Method for constructing rat myocardial delayed ischemia reperfusion injury model

A technology of ischemia-reperfusion and construction method, which is applied in the field of establishment of rat myocardial delayed ischemia-reperfusion injury model, can solve problems such as difficulty in maintaining consistent needle depth and width, ligation of LAD blood vessels, failure of modeling, etc., to achieve Visible ligation strength, high repeatability, and improved success rate

Inactive Publication Date: 2019-04-19
THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 1. There is no obvious anatomical structure reference for the position of the needle in and out, and it is difficult to keep the depth and width of the needle consistent, and it is often impossible to successfully ligate the LAD blood vessel at one time, resulting in the failure of modeling;
[0008] 2. When reperfusion needs to open the knot again, it will be pulled to the heart many times, and the risk of vascular injury, modeling failure and operation time will increase;
[0009] 3. The hard plastic tube (polyethylene tube) has a smooth and hard surface, so it is difficult to maintain consistent ligation tightness, which increases the risk of vascular injury;
[0010] 4. Previous methodologies did not explore and report on the feasible delayed reperfusion time

Method used

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  • Method for constructing rat myocardial delayed ischemia reperfusion injury model
  • Method for constructing rat myocardial delayed ischemia reperfusion injury model
  • Method for constructing rat myocardial delayed ischemia reperfusion injury model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] A kind of embodiment of the construction method of rat myocardial delayed ischemia-reperfusion injury model of the present invention, comprises the steps:

[0048] (1) SPF grade SD mice were fasted overnight after a 3-day adaptation period, and weighed before operation. Anesthetized by ether inhalation combined with 10% chloral hydrate intraperitoneal administration at a dose of 0.35ml / 100g until the contraction reflex of the lower limbs disappeared. A heating pad was placed under the rat and adjusted to maintain the temperature at 37°C. Shave the hair on the neck and chest area, disinfect with 75% alcohol, and spread a towel. Use curved forceps to properly pull out the tongue to expose the opening of the trachea, and light illuminates the neck to guide the 18G indwelling needle to perform oral tracheal intubation. The outer cannula is connected to a ventilator. Ventilator parameters: positive pressure ventilation, tidal volume 2.5-3ml / 100g, frequency 70-80 / min. Limb...

Embodiment 2

[0053] Previous reports found that after the LAD was continuously occluded for more than 24 hours, even if the blood vessels were opened, the reperfusion injury was not obvious. In order to explore the infarction time point of delayed reperfusion in myocardial infarction with better reproducibility, the inventors performed reperfusion after different ischemia times, which were 1h, 3h, 4h, 6h, 12h and 24h. Under the conditions of this experiment, the delayed ischemia-reperfusion model was explored. Under the condition of ensuring effective reperfusion, the inventor gradually shortened the ischemia time from 24 hours of continuous myocardial infarction. In the 24-hour ischemia group, the surface color of the myocardium was found to be fixed after thoracotomy by the original route, and the adhesions in the pericardium and thoracic cavity were severe. There was no obvious change in the color of the myocardium during reperfusion, no change in the electrocardiogram, and no reperfusio...

Embodiment 3

[0057] Using the model construction method in Example 1, in the exploration of delaying ischemic time, the inventor tried to give rats unfractionated heparin (50UI / 100g, i.p.) to prolong the ischemic time before operation, and found that it would significantly increase incision bleeding and pleural volume. Asphyxiation due to blood or intratracheal hemorrhage. Moreover, the blurred surgical field is not conducive to observing the color of the myocardium, so it is more feasible to consider reperfusion after 4hrs ischemia. In the delayed I / R model, the TTC / Evens blue double staining method to mark the ischemic infarction area, risk area and normal area has the advantages of economy, strong repeatability, and good contrast; it is the most widely used detection method for evaluating reperfusion injury . Take the 2mm horizontal myocardial cross-section under the ligation point for coloring analysis, the inventors found that the ischemic involvement area in the delayed I4h / R group ...

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Abstract

The invention discloses a method for constructing a rat myocardial delayed ischemia reperfusion injury model, which comprises the following steps: (1) taking a rat for fasting overnight, anaesthetizing, passing through an oral trachea cannula through a 18G indwelling needle, connecting an outer sleeve with a breathing machine, and connecting four limbs with leads of an electrocardiograph; (2) longitudinally cutting skin from the left side of the sternum, lightly tearing the pericardium, inserting a needle at the root of the pulmonary artery cone, and discharging a needle thread at the position2-3mm below the left auricle; (3) placing a self-made rubber PE section on the surface of the heart at the ligation position, knotting on the rubber PE section, and performing continuous ligation ischemia for 1-6 hours in the myocardial infarction group; (4) opening the chest again to expose the ligation point, and observing the color of the anterior wall myocardium 15-20 seconds after the re-perfusion; suturing with 4-0 surgical suture after thoracotomy, continuous auxiliary ventilating and performing ibuprofen analgesia to obtain the rat myocardial delayed ischemia reperfusion injury model.According to the method, the natural rubber PE section is used as a medium for pressing the LAD in the model, so that the method has the advantages of rapid re-irrigation, visible ligation force andthe like; according to the method, a reperfusion injury model with high repeatable rate can be obtained after 4 hrs of ischemia.

Description

technical field [0001] The invention relates to the technical field of animal model construction, in particular to a method for constructing a rat myocardial delayed ischemia-reperfusion injury model. Background technique [0002] With the aging of the population, the incidence of myocardial infarction (MI) is also increasing year by year. Persistent ischemia caused by acute occlusion not only causes permanent damage to the myocardium, impaired ventricular systolic function, but also leads to the occurrence of chronic heart failure. With the continuous development of interventional technology, early reperfusion of damaged myocardium is the main treatment option. As a continuous pumping organ, the heart is particularly sensitive to ischemia and hypoxia. Even after effective reperfusion, cardiomyocytes still have obvious necrosis and apoptosis, which is called ischemia reperfusion injury (IRI). [0003] Under current conditions, the timely reperfusion of acute myocardial inf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61D1/00
CPCA61D1/00
Inventor 崔同涛唐咏文罗碧辉王涛陆东风谭宁
Owner THE FIRST AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV (GUANGZHOU RESPIRATORY CENT)
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