缓慢间断颅内加压法致猪脑死亡后炎症反应及氧化应激对心肌的影响
发布时间：2018-10-17 12:59
【摘要】：目的：本研究旨在通过缓慢间断颅内加压法建立猪脑死亡模型,观察猪脑死亡模型建立过程中颅内压及脑电图变化,与麻醉状态下实验动物进行比较,结合临床表现,完善实验动物脑死亡判定标准,提高猪脑死亡判定准确率。检测猪脑死亡和麻醉状态下各时间段血清超氧化物歧化酶(SOD)、丙二醛(MDA)、白介素-6(IL-6)及单核细胞趋化蛋白-1(MCP-1)含量的变化。心肌组织检查超氧化物歧化酶-1(SOD-1)和超氧化物歧化酶-2(SOD-2)的mRNA表达差异。心肌病理切片通过光学显微镜比较心肌细胞的变化情况。方法：6头健康版纳小耳猪随机分为对照租(C组,3头)和实验组(E组,3头)两组。两组均经耳缘静脉行全身麻醉后行气管切开插管、呼吸机辅助呼吸、颈内动静脉插管和膀胱造瘘术。对照组在此基础上仅通过呼吸、循环支持维持麻醉状态10h；实验组在此基础上通过缓慢间断颅内加压法建立脑死亡模型,即行颅骨钻孔术并将Foley18F气囊导管和颅内压监测导线置于硬脑膜下腔,通过Foley气囊导管缓慢向硬脑膜下腔内注入约30-40ml生理盐水以增加颅内压造成脑疝,建立脑死亡模型维持10h并持续监测颅内压变化。实验过程中详细记录用药量、用药时间、心率、心电图、血压、尿量及颅内压的变化情况。两组实验过程中除是否建立脑死亡模型外,其他实验条件均相同。对照组在建模完成后分别于0.5、2、4、6、8、10h抽取静脉血6ml用酶联免疫吸附测定法(ELIAS)检测血清超氧化物歧化酶、丙二醛、白介素-6、单核细胞趋化蛋白-1；实验组在脑死亡模型建立后分别于0.5、2、4、6、8、10h抽取静脉血16ml用酶联免疫吸附测定法(ELIAS)检测血清超氧化物歧化酶、丙二醛、白介素-6、单核细胞趋化蛋白-1；两组实验动物在维持10h后撤除呼吸、循环支持,开胸取心内膜下心肌组织,检测心肌超氧化物歧化酶-1(SOD-1)和超氧化物歧化酶-2(SOD-2) mRNA的表达情况和心肌细胞光学显微镜检查。 结果： 1.对照组3头实验动物在麻醉状态下通过呼吸、循环支持,全部存活10h,实验成功率100%；实验组3头实验动物的脑死亡模型建立成功率100%,术后10h存活率100%;共6头猪用于课题研究。 2.血流动力学和强心药物的用量：对照组在实验过程中心功能未见明显下降,有创动脉血压检测未见明显变化,仅随麻醉深度变化产生波动,未使用任何强心、血管活性药物；实验组在通过缓慢间断颅内加压法建立猪脑死亡模型后,随着心功能的逐渐下降,有创动脉血压降低、心率下降,呈低血压状态,需用强心、血管活性药物如肾上腺素、多巴胺等才能维持基础血压,且实验后期用药量及用药频率较前期高。 3.血清IL-6：实验组在脑死亡第2h后血清IL-6较对照组明显升高,两组对比有显著性差异(P0.05)。 4.血清MCP-1：实验组在脑死亡第2h后血清MCP-I较对照组明显升高,两组对比有显著性差异(P0.05)。 5.血清SOD：实验组在脑死亡后血清SOD较对照组明显升高,两组对比有显著性差异(P0.05)。 6.血清MDA：实验组在脑死亡后血清MDA较对照组明显升高,两组对比有显著性差异(P0.05) 7.心肌组织SOD-1mRNA和SOD-2mRNA:实验组较对照组心脏组织SODmRNA的表达均明显升高,表达水平变化具有显著性差异(P0.05)。 8.心肌组织光镜：对照组未见明显异常；实验组可见心肌间质水肿,心肌间质血管收缩,炎细胞侵润,局部可见心肌细胞溶解坏死。 结论： 1.本实验应用缓慢间断颅内加压法建立猪脑死亡模型,比较符合临床脑死亡的发展过程,经有效的呼吸、循环支持,脑死亡状态可稳定维持。 2.将动态脑电图检测、动态颅内压检测应用于脑死亡模型建立的判定,在实验中可行。 3.心功能下降是脑死亡后心脏重要的病理生理改变。两组结果进行分析、比较,以非脑死亡状态为参照,能更好的了解脑死亡状态下心脏功能的改变情况及意义。 4.脑死亡状态下,血清SOD、MDA、IL-6及MCP-1含量均有明显升高。 5.脑死亡状态下,心肌细胞内SOD-1mRNA、SOD-2mRNA含量均升高。
[Abstract]:Objective: The aim of this study was to establish the model of pig brain death by slow intermittent intracranial pressure method, observe the changes of intracranial pressure and EEG during the establishment of pig brain death model, compare with the experimental animals under the anesthesia state, combine the clinical manifestation, improve the experimental animal brain death judgment standard, improve that accuracy rate of the determination of the brain death of the pig. The changes of serum superoxide dismutase (SOD), (MDA), interleukin-6 (IL-6) and monocyte chemotaxis protein-1 (MCP-1) in pig brain death and anesthesia were detected. The mRNA expression of superoxide dismutase-1 (SOD-1) and superoxide dismutase-2 (SOD-2) was examined by myocardial tissue. The changes of cardiac myocytes were compared by optical microscope. Methods: Six healthy and small ear pigs were randomly divided into two groups: control group (group C, 3 head) and experimental group (group E, head 3). Both groups underwent tracheotomy intubation, ventilator-assisted respiration, internal jugular vein cannulation and cystostomy after systemic anesthesia. In the control group, the anesthesia state was maintained only by breathing and circulatory support, and the experimental group established the brain death model through the slow intermittent intracranial pressure method, namely, the skull drilling operation and the Foley18F balloon catheter and the intracranial pressure monitoring lead were placed in the subdural cavity. Approximately 30-40ml of physiological saline was injected slowly into the subdural cavity through the Foley balloon catheter to increase intracranial pressure to cause brain hernia, establish a brain death model for 10h and continuously monitor intracranial pressure changes. The changes of dosage, medication time, heart rate, electrocardiogram, blood pressure, heart rate and intracranial pressure were recorded in detail in the experiment. Except for the establishment of brain death model, the other experimental conditions were the same in both groups. In the control group, serum superoxide dismutase (SOD, MDA, IL-6, monocyte chemotaxis protein-1) were detected at 0. 5, 2, 4, 6, 8, and 10 h after the modeling was completed. In the experimental group, the serum superoxide dismutase (SOD, MDA, IL-6, monocyte chemotaxis protein-1) were detected with enzyme-linked immunosorbent assay (ELIAS) after the brain death model was established. The expression of superoxide dismutase-1 (SOD-1) and superoxide dismutase-2 (SOD-2) mRNA and optical microscope examination of cardiac myocytes were detected in the open chest. Results: 1. Three experimental animals in the control group were able to survive for 10h under the anesthesia state, the success rate was 100%, the success rate of brain death was 100%, the survival rate was 100% after operation, and 6 pigs were used in the experiment group. 2. Hemodynamics and dose of antiepileptic drugs: In the control group, there was no significant decrease in the function of the central function of the experimental process, and there was no obvious change in the blood pressure of invasive artery, but only with the change of the depth of anesthesia, and no strong use was used. Heart and blood vessel active drugs; in the experimental group, after the pig brain death model was established by the slow intermittent intracranial injection method, with the decrease of cardiac function, the invasive arterial blood pressure decreased, the heart rate was decreased, the blood pressure was low, and the blood vessels, vasoactive drugs such as epinephrine, dopamine, etc. were needed. the basal blood pressure can be maintained, and the dosage in the later period of the experiment and The serum IL-6 in the experimental group was significantly higher than that in the control group after the brain death, and the contrast between the two groups was significantly higher than that in the control group. The serum MCP-1 in the experimental group was significantly higher than that in the control group after brain death. Compared with the control group, the serum SOD in the experimental group was significantly higher than that in the control group. There was a significant difference between the two groups (P0.05). Serum MDA: the serum MDA in the experimental group after brain death was lower than that in the control group. The levels of SOD-1mRNA and SOD-2mRNA in myocardium were significantly higher than that in control group (P <0.05). There was a significant difference in the changes of expression level (P <0.05). 8. Myocardial tissue light microscope: no obvious abnormality was found in the control group. intermuscular stroma Conclusion: 1. The model of brain death of pig is established by slow intermittent intracranial pressure method in this experiment. The development of bed brain death can be stably maintained by effective breathing, circulatory support and brain death. Dynamic EEG detection and dynamic intracranial pressure detection applied to brain death model building 3. The decrease of cardiac function is the important pathophysiological change of the heart after brain death. Compared with the non-brain death status, it can better understand the changes of cardiac function in brain death and its significance.. 4. The levels of SOD, MDA, IL-6 and MCP-1 in serum were significant in the state of brain death.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R654.2
本文编号：2276730
[Abstract]:Objective: The aim of this study was to establish the model of pig brain death by slow intermittent intracranial pressure method, observe the changes of intracranial pressure and EEG during the establishment of pig brain death model, compare with the experimental animals under the anesthesia state, combine the clinical manifestation, improve the experimental animal brain death judgment standard, improve that accuracy rate of the determination of the brain death of the pig. The changes of serum superoxide dismutase (SOD), (MDA), interleukin-6 (IL-6) and monocyte chemotaxis protein-1 (MCP-1) in pig brain death and anesthesia were detected. The mRNA expression of superoxide dismutase-1 (SOD-1) and superoxide dismutase-2 (SOD-2) was examined by myocardial tissue. The changes of cardiac myocytes were compared by optical microscope. Methods: Six healthy and small ear pigs were randomly divided into two groups: control group (group C, 3 head) and experimental group (group E, head 3). Both groups underwent tracheotomy intubation, ventilator-assisted respiration, internal jugular vein cannulation and cystostomy after systemic anesthesia. In the control group, the anesthesia state was maintained only by breathing and circulatory support, and the experimental group established the brain death model through the slow intermittent intracranial pressure method, namely, the skull drilling operation and the Foley18F balloon catheter and the intracranial pressure monitoring lead were placed in the subdural cavity. Approximately 30-40ml of physiological saline was injected slowly into the subdural cavity through the Foley balloon catheter to increase intracranial pressure to cause brain hernia, establish a brain death model for 10h and continuously monitor intracranial pressure changes. The changes of dosage, medication time, heart rate, electrocardiogram, blood pressure, heart rate and intracranial pressure were recorded in detail in the experiment. Except for the establishment of brain death model, the other experimental conditions were the same in both groups. In the control group, serum superoxide dismutase (SOD, MDA, IL-6, monocyte chemotaxis protein-1) were detected at 0. 5, 2, 4, 6, 8, and 10 h after the modeling was completed. In the experimental group, the serum superoxide dismutase (SOD, MDA, IL-6, monocyte chemotaxis protein-1) were detected with enzyme-linked immunosorbent assay (ELIAS) after the brain death model was established. The expression of superoxide dismutase-1 (SOD-1) and superoxide dismutase-2 (SOD-2) mRNA and optical microscope examination of cardiac myocytes were detected in the open chest. Results: 1. Three experimental animals in the control group were able to survive for 10h under the anesthesia state, the success rate was 100%, the success rate of brain death was 100%, the survival rate was 100% after operation, and 6 pigs were used in the experiment group. 2. Hemodynamics and dose of antiepileptic drugs: In the control group, there was no significant decrease in the function of the central function of the experimental process, and there was no obvious change in the blood pressure of invasive artery, but only with the change of the depth of anesthesia, and no strong use was used. Heart and blood vessel active drugs; in the experimental group, after the pig brain death model was established by the slow intermittent intracranial injection method, with the decrease of cardiac function, the invasive arterial blood pressure decreased, the heart rate was decreased, the blood pressure was low, and the blood vessels, vasoactive drugs such as epinephrine, dopamine, etc. were needed. the basal blood pressure can be maintained, and the dosage in the later period of the experiment and The serum IL-6 in the experimental group was significantly higher than that in the control group after the brain death, and the contrast between the two groups was significantly higher than that in the control group. The serum MCP-1 in the experimental group was significantly higher than that in the control group after brain death. Compared with the control group, the serum SOD in the experimental group was significantly higher than that in the control group. There was a significant difference between the two groups (P0.05). Serum MDA: the serum MDA in the experimental group after brain death was lower than that in the control group. The levels of SOD-1mRNA and SOD-2mRNA in myocardium were significantly higher than that in control group (P <0.05). There was a significant difference in the changes of expression level (P <0.05). 8. Myocardial tissue light microscope: no obvious abnormality was found in the control group. intermuscular stroma Conclusion: 1. The model of brain death of pig is established by slow intermittent intracranial pressure method in this experiment. The development of bed brain death can be stably maintained by effective breathing, circulatory support and brain death. Dynamic EEG detection and dynamic intracranial pressure detection applied to brain death model building 3. The decrease of cardiac function is the important pathophysiological change of the heart after brain death. Compared with the non-brain death status, it can better understand the changes of cardiac function in brain death and its significance.. 4. The levels of SOD, MDA, IL-6 and MCP-1 in serum were significant in the state of brain death.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R654.2
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