大鼠新型气管插管方法的建立与Epo对心肺复苏后心肌的影响

发布时间：2018-05-16 19:25

本文选题：心脏骤停 + 心肺复苏　；参考：《广州医学院》2009年硕士论文

【摘要】： 背景大鼠是常用的实验研究动物,但由于体型小,口腔狭窄,暴露声门难度大,大鼠气管插管一直是个难题。文献报道的大鼠气管插管方法主要有:气管切开插管法,盲插法,直视下插管法。既往的方法都它的局限性,容易出现插管失败和并发症。因此,有必要寻求一种新的插管方法,实现安全、准确的大鼠气管插管。我们利用气管导管内置入直径为0.9mm的光纤作为光源和导丝,建立一种新型大鼠气管插管方法。复苏后心功能不全主要表现为低血压、休克、致命性的室性心律失常或再次发生心跳停搏,是心脏骤停患者复苏后早期死亡的首要原因。约60%-70%成功复苏后的患者因出现低血压、休克或致命性室性心律失常而于24h内死亡,最终只有4.5%能存活出院。复苏后心功能不全主要归因于心脏骤停期间处于的全心缺血状态以及再灌注后所致的心肌损伤。促红细胞生成素(erythropoietin , Epo)是分子量为34KDa的糖蛋白类激素,主要作用于红系祖细胞,调节血循环中的红细胞容积。最近发现,Epo能在缺血再灌注中发挥广泛的组织保护作用,包括心脏、大脑、视网膜、肾脏、肝脏和皮肤等。在心肌缺血再灌注实验研究中, Epo能减少心肌梗死面积和减轻缺血再灌注损伤,改善心功能。基于缺血再灌注损伤是复苏后机体的主要病理生理过程。在心肺复苏期间给予Epo,有望发挥心肌保护作用,改善复苏后心功能。第一部分大鼠新型气管插管方法的建立 1.研究目的利用气管导管内置入直径为0.9mm的光纤作为光源,建立一种新型的大鼠经口气管插管方法。 2.研究方法将Sprague-Dawley大鼠分为2组(n=40):ⅰ)传统光源直视下的经口气管插管组(简称传统直视组n=20),ⅱ)气管导管内置入光纤引导下的经口气管插管组(简称光纤引导组n=20)。比较两组间的经口气管插管时间、插管次数、一次插管成功率。将上述经口气管插管的大鼠,与小型动物呼吸机连接,调整呼吸频率(60次/分)和潮气量(0.65ml/100g),持续通气60min后拔除气管插管,并比较拔除气管插管后1周的存活率。 3.结果 3. 1光纤引导组的经口气管插管时间和插管次数[(36.00±16.43)s和(1.05±0.22)],均较传统直视组明显减少([86.20±56.48)s和(1.75±1.02)],(P0.01)。 3. 2光纤引导组的经口气管插管一次成功率(95%)高于传统直视组(60%)(P0.05)。 3. 3光纤引导组拔除气管插管后的1周存活率(100%)高于传统直视组(80%);但其差异无统计学意义(P0.05)。 4.结论气管导管内光纤引导下的经口气管插管法是一种创新的大鼠气管插管方法,能实现快速、有效、准确的气管插管,值得推广应用。第二部分促红细胞生成素对心肺复苏后心肌的影响 1.研究目的观察窒息性大鼠心脏骤停-心肺复苏后的心功能和心肌损伤的情况;探讨促红细胞生成素对大鼠心肺复苏后的心肌保护作用。 2.研究方法夹闭气管8min ,建立窒息性大鼠心脏骤停-心肺复苏动物模型。Sprague-Dawley大鼠(n=24),随机分为3组:ⅰ)正常对照组(n=8):进行气管插管、血管置管,不予夹闭气管和心肺复苏。ⅱ)常规心肺复苏(CPR)组(n=8):夹闭气管8min后,进行胸外按压、机械通气和肾上腺素0.02mg/kg,于恢复自主循环(ROSC)后3min经股静脉推注0.9%NaCl 0.5ml。ⅲ)Epo治疗组(n=8):夹闭气管8min后,进行胸外按压、机械通气和肾上腺素0.02mg/kg,于ROSC后3min,将Epo5000U/kg与0.9%NaCl稀释成0.5ml后,注入股静脉。所有大鼠经右颈总动脉逆行插管至左心室和股动脉插管,持续监测心率(HR)、平均动脉压(MAP)、左室收缩压(LVSP)、左室舒张末压(LVEDP)、左室内压上升和下降最大变化速率(±dp/dtmax )和标准II导联心电图,肛门插入内置式温度探头记录深部直肠温度,用加热灯保持体温在36. 5±0. 5℃。于观察终点(ROSC后120min),采集血样测定血清CTnI含量,取出心脏,光镜和透射电镜观察心肌组织的损伤情况。 3.结果 3. 1三组间基线水平的HR、MAP、LVSP、LVEDP、±dp/dtmax无显著性差异(P 0.05)。 3. 2常规CPR组和Epo治疗组大鼠自主循环复苏后(ROSC)30min、60min、90min、120min的HR、MAP、LVSP和±dp/dtmax均正常对照组明显下降(P 0.01),ROSC120min的LVEDP较正常对照组明显升高(P 0.01)。 3. 3常规CPR组ROSC后120min血清CTnI水平较正常对照组明显升高(P 0.01);常规CPR组出现心肌细胞水肿变性、坏死、炎症细胞浸润,以及胞膜完整性丧失、线粒体肿胀、嵴断裂、溶解等心肌组织的损伤性改变。 3.4 Epo治疗组ROSC后30min、60min、90min、120min的LVSP(P0.05)、±dp/dtmax(P 0.01)较常规CPR组明显升高,ROSC120min的LVEDP较常规CPR组降低(P 0.01),两组间的HR和MAP无显著性差异(P 0.05)。 3.5同组内不同时间点比较显示,正常对照组的各项血流动力学指标无显著性差异(P 0.05),常规CPR组和Epo治疗组ROSC30 min、60min、90min、120min的HR、MAP、LVSP和±dp/dtmax较各自的基线水平明显下降(P0.01);ROSC120min的LVEDP较各自的基线水平显著升高(P0.01),而常规CPR组和Epo治疗组ROSC30 min、60min、90min、120min之间无显著性差异(P0.05)。 3.6 Epo治疗组与常规CPR组ROSC后120min血清CTnI水平比较无显著性差异(P 0.05);心肌组织的病理学结构和超微结构的损伤性改变较常规CPR组减轻。 4.结论 4.1窒息性大鼠心脏骤停-心肺复苏成功后存在心功能不全和心肌损伤。 4.2 Epo可以改善窒息性大鼠心脏骤停-心肺复苏成功后的心功能和减轻心肌损伤。全文结论 1.气管导管内光纤引导下的经口气管插管法是一种创新的大鼠气管插管方法,能实现快速、有效、准确的气管插管,值得推广应用。 2.窒息性大鼠心脏骤停-心肺复苏成功后存在心功能不全和心肌损伤。 3. Epo能改善窒息性大鼠心脏骤停-心肺复苏后的心功能和减轻心肌细胞损伤,对复苏后心肌具有保护作用。
[Abstract]:Background rats are commonly used experimental animals, but the tracheal intubation of rats has been a difficult problem because of small size, narrow oral cavity and great difficulty in exposing the glottis. The main methods of tracheal intubation in rats are the tracheotomy intubation method, blind insertion method and direct under intubation method. Therefore, it is necessary to seek a new intubation method to achieve safe and accurate endotracheal intubation in rats. A new method of endotracheal intubation in rats is established by using the endotracheal tube into a fiber with a diameter of 0.9mm as a light source and a guide wire.
Cardiac insufficiency after resuscitation is mainly characterized by hypotension, shock, fatal ventricular arrhythmias or heartbeat arrest, which is the primary cause of early death after the resuscitation of cardiac arrest patients. After the successful resuscitation of 60%-70%, patients died within 24h due to hypotension, shock or fatal ventricular arrhythmias, and only 4.5% can eventually be found. Survival after discharge. Cardiac dysfunction after resuscitation is mainly attributable to whole heart ischemia during cardiac arrest and myocardial injury induced by reperfusion.
Erythropoietin (Epo) is a glycoprotein hormone molecular weight 34KDa, which mainly acts on erythroid progenitor cells and regulates the volume of red cell in blood circulation. Recently, it has been found that Epo can play a wide tissue protection role in ischemia reperfusion, including heart, brain, retina, kidney, liver and skin. In the study of reperfusion, Epo can reduce infarct size, alleviate ischemia reperfusion injury and improve cardiac function.
Ischemia-reperfusion injury is the main pathophysiological process of the body after resuscitation, which is given to Epo during cardiopulmonary resuscitation, which is expected to play the role of myocardial protection and improve the cardiac function after resuscitation.
Part one establishment of new tracheal intubation in rats
1. purpose of research
A new type of rat trachea cannula was established by using a 0.9mm diameter optical fiber as the light source.
2. research methods
The Sprague-Dawley rats were divided into 2 groups (n=40): the oral tracheal intubation group (n=20) under the traditional light source (referred to as the traditional direct vision group), II) the endotracheal intubation group (referred to as the optical fiber guide group) under the optical fiber guided catheter (n=20). The time of intubation, the number of intubation and the success rate of the first intubation were compared. The rats of the tube intubation were connected with the miniature animal ventilator, the respiratory frequency (60 / sub) and the tidal volume (0.65ml/100g) were adjusted, and the tracheal intubation was extracted after continuous ventilation 60min, and the survival rate was compared for 1 weeks after the extraction of tracheal intubation.
3. results
3.1 the time of tracheal intubation and the number of intubation (36 + 16.43) s and (1.05 + 0.22) in the optical fiber guided group were significantly decreased ([86.20 + 56.48) s and (1.75 + 1.02)) compared with the traditional direct vision group (P0.01).
3.2 the success rate of fiberoptic catheterization group was 95% higher than that of the conventional group (60%) (P0.05).
3.3 the survival rate of 1 weeks after tracheal intubation in the fiber guided group was 100% higher than that in the conventional group (80%), but the difference was not statistically significant (P0.05).
4. conclusion
Endotracheal intubation under the fiber-optic guided endotracheal tube is an innovative method of endotracheal intubation in rats. It can achieve rapid, effective and accurate tracheal intubation. It is worth popularizing.
The second part is the effect of erythropoietin on myocardium after cardiopulmonary resuscitation.
1. purpose of research
To observe the cardiac function and myocardial injury after cardiac arrest and cardiopulmonary resuscitation in asphyxiated rats, and to explore the protective effect of erythropoietin on myocardium after cardiopulmonary resuscitation in rats.
2. research methods
.Sprague-Dawley rats (n=24) of asphyxiated rat cardiac arrest and cardiopulmonary resuscitation (n=24) were randomly divided into 3 groups: normal control group (n=8): tracheal intubation, vascular catheterization, no clipping trachea and cardiopulmonary resuscitation. II) routine cardiopulmonary resuscitation (CPR) group (n=8): after clamping the trachea 8min, chest compressions, mechanical ventilation were carried out. And adrenaline 0.02mg/kg, after the recovery of the autonomic circulation (ROSC), 3min was injected into the 0.9%NaCl 0.5ml. III Epo treatment group (n=8) through the femoral vein. After clamping the trachea 8min, the chest compressions, mechanical ventilation and adrenaline 0.02mg/kg were carried out after ROSC 3min. The femoral vein was injected into the femoral vein. All rats were reversed through the right cervical artery. Intubation of the left ventricle and femoral artery, continuous monitoring of heart rate (HR), mean arterial pressure (MAP), left ventricular systolic pressure (LVSP), left ventricular end diastolic pressure (LVEDP), the maximum change rate of left indoor pressure and decrease (+ dp/dtmax) and standard II lead electrocardiogram. The anus inserted into the built-in temperature probe to record the deep rectal temperature and keep the temperature with the heating lamp. At 36.5 + 0.5 degrees centigrade.
At the end of observation (120min after ROSC), blood samples were collected to measure serum CTnI level, and heart was removed. Light and transmission electron microscopy were used to observe the myocardial injury.
3. results
3.1 there was no significant difference in baseline HR, MAP, LVSP, LVEDP, and dp/dtmax between the three groups (P 0.05).
3.2 after the spontaneous circulation resuscitation (ROSC), 30min, 60min, 90min, 120min HR, MAP, LVSP and dp/dtmax in the normal control group were significantly decreased (P 0.01) in the routine CPR group and the Epo group (0.01).
3.3 the serum CTnI level of 120min after ROSC in the routine CPR group was significantly higher than that in the normal control group (P 0.01); in the routine CPR group, the myocardial cell edema degeneration, necrosis, inflammatory cell infiltration, and the loss of membrane integrity, mitochondrial swelling, crista fracture, dissolving and other myocardial tissue damage were changed.
In 3.4 Epo treatment group, LVSP (P0.05), 60min, 90min, 120min, LVSP (P0.05), and 120min 0.01 (P 0.01) in the treatment group were significantly higher than those in the conventional CPR group (0.01), and there was no significant difference between the two groups (0.05).
3.5 the comparison of different time points in the same group showed that there was no significant difference in the hemodynamic indexes of the normal control group (P 0.05). The HR, MAP, LVSP and + dp/dtmax of ROSC30 min, 60min, 90min, 120min in the routine CPR group and the Epo treatment group were significantly lower than those of the respective baseline levels. There was no significant difference in ROSC30 min, 60min, 90min and 120min between the conventional CPR group and the Epo treatment group (P0.05).
There was no significant difference in the level of CTnI between the 3.6 Epo treatment group and the routine CPR group (P 0.05) after ROSC (P 0.05), and the pathological structure and ultrastructure of the myocardial tissue were less damaged than those in the conventional CPR group.
4. conclusion
4.1 cardiac arrest in asphyxiated rats: cardiac dysfunction and myocardial injury after successful cardiopulmonary resuscitation.
4.2 Epo can improve cardiac function and reduce myocardial injury after asphyxiated cardiac arrest in rats.
Full text conclusion
1. endotracheal intubation under the guidance of fiber guided endotracheal tube is an innovative method of endotracheal intubation in rats. It can achieve rapid, effective and accurate tracheal intubation. It is worth popularizing.
2. cardiac arrest in asphyxiated rats: cardiac dysfunction and myocardial injury after successful cardiopulmonary resuscitation.
3. Epo can improve cardiac function and myocardial cell damage after asphyxiated cardiac arrest in rats, and has protective effect on myocardium after resuscitation.
【学位授予单位】：广州医学院
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R-332

【参考文献】