臺灣博碩士論文加值系統

缺血性中風(ischemic stroke)為腦中風中最常見的一種，它致病原因是由於腦血管局部性阻塞，造成血液無法順利供給局部腦部組織，因而造成缺氧而導致腦組織受損或壞死。在缺血性中風發生後的初期，大腦組織會釋出許多發炎反應物質，其中有相當部分是來自磷脂質的代謝產物，這也表示脂質代謝物在缺血性中風的病理生理上扮演了一定的關鍵角色。過去利用基質輔助雷射脫離游離質譜法(MALDI mass spectrometry; MALDI-MS)觀察到缺血性中風後大鼠腦組織中大宗的磷脂醯膽鹼 (phosphatidylcholines; PCs)、溶血膽鹼磷(lysophosphatidylcholines; LPCs)、神經鞘磷脂(sphingomyelins; SMs)的分佈變化，並合併質譜影像(Mass Spectrometry Imaging; MSI)技術使這些脂質的增減與組織位置連結，但較少以此技術探討其他磷脂質(phospholipids; PLs)和神經鞘質(sphingolipids; SLs)的變化。此外，中風後血液中脂質變化與對應的腦組織脂質變化的比較，目前亦無系統性的探討。本研究將雄性Sprague-Dawley大鼠之中大腦動脈以手術方式造成永久梗塞(permanent middle cerebral artery occlusion; pMCAO)，24小時後，佐以適量之磷脂質內標物，將大鼠大腦皮質頂葉(parietal cortex)、顳葉(temporal cortex)、紋狀體(striatum)、及血漿內脂質含量以液相層析串聯質譜法(liquidchromatography-tandem mass spectrometry; LC/MS-MS)進行定量分析，並以中風取樣部位對側之正常腦組織質，及未中風大鼠血漿作為對照。除傳統的統計分析，結果亦以多變量分析(multivariate data analysis)來釐清不同解剖區域中區別中風與非中風狀態的PLs、SLs等脂類標誌物的組態。我們以此方式建立的高信賴度的脂質區辨模式，該模式的確認與實際的效應，則有待進一步的實驗室與臨床驗證。

Ischemic stroke is the most commonly encountered cerebrovascular accident. Blockage of a cerebral artery would interrupt the blood and oxygen supply to the affected arterial distributed territory and lead to various extent of parenchymal damage. The release of a plethora of lipid-derived inflammatory mediators precipitates the tissue responses following the circulatory interruption. Hence the phospholipid metabolism conceivably plays a critical role in mediating the pathophysiology of ischemic stroke. Most of matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS) studies of ischemic rat brain tissue reported the in situ changes of highly abundant phosphatidylcholines (PCs), lysophosphatidylcholines (LPCs), and sphingomyelins (SMs), and associated such changes with histology by MALDI-MS imaging. However, due to various reasons, changes in other classes of phospholipids (PLs) and sphingolipids (SLs) did not receive equal coverage in those studies. Nor were the ischemia-mediated lipid changes in brain parenchyma contrasted to, or compared with the changes in plasma. To address these concerns, the middle cerebral artery of male Sprague-Dawley rats were surgically occluded (permanent middle cerebral artery occlusion; pMCAO). The lipid levels in the ischemic temporal cortex, parietal cortex, striatum, and plasma were analyzed 24hrs after pMCAO using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the presence of internal standards, then contrasted the lipid levels in the respective contralateral non-ischemic regions as well as that in the plasma of sham rats. Other than the conventional statistical analyses, multivariate data analyses were also performed to identify the lipid species that distinguish the ischemic and non-ischemic states. High confidence lipidomic models were established based on these animal studies, while additional validation will further verify the practical utility of these models in the management of cerebral vascular events.

論文審定書 i
致謝 ii
摘要 iii
Abstract iv
目錄 vi
圖目錄 vii
表目錄 ix
字母縮寫表 x
前言 1
實驗規劃、材料和方法 6
結果與討論 12
結論 23
參考文獻 24
圖例 28
表格 40
附錄 54

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