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
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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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