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Application of leukocyte immunoglobulin like receptor B4 (LILRB4) to preparation of medicine for preventing, alleviating and/or treating cardiac hypertrophy

A technology of immunoglobulin and myocardial hypertrophy, which is applied in the field of gene function and application, can solve problems such as LILRB4 that has not been seen, and achieve the effect of inhibiting myocardial hypertrophy and improving heart function

Active Publication Date: 2018-06-22
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there is no report on the role of LILRB4 in pathological cardiac hypertrophy

Method used

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  • Application of leukocyte immunoglobulin like receptor B4 (LILRB4) to preparation of medicine for preventing, alleviating and/or treating cardiac hypertrophy
  • Application of leukocyte immunoglobulin like receptor B4 (LILRB4) to preparation of medicine for preventing, alleviating and/or treating cardiac hypertrophy
  • Application of leukocyte immunoglobulin like receptor B4 (LILRB4) to preparation of medicine for preventing, alleviating and/or treating cardiac hypertrophy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] [Example 1] Construction of heart-specific LILRB4 knockout mice:

[0046] Using CRISPR-Cas9 technology to construct heart-specific LILRB4 gene knockout mice (for the construction strategy, see figure 1 ). First, a CRISPR targeting site was designed in the front non-coding region of exon 1 and intron 2 of the mouse LILRB4 gene through the online CRISPR design tool (http: / / crispr.mit.edu), and the target sequences were respectively for:

[0047] LILRB4sgRNA1:ggTAGGATGTCAGAGTTGCAGCGTGG,

[0048] LILRB4sgRNA2: ggATTCCTCAAAATGGTGATGGGGGG.

[0049] In addition, a donor vector (Donor Vector) for homology repair was designed, which included homology arms on both sides, exons 1 and 2 in the middle, and two loxp sequences in the same direction.

[0050] (1) Construction of targeting vector: The two primers corresponding to sgRNA1 and sgRNA2 were fused into double-stranded DNA, and then ligated into pUC57-sgRNA vector treated with restriction endonuclease BsaI with T4 DNA liga...

Embodiment 2

[0066] [Example 2] Establishment of mouse cardiac hypertrophy model

[0067] 1. Grouping of experimental animals: male background C57BL / 6 cardiac-specific Cre mice (α-MHC-Cre (WT), cardiac-specific LILRB4 gene knockout mice (LILRB4-KO), established myocardial muscle by aortic constriction Hypertrophy model. Randomly divided into 4 groups, grouped as follows: C57BL / 6 background wild-type mice sham operation group (WT Sham) and AB operation group (WT AB), LILRB4 gene knockout mouse sham operation group (LILRB4-KO Sham ) and AB group (LILRB4-KO AB).

[0068] 2. The myocardial hypertrophy model adopts aortic arch coarctation (AB) surgery, and the operation process of the model is as follows:

[0069] 2.1 Preoperative preparation

[0070] (1) Anesthesia: weigh the mice first, 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 t...

Embodiment 3

[0077] [Example 3] Myocardial hypertrophy and fibrosis evaluation of mouse cardiac hypertrophy model

[0078] 1. Take materials

[0079] (1) Preliminary work: prepare in advance a urine cup filled with 20 mL of 10% formaldehyde by volume, and label it (mouse number, group, type of operation and date of collection). Place a petri dish filled with 10% KCl solution in mass fraction at the sampling site. Turn on the analytical balance and set it to zero for later use. Mice were then weighed and sacrificed.

[0080] (2) Material collection: Ophthalmic curved tweezers clamped the vascular pedicle below the atrial appendage, cut off the heart, and quickly placed it in 10% KCl solution by mass fraction. After the heart stops beating in the diastolic phase, place it on sterilized gauze, gently squeeze the fluid in the heart cavity, dip the surface fluid dry, weigh and record, put the heart into the corresponding urine cup, fix it for 48 hours and use it for pathological testing.

...

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Abstract

The invention discloses application of leukocyte immunoglobulin like receptor B4 (LILRB4) to preparation of medicine for preventing, alleviating and / or treating cardiac hypertrophy. Cardiovascular-specific Cre mice and cardiovascular-specific LILRB4 gene knockout mice are selected, a surgery of coarctation of aorta is carried out by an operation group, determination of cardiac hypertrophy, fibrosis and cardiac functions on various groups of mice can be carried out, and a result proves that the cardiac hypertrophy, the fibrosis and the cardiac functions are obviously worsened because of LILRB4defects caused by knockout LILRB4 genes. Through recombination of adenovirus, AdshLILRB4 and AdLILRB4 infected SD primary myocardial cells of suckling mice are built, and a cardiomyocyte hypertrophy model is built through Ang II stimulation; due to LILRB4 over-expression viruses, the cardiomyocyte hypertrophy is obviously inhibited, the surface area of cardiomyocyte is reduced, and a result of LILRB4 interference virus is opposite. LILRB4 has effects of inhibiting the cardiac hypertrophy and the fibrosis and improving the cardiac functions.

Description

technical field [0001] The invention belongs to the field of gene function and application, in particular to the function and application of leukocyte immunoglobulin like receptor subfamily B member 4 (leukocyte immunoglobulin like receptor B4, LILRB4) in the treatment of cardiac hypertrophy, specifically in the preparation of anti- The application in the medicine of myocardial fibrosis and / or prevention, alleviation and / or treatment of myocardial hypertrophy. Background technique [0002] Myocardial hypertrophy is an adaptive compensatory response of myocardial cells to the stimulation of neurohumoral factors and changes in mechanical stress load. It is a complex dynamic process accompanied by many factors involved in regulation. Cardiomyopathy, cardiomyopathy and other common pathological processes that need to be experienced in the development of most cardiovascular diseases[1]. Myocardial hypertrophy is an adaptive compensatory response of the heart to various cardiovas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K45/00A61K48/00A61P9/00
CPCA61K38/1774A61K45/00A61K48/005
Inventor 李红良
Owner WUHAN UNIV
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