Application of reagent for knocking down or inhibiting CLEC4D in preparation of medicine for myocardial ischemia-reperfusion injury

A technology for reperfusion injury and myocardial ischemia, applied in the field of biomedicine, can solve the problems that CLEC4D protein has no related research and reports

Active Publication Date: 2022-05-06
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there are no related studies and reports on the role and mechanism of CLEC4D protein in myocardial ischemia-reperfusion injury

Method used

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  • Application of reagent for knocking down or inhibiting CLEC4D in preparation of medicine for myocardial ischemia-reperfusion injury
  • Application of reagent for knocking down or inhibiting CLEC4D in preparation of medicine for myocardial ischemia-reperfusion injury
  • Application of reagent for knocking down or inhibiting CLEC4D in preparation of medicine for myocardial ischemia-reperfusion injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] 1. Construction of AAV9-shCLEC4D adeno-associated virus vector

[0039] Utilize restriction endonucleases to digest the interference vector pHBAAV-U6-MCS-CMV-EGFP and the shRNA A fragment (the positive strand nucleotide sequence is shown in SEQ ID NO.1: 5'-AATTCGCTTCCAGTCTAACTGTTACTTCTCGAGAAGTAACAGTTAGACTGGAAGTTTTTTG-3'; reverse strand Nucleotide sequence such as SEQ ID NO.2: 5'-GATCCAAAAAACTTCCAGTCTAACTGTTACTTCTCGAGAAGTAACAGTTAGACTGGAAGCG-3') connection, followed by Escherichia coli transformation, after transformation, use conditional medium (non-resistant LB medium) to screen the target strain and send it to Qingke The company sequenced and used the nucleic acid extraction kit for plasmid extraction to obtain the recombinant adenovirus AAV9-shCLEC4D and the interference vector pHBAAV-U6-MCS-CMV-EGFP. figure 1 As shown, the sequencing results are as follows figure 2 shown. according to figure 2 It can be seen from the sequencing structure that the sequencing resu...

Embodiment 2

[0043] 1. Mouse grouping and model establishment

[0044] 40 experimental mice purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd. were equally divided into myocardial ischemia-reperfusion injury (IRI) group (experimental group) and sham operation (Sham) group (control group);

[0045] Experimental group: each mouse was anesthetized by intraperitoneal injection of 4wt% chloral hydrate at 10 μl / g, and the neck skin, muscle and tissue covering the trachea were separated under a stereoscope. After the trachea was exposed, two tracheal cartilage rings under the glottis Cut a small hole in between, insert the endotracheal tube, and fix it. Check the movement of the thorax to make sure both lungs are well ventilated. Under the stereoscope, use small scissors to make a 1.2cm long incision at the fourth and fifth intercostal spaces on the left edge of the mouse sternum, use micro forceps to bluntly separate the chest wall muscles and intercostal muscles, exp...

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Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of a reagent for knocking down or inhibiting CLEC4D in preparation of a medicine for myocardial ischemia-reperfusion injury. By knocking down the CLEC4D, the CLEC4D protein expression can be inhibited, the cardiac function after myocardial ischemia reperfusion is improved, the cardiac fibrosis after myocardial ischemia reperfusion is reduced, and the myocardial cell hypertrophy after myocardial ischemia reperfusion is improved.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of reagents for knocking down or inhibiting CLEC4D in the preparation of drugs for myocardial ischemia-reperfusion injury. Background technique [0002] Myocardial ischemia-reperfusion injury refers to the pathological process in which the ischemic myocardium recovers to normal perfusion when the coronary artery is partially or completely acutely obstructed and recanalized within a certain period of time, but the tissue damage is progressively aggravated. A series of damaging changes in myocardial ultrastructure, energy metabolism, cardiac function, and electrophysiology caused by ischemia are more prominent after recanalization, and severe arrhythmias may even occur, leading to sudden death. Myocardial ischemia-reperfusion injury can occur in open-heart surgery, coronary artery bypass grafting, transluminal coronary angioplasty, post-thrombolysis, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K31/713A61P9/10A61P9/04
CPCA61K45/00A61K31/713A61P9/10A61P9/04
Inventor 肖俊杰周秋莲刘利孟丹妮
Owner SHANGHAI UNIV
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