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  1. NTU Theses and Dissertations Repository
  2. 生物資源暨農學院
  3. 獸醫專業學院
  4. 臨床動物醫學研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23342
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor葉力森,張雅珮
dc.contributor.authorPen-Ting Liaoen
dc.contributor.author廖本庭zh_TW
dc.date.accessioned2021-06-08T04:59:27Z-
dc.date.copyright2011-08-22
dc.date.issued2011
dc.date.submitted2011-08-19
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1
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23342-
dc.description.abstract系統性低溫已經被證實對中風的病患具有治療效果,但許多併發症像是感染、心律不整以及凝血異常等等,往往增加致死率以及致病率,導致系統性低溫的治療效果大打折扣。隨著中風的疾
病生理機制以及對治療性低溫保護機制的日漸了解,局部治療性低溫的概念被提出及研究。本實驗的目的為利用大鼠的大腦局部性缺血模式,評估一個新發展出來的降溫裝置是否具有神經保護效果。
實驗使用十五隻體重300至400公克的Spraque-Dawley公鼠。本實驗依水流溫度分成14℃、26℃和無水流通過的對照組共三組,每組五隻大鼠。經開顱手術暴露出左側的近端中腦動脈後,利用雙極電燒器將其永久性阻斷。接著,將雙側總頸動脈暫時性阻斷三小時。恢復總頸動脈血流後半小時內,利用立體定位儀將降溫裝置放置於左前側大腦,靠近缺血核心處,並讓低溫水流經過裝置兩小時,接著立刻犧牲大鼠,並盡速將腦部組織取下,以腦切片盒將大腦切成厚度為2mm共八片之冠狀切片。再以0.05%,37℃的triphenyltetrazolium chloride (TTC)進行半小時染色。染色後,以10%福馬林固定至少24小時,並在48小時內利用數位相機記錄,之後利用電腦軟體分析。數位相機記錄後,隨機選每組三隻大鼠的腦切片進行石蠟包埋,再以Fluoro Jade C(FJC)、H&E染色進行組織病理分析。
TTC染色的結果顯示兩低溫組相較於控制組,在靠近缺血核心以及降溫裝置的前側大腦,都表現出顯著神經保護效果,但後側大腦只有26℃依舊表現顯著之神經保護效果。FJC染色的結果則和TTC有些許出入,FJC染色的結果顯示,在後側大腦同樣只有26℃表現顯著的神經保護效果,但前側大腦則兩降溫組皆無顯著神經保護效果。至於H&E染色則各組之間無顯著差異。根據上述結果,本次使用的降溫裝置當輸入水溫為26℃時,在相對的後側大腦,明確的表現出神經保護效果。因此認為,此降溫裝置具有未來發展潛力。
zh_TW
dc.description.abstractSystemic hypothermia has been considered beneficial to stroke patients. However, numerous complications ensued. The aim of this study is to evaluate the effect of a new invasive regional cooling device in cerebral ischemic injury in rat model.
A total of 15 adult male Spraque-Dawley rats, weighted 300-400 gram, were randomly assigned into three groups - cooling device applied at 14
en
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dc.description.tableofcontents目錄
口試委員會審定書 i
中文摘要 i
英文摘要 i
第一章 、序言 1
第一節 、前言與研究動機 1
第二節 、研究目的 1
第二章 、文獻探討 2
第一節 、大腦缺血性傷害的原因、病生理以及各式治療 2
第一項 、原因及風險因子 2
第二項 、腦缺血性傷害的病生理機制 3
第三項 、各式治療方法及效果 9
第二節 、治療性低溫於腦缺血性傷害的過往與現況 10
第一項 、治療性低溫的保護機制 10
第二項 、系統性治療性低溫 13
第四項 、局部性治療性低溫 17
第三節 、局部腦缺血性傷害大鼠動物模式-方法、優缺點 19
第三章 、實驗一 22
第一節 、實驗一之目的 22
第二節 、實驗一之材料及方法 22
第一項 、實驗設計 22
第二項 、實驗動物 23
第三項 、麻醉與監控 23
第四項 、放置降溫裝置及溫度計 23
第五項 、降溫裝置原理 24
第三節 、實驗一之結果 25
第四章 、實驗二 27
第一節 、實驗二之目的 27
第一節 、實驗二之材料及方法 27
第一項 、實驗設計 27
第二項 、實驗動物 27
第三項 、麻醉與監控 27
第四項 、局部腦部缺血 28
第五項 、放置降溫裝置 29
第六項 、動物犧牲及樣本處理 29
第七項 、組織病理分析 29
第八項 、結果分析 30
第九項 、統計分析 30
第三節 、實驗二之結果 30
第一項 、生理數值 30
第二項 、TTC染色結果 31
第二項 、灰階值比較 31
第三項 、H&E染色結果: 34
第四項 、Fluoro Jade C染色比較 40
第五章 、討論 43
第一節 、降溫裝置對腦部的降溫效率 43
第二節 、實驗二的溫度選擇 43
第三節 、使用TTC和Fluoro Jade C染色評估結果 44
第四項 、比較其他實驗結果 46
第五節 、實驗設計的限制 49
第一項 、犧牲時間點 49
第二項 、無行為評估 49
第六章 、結論 50
第七章 、參考資料 51

圖目錄
圖2-1、TOAST分類之特徵(源自Adams et al. Classification of subtype of acute ischemic stroke - definitions for use in a multicenter clinical-trial. Stroke. 1993;24:35-41) 2
圖2-1、簡述腦缺血性傷害病生理機制。 5
圖2-3、Unfolded protein response pathway (源自DeGracia DJ, Montie HL. Cerebral ischemia and the unfolded protein response. Journal of Neurochemistry. 2004;91:1-8) 6
圖2-4、HSPs被引發的機制。當細胞內蛋白質變性後,會和平時抑制HSPs的heat shock factors(HSFs)結合,使HSPs被釋放發揮功能,並進到細胞核內促使基因表現,進而增加。(源自 Yenari MA. Heat shock proteins and neuroprotection. In: Alzheimer C, ed. Molecular and cellular biology of neuroprotection in the cns. Kluwer Academic/Plenum Publishers and Landes Bioscience; 2002:281-300.) 7
圖2-5、Hsp70抗細胞凋零以及抗發炎機制。(源自Yenari MA, Liu J, Zheng Z, Vexler ZS, Lee JE, Giffard RG. Antiapoptotic and anti-inflammatory mechanisms of heat-shock protein protection. Ann Ny Acad Sci. 2005;1053:74-83) 8
圖2-6、大鼠利用血管內纖維形成局部缺血示意圖(源自 Kohno et al. A modified rat model of middle cerebral-artery thread occlusion under electrophysiological control for magnetic-resonance investigations. Magn Reson Imaging. 1995;13:65-71) 21
圖3-1、大鼠固定於立體定位實際照片,左上為俯照圖,右上為後照圖,左下為側照圖。固定經由門齒以及雙側耳道三點,固定整個頭部。 22
圖3-2、數位化溫度計。 24
圖3-3、降溫裝置與溫度計放置位置實際情況俯看圖。上方為鑽洞前,下方為鑽洞後。箭頭所指是bregma的位置,1、2、3代表洞1、洞2和洞3。 24
圖3-4、降溫裝置實際使用情形。箭號指的是放置於靠近降溫裝置進水口的溫度計,
箭頭代表水流方向。 25
圖3-5、降溫裝置原理以及循環水流示意圖,下方為點滴加壓器包覆於點滴袋外,上方為延長管及降溫裝置,下圖為降溫裝置,箭號代表水流方向。 25
圖3-6、降溫裝置於正常腦組織之降溫效率示意圖,橫軸除了最左邊為體溫,其他皆為輸入水溫。縱軸為腦部溫度。由左至右分別代表靜止腦溫、26、20、14和10℃輸入水溫得到的穩定降低腦溫。同一輸入水溫時,左邊直條為洞3位置紀錄之溫度,離洞1降溫裝置處約為3mm。右邊直條為洞2位置紀錄之溫度,離洞1降溫裝置處約為6mm。直條上方標示同樣英文字母時,代表為同質子集,也就是無顯著差異。 26
圖4-1、實驗二之實驗設計。 27
圖4-2、PE 50 tube合併6/0 尼龍縫線形成圈套(snare)阻斷總頸動脈(箭號)血流示意圖(左),以及實際情況(右)。 28
圖4-3、TTC染色結果,由左至右分別是control,14℃,26℃。 31
圖4-4、三組的每一片平均值以直條圖表現。直條圖上方字母相同,代表無統計上顯著差異。橫軸腦切片數字下方標計*,代表該組間有統計上顯著差異。組間具統計上顯著差異者為第2片、第3片以及第5片。 33
圖4-5、控制組第六片腦切片缺血交界處,可明顯發現圖中右側的缺血範圍內嗜伊紅性較弱。 35
圖4-6、H&E染色缺血交界處,上排為第3腦切片下排為第6腦切片,由左至右分別是控制組、14℃組和26℃組。圖片中下方為缺血範圍,可見交界處neuropil空泡化的現象更為明顯。 36
圖4-7、控制組H&E染色結果,第三片腦切片的缺血側是A和a ,正常側是B和b。第六腦切片的缺血側是C和c,正常側是D和d。可以看到缺血側的neuropil有明顯的空泡化以及perikarya明顯的濃縮。 37
圖4-8、14℃組H&E染色結果,第三片腦切片的缺血側是A和a ,正常側是B和b。第六腦切片的缺血側是C和c,正常側是D和d。可以看到缺血側的neuropil有明顯的空泡化以及perikarya明顯的濃縮。 38
圖4-9、26℃組H&E染色結果,第三片腦切片的缺血側是A和a ,正常側是B和b。第六腦切片的缺血側是C和c,正常側是D和d。可以看到缺血側的neuropil有明顯的空泡化以及perikarya明顯的濃縮。 39
圖4-10、Fluoro Jade C染色結果,左側為缺血側圖,右側為正常側對照圖。A為控制組前側,a為控制組後側;B為14組前側,14控制組後側;C為26組前側,c為26組後側。 41
圖4-11、Fluoro Jade C染色分析結果,橫軸為前後側分別,1為前側,2為後側,縱軸為缺血側平均值及正常側平均值之比值,數字越大代表陽性反應面積越大。直條上方同樣英文字母代表無統計上差異,為同質子集。可見前側14℃組明顯陽性反應。 42
圖5-1、使用pMCAO模式並於缺血開始前或缺血發生中開始降溫的實驗結果。(源自Krieger DW, Yenari MA. Therapeutic hypothermia for acute ischemic stroke: What do laboratory studies teach us? Stroke; a journal of cerebral circulation. 2004;35:1482-1489 46
圖 2-2、使用pMCAO模式並於缺血開始後開始降溫的實驗結果。(源自Krieger DW, Yenari MA. Therapeutic hypothermia for acute ischemic stroke: What do laboratory studies teach us? Stroke; a journal of cerebral circulation. 2004;35:1482-1489) 48
表目錄
表3-1、腦部溫度量測值的敘述性統計量表。 26
表4-1、三組生理數值平均值,P value>0.05時,各組之間無統計上顯著差異。 31
表4-2、三組切片1到切片8的mean GL/R的描述性統計量表。 32
表4-3、第1到第8片的mean GL/R 集合比較,p<0.05表示有統計上差異。 34
表4-4、第1到第5片的mean GL/R 集合比較,p<0.05表示有統計上差異。 34
表4-5、第6到第8片的mean GL/R 集合比較,p<0.05表示有統計上差異。 34
表4-6、第2、3片的Fluoro Jade C染色影像分析結果。 42
表4-7、第6、7片的Fluoro Jade C染色影像分析結果。 42
dc.language.isozh-TW
dc.subject局部性zh_TW
dc.subject局部永久性大腦缺血zh_TW
dc.subject系統性zh_TW
dc.subject中風zh_TW
dc.subject神經保護zh_TW
dc.subject治療性低溫zh_TW
dc.subjectstrokeen
dc.subjectneuroprotectionen
dc.subjectpermanent middle cerebral artery occlusionen
dc.subjectraten
dc.subjectselectiveen
dc.subjectregionalen
dc.subjectsystemicen
dc.subjecthypothermiaen
dc.title局部性低溫於大鼠永久性局部腦缺血模式之影響zh_TW
dc.titleThe effect of regional hypothermia in permanent focal cerebral ischemic injury in ratsen
dc.typeThesis
dc.date.schoolyear99-2
dc.description.degree碩士
dc.contributor.oralexamcommittee劉逸軒,林中天
dc.subject.keyword中風,局部永久性大腦缺血,治療性低溫,系統性,局部性,神經保護,zh_TW
dc.subject.keywordstroke,hypothermia,systemic,regional,selective,rat,permanent middle cerebral artery occlusion,neuroprotection,en
dc.relation.page62
dc.rights.note未授權
dc.date.accepted2011-08-19
dc.contributor.author-college獸醫專業學院zh_TW
dc.contributor.author-dept臨床動物醫學研究所zh_TW
顯示於系所單位:臨床動物醫學研究所

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