緩慢間斷顱內(nèi)加壓法致豬腦死亡后炎癥反應及氧化應激對心肌的影響
發(fā)布時間：2018-10-17 12:59
【摘要】：目的：本研究旨在通過緩慢間斷顱內(nèi)加壓法建立豬腦死亡模型,觀察豬腦死亡模型建立過程中顱內(nèi)壓及腦電圖變化,與麻醉狀態(tài)下實驗動物進行比較,結合臨床表現(xiàn),完善實驗動物腦死亡判定標準,提高豬腦死亡判定準確率。檢測豬腦死亡和麻醉狀態(tài)下各時間段血清超氧化物歧化酶(SOD)、丙二醛(MDA)、白介素-6(IL-6)及單核細胞趨化蛋白-1(MCP-1)含量的變化。心肌組織檢查超氧化物歧化酶-1(SOD-1)和超氧化物歧化酶-2(SOD-2)的mRNA表達差異。心肌病理切片通過光學顯微鏡比較心肌細胞的變化情況。方法：6頭健康版納小耳豬隨機分為對照租(C組,3頭)和實驗組(E組,3頭)兩組。兩組均經(jīng)耳緣靜脈行全身麻醉后行氣管切開插管、呼吸機輔助呼吸、頸內(nèi)動靜脈插管和膀胱造瘺術。對照組在此基礎上僅通過呼吸、循環(huán)支持維持麻醉狀態(tài)10h；實驗組在此基礎上通過緩慢間斷顱內(nèi)加壓法建立腦死亡模型,即行顱骨鉆孔術并將Foley18F氣囊導管和顱內(nèi)壓監(jiān)測導線置于硬腦膜下腔,通過Foley氣囊導管緩慢向硬腦膜下腔內(nèi)注入約30-40ml生理鹽水以增加顱內(nèi)壓造成腦疝,建立腦死亡模型維持10h并持續(xù)監(jiān)測顱內(nèi)壓變化。實驗過程中詳細記錄用藥量、用藥時間、心率、心電圖、血壓、尿量及顱內(nèi)壓的變化情況。兩組實驗過程中除是否建立腦死亡模型外,其他實驗條件均相同。對照組在建模完成后分別于0.5、2、4、6、8、10h抽取靜脈血6ml用酶聯(lián)免疫吸附測定法(ELIAS)檢測血清超氧化物歧化酶、丙二醛、白介素-6、單核細胞趨化蛋白-1；實驗組在腦死亡模型建立后分別于0.5、2、4、6、8、10h抽取靜脈血16ml用酶聯(lián)免疫吸附測定法(ELIAS)檢測血清超氧化物歧化酶、丙二醛、白介素-6、單核細胞趨化蛋白-1；兩組實驗動物在維持10h后撤除呼吸、循環(huán)支持,開胸取心內(nèi)膜下心肌組織,檢測心肌超氧化物歧化酶-1(SOD-1)和超氧化物歧化酶-2(SOD-2) mRNA的表達情況和心肌細胞光學顯微鏡檢查。 結果： 1.對照組3頭實驗動物在麻醉狀態(tài)下通過呼吸、循環(huán)支持,全部存活10h,實驗成功率100%；實驗組3頭實驗動物的腦死亡模型建立成功率100%,術后10h存活率100%;共6頭豬用于課題研究。 2.血流動力學和強心藥物的用量：對照組在實驗過程中心功能未見明顯下降,有創(chuàng)動脈血壓檢測未見明顯變化,僅隨麻醉深度變化產(chǎn)生波動,未使用任何強心、血管活性藥物；實驗組在通過緩慢間斷顱內(nèi)加壓法建立豬腦死亡模型后,隨著心功能的逐漸下降,有創(chuàng)動脈血壓降低、心率下降,呈低血壓狀態(tài),需用強心、血管活性藥物如腎上腺素、多巴胺等才能維持基礎血壓,且實驗后期用藥量及用藥頻率較前期高。 3.血清IL-6：實驗組在腦死亡第2h后血清IL-6較對照組明顯升高,兩組對比有顯著性差異(P0.05)。 4.血清MCP-1：實驗組在腦死亡第2h后血清MCP-I較對照組明顯升高,兩組對比有顯著性差異(P0.05)。 5.血清SOD：實驗組在腦死亡后血清SOD較對照組明顯升高,兩組對比有顯著性差異(P0.05)。 6.血清MDA：實驗組在腦死亡后血清MDA較對照組明顯升高,兩組對比有顯著性差異(P0.05) 7.心肌組織SOD-1mRNA和SOD-2mRNA:實驗組較對照組心臟組織SODmRNA的表達均明顯升高,表達水平變化具有顯著性差異(P0.05)。 8.心肌組織光鏡：對照組未見明顯異常；實驗組可見心肌間質(zhì)水腫,心肌間質(zhì)血管收縮,炎細胞侵潤,局部可見心肌細胞溶解壞死。 結論： 1.本實驗應用緩慢間斷顱內(nèi)加壓法建立豬腦死亡模型,比較符合臨床腦死亡的發(fā)展過程,經(jīng)有效的呼吸、循環(huán)支持,腦死亡狀態(tài)可穩(wěn)定維持。 2.將動態(tài)腦電圖檢測、動態(tài)顱內(nèi)壓檢測應用于腦死亡模型建立的判定,在實驗中可行。 3.心功能下降是腦死亡后心臟重要的病理生理改變。兩組結果進行分析、比較,以非腦死亡狀態(tài)為參照,能更好的了解腦死亡狀態(tài)下心臟功能的改變情況及意義。 4.腦死亡狀態(tài)下,血清SOD、MDA、IL-6及MCP-1含量均有明顯升高。 5.腦死亡狀態(tài)下,心肌細胞內(nèi)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.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R654.2
本文編號：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.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R654.2
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