysteine ​​160 site mutation (aav9-ANT1 C160A) underwent sham and TAC 4 weeks later, and 4 weeks after the operation, the cardiac function and heart-related indicators of the mice were detected by echocardiography, and it was found that after TAC The level of cardiac hypertrophy was significantly increased (IVS-interventricular septal thickness, IVPW-left ventricular posterior wall thickness), and the level of cardiac hypertrophy was significantly reduced after TAC operation with mutation of ANT1 cysteine ​​160 ( Figure 7 ). Heart tissue was collected to measure heart weight, left ventricle weight, mouse tibia length and other indicators. The results showed that after TAC operation with ANT1 cy...

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Abstract

The invention discloses application of adenine nucleotide translocator 1 (ANT1), and relates to the field of the treatment of cardiovascular diseases. Through down-regulating the level of the sulfydryl nitrosylation modification of the ANT1 in a myocardial cell, the occurrence of myocardial hypertrophy is effectively inhibited. A method comprises the following steps of transfecting a myocardial cell by utilizing an adenovirus with the modification site mutation of cysteine, and regulating and controlling the expression of the sulfydryl nitrosylation modification of ANT1 protein in a cell, andalso comprises the following steps of specifically transfecting the heat tissue of a mouse by utilizing an adeno-associated virus with the modification site mutation of the cysteine, and regulating and controlling the expression of the sulfydryl nitrosylation modification of ANT1 protein in the heart tissue. Therefore, the sulfydryl nitrosylation modification of the ANT1 protein can be used as a new important target for clinically treating the myocardial hypertrophy, and has potential clinical application value in the prevention and treatment of the myocardial hypertrophy.

Description

technical field [0001] The invention belongs to the field of cardiovascular disease treatment, and in particular relates to the application of an adenine nucleotide transporter 1 in the field of myocardial hypertrophy treatment. Background technique [0002] With the rapid development of today's society, cardiovascular disease, as the first of the "three major killers" of human health, is gradually reducing the quality of human life and seriously threatening human life and health. Among many cardiovascular diseases, the occurrence and development of cardiac hypertrophy have received widespread attention. At present, it is believed that myocardial hypertrophy is an important "transition" process in the process of heart failure, and it is a compensatory response of the myocardium to mechanical stress. Myocardial hypertrophy is the adaptive change of the myocardium to cardiac insufficiency. The initial compensation has a beneficial effect, and the later decompensation has adve...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68A61K35/761A61P9/04A61P9/00
CPCA61K35/761G01N33/68G01N2800/32
Inventor 季勇唐欣奈拉刘洁琼韩艺谢利平
Owner NANJING MEDICAL UNIV
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