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Function and application of ubiquitin-specific protease 18 (USP18) on treatment of cardiac hypertrophy

A kind of myocardial hypertrophy and protease technology, which is applied to the function and application of genes to achieve the effect of anti-cardiac fibrosis myocardial hypertrophy, inhibition of myocardial hypertrophy and protection of cardiac function

Active Publication Date: 2015-12-30
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, interferon can induce apoptotic response. Studies have suggested that USP18 gene silenced cells can be stimulated by interferon to enhance the pro-apoptotic effect of IFN, but the pro-apoptotic pathways are different in different cell lines.

Method used

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  • Function and application of ubiquitin-specific protease 18 (USP18) on treatment of cardiac hypertrophy
  • Function and application of ubiquitin-specific protease 18 (USP18) on treatment of cardiac hypertrophy
  • Function and application of ubiquitin-specific protease 18 (USP18) on treatment of cardiac hypertrophy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] The expression of embodiment 1 USP18 in the heart of normal person and patient with cardiomyopathy

[0080] 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 USP18 protein and cardiomyocyte hypertrophy markers ANP (Millipore, AB2232), β-MHC (santacruz, sc53090), and the expression of USP18 (santacruz, sc98431), GAPDH (CellSignalingTechnology, 2128) as an internal reference. Test results such as figure 1 As shown, the expression of cardiomyocyte hypertrophy markers ANP and β-MHC in the hearts of patients with dilated cardiomyopathy was significantly up-regulated, and the expression of USP18 was also significantly up-regulated ( figure 1 ).

Embodiment 2

[0081] Example 2 Expression of USP18 in the heart of wild-type mouse sham operation group and cardiac hypertrophy model group

[0082] 1. Aortic arch constriction (AB) was used to establish a mouse model of myocardial hypertrophy. The model operation process:

[0083] 1.1 Preoperative preparation

[0084] (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).

[0085] (2) Preparation of the operation area: the skin of the left chest, left chest and armpit of the left forelim...

Embodiment 3

[0096] Example 3 Effect of USP18 interference (AdshUSP18) and overexpression (AdUSP18) adenovirus on AngII-stimulated primary cardiomyocyte hypertrophy

[0097] 1. Primary neonatal SD rat cardiomyocyte culture

[0098] (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.

[0099] (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.

[0100] (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 seru...

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Abstract

The invention discloses a function and application of ubiquitin-specific protease 18 (USP18) on treatment of cardiac hypertrophy, and belongs to the field of functions and application of genes. A mutual relation between USP18 expression and the cardiac hypertrophy is determined, and a research result shows that in a model suffering from the cardiac hypertrophy, USP18 expression is obviously lower than that of a normal group; and cardiac hypertrophy and fibrosis are promoted and a cardiac function is worsened by inhibiting USP18 expression, and cardiac hypertrophy and fibrosis are obviously inhibited and the cardiac function is protected by promoting USP18 overexpression. Therefore, the USP18 can serve as a target gene, is used for screening medicines for protecting the cardiac function, resisting cardiac fibrosis and / or preventing and relieving and / or treating cardiac hypertrophy, and is used for preparing medicines for protecting the cardiac function, resisting cardiac hypertrophy and / or preventing and relieving and / or treating cardiac hypertrophy, and a new effective path is provided for treatment of cardiac hypertrophy 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 protease 18 (USP18) in the treatment of cardiac hypertrophy. Background technique [0002] Myocardial hypertrophy is the compensatory response of the myocardium to long-term biomechanical pressure or increased volume load, which is commonly seen in cardiovascular diseases such as hypertension and aortic stenosis. Increased and other characteristics [1-3]. Hypertension and senile degenerative aortic valve disease are on the rise year by year in our country. Cardiac hypertrophy and the incidence of hypertension and heart disease caused by hypertension and other diseases also increase thereupon. Although myocardial hypertrophy can initially increase myocardial cells and strengthen myocardial contractility, which is a compensatory mechanism to maintain normal cardiac output, long-term continuous pressure or ...

Claims

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Application Information

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IPC IPC(8): A61K38/46A61K45/00A61P9/00G01N33/68
Inventor 李红良何奔应小盈
Owner WUHAN UNIV
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