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Function and application of ubiquitin specific protease 4 (USP4) in treating cardiac hypertrophy

A kind of myocardial hypertrophy and specific technology, applied in the field of gene function and application, to achieve the effect of protecting heart function

Active Publication Date: 2016-01-20
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

Current studies have shown that USP4 mainly plays the roles of inhibiting inflammatory response, anti-virus, regulating cell cycle and promoting tumorigenesis, but its role in the heart has not been reported yet. Active implementation of signaling pathways (17,22,23)

Method used

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  • Function and application of ubiquitin specific protease 4 (USP4) in treating cardiac hypertrophy
  • Function and application of ubiquitin specific protease 4 (USP4) in treating cardiac hypertrophy
  • Function and application of ubiquitin specific protease 4 (USP4) in treating cardiac hypertrophy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] The expression of embodiment 1 USP4 in the heart of normal person and patient with cardiomyopathy

[0079] SDS-PAGE-immunoblotting test was performed on proteins extracted from the heart by selecting normal human hearts (individuals donated by non-cardiac causes of death, Donor) and hearts of patients with dilated cardiomyopathy (recipients replaced by patients undergoing heart transplantation, DCM). Western blot), combined with antibodies that specifically recognize USP4 protein and cardiomyocyte hypertrophy markers ANP (Millipore, AB2232), Myh7 (santacruz, sc53090), and the expression of USP4 (CST, 2651), GAPDH (santacruz, sc25778) as an internal reference. Test results such as figure 1 As shown, the expression of cardiomyocyte hypertrophy markers ANP and Myh7 in the hearts of patients with dilated cardiomyopathy was significantly up-regulated, and the expression of USP4 was significantly down-regulated.

Embodiment 2

[0080] Example 2 Expression of USP4 in the heart of wild-type mouse sham operation group and cardiac hypertrophy model group

[0081] 1. Aortic arch constriction (AB) was used to establish a mouse model of myocardial hypertrophy. The model operation process:

[0082] 1.1 Preoperative preparation

[0083] (1) Anesthesia: First weigh the mice, calculate the required amount of anesthetic (3% pentobarbital sodium) according to 90 mg / kg body weight, inject intraperitoneally, and record the injection time point. There is no obvious reaction between tail and toe pinching and the mouse is in good condition. This is the standard for successful anesthesia (generally there is no obvious reaction about 10 minutes after injection, and the mouse has a reaction to pinch toe about 50 minutes after anesthesia, and about 30 minutes after anesthesia is the best operation time).

[0084] (2) Preparation of the operation area: the skin of the left chest, left chest and armpit of the left forelimb...

Embodiment 3

[0095] Example 3 Effect of USP4 interference (AdshUSP4) and overexpression (AdUSP4) adenovirus on AngII-stimulated primary cardiomyocyte hypertrophy

[0096] 1. Primary neonatal SD rat cardiomyocyte culture

[0097] (1) Eight newborn Sprague-Dawley suckling mice were disinfected with 75% alcohol below the neck, and the heart was removed with ophthalmic scissors and micro forceps, and placed in a glass plate filled with 10mL DMEM / F12 solution. Take another one and repeat the above process.

[0098] (2) Wash the heart with DMEM / F12 medium, and cut the heart into 1-2mm 3 fragments. Transfer to a serum bottle with a rotor, suck off DMEM / F12, and add trypsin digestion solution. Rotate at 120r / min, digest for 15min, rest for a few seconds, and discard the supernatant.

[0099] (3) Add trypsin digestion solution, the speed is 120r / min, and digest for 15min. Stand still for a few seconds, aspirate the supernatant, terminate the digestion with DMEM / F12 medium with 20% calf serum, ...

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Abstract

The invention discloses a function and application of ubiquitin specific protease 4 (USP4) in treating cardiac hypertrophy, and belongs to the field of gene functions and application. The mutual relation between the expression of the USP4 and cardiac hypertrophy is determined; research results show that in a cardiac hypertrophy occurrence model, the expression of the USP4 is significantly reduced compared with a normal group; by restraining the expression of the USP4, the cardiac hypertrophy and fibrosis are significantly promoted, and the cardiac function is deteriorated; by promoting the expression of the USP4, cardiac hypertrophy and fibrosis are significantly restrained, and the cardiac function is protected. Accordingly, the USP4 can be used as a target gene for screening and preparing the medicine which protects the cardiac function, resists cardiac fibrosis and / or prevents, relieves and / or treats cardiac hypertrophy, and an effective new way is provided for treating cardiac hypertrophy.

Description

technical field [0001] The invention belongs to the field of gene function and application, and in particular relates to the function and application of ubiquitin specific peptidase 4 (USP4) in the treatment of cardiac hypertrophy. Background technique [0002] Heart failure remains the leading cause of death worldwide, with 5- and 10-year survival rates of only 50% and 10% (1-3). The prevention and treatment of heart failure is a major problem to be solved urgently. Pathological cardiac hypertrophy is a key pathophysiological event in the progression of the heart from compensatory function to heart failure (4). The occurrence and development of pathological myocardial hypertrophy involves a series of changes in extracellular molecules, cell membrane receptors, intracellular signaling pathways, gene expression, cell morphology, gross shape, and cardiac function (5). In mediating the development of pathological myocardial hypertrophy, the most classic pathways include calci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/46A61P9/00G01N33/68
Inventor 李红良何奔赵怡超
Owner 武汉惠康基因科技有限公司
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