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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林靖愉(Ching-Yu Lin) | |
dc.contributor.author | Chih-Hsien Lin | en |
dc.contributor.author | 林芷嫺 | zh_TW |
dc.date.accessioned | 2021-06-15T06:00:53Z | - |
dc.date.available | 2014-09-09 | |
dc.date.copyright | 2010-09-09 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-08-17 | |
dc.identifier.citation | Berliner, J. A., Mohamad, N., Alan, M., Judith, A. B., et al. (1995). Atherosclerosis: basic mechanisms. Oxidation, inflammation, and genetics. Circulation, 91: 2488-2496.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47461 | - |
dc.description.abstract | 許多流行病學研究都已證實細懸浮微粒(PM2.5)會對心血管系統產生負面的健康效應,尤其對本身已罹患心血管疾病的病人會產生更嚴重的反應。大氣中懸浮微粒對心血管系統的致病機制有直接作用和間接作用兩個假說,其中心血管系統的損傷都與體內脂質的平衡擾亂有很大的相關性;磷脂醯膽鹼類,在磷脂質內佔最大比例的脂質,可能與懸浮微粒暴露誘導心血管系統異常有密切的關係。本研究比較心肌受損大鼠暴露到濃縮大氣懸浮微粒後血漿內脂質,尤其是磷脂醯膽鹼類的變異,希望能觀察到濃縮大氣懸浮微粒對易感受族群的分子層次影響。
雄性SD大鼠注射isoproterenol使其發展成為心肌受損大鼠後,會暴露65天濃縮大氣細懸浮微粒(平均濃度149 µg/m3)再進行犧牲,取大鼠血漿樣本,以Folch萃取法萃取脂溶性代謝物並使用核磁共振儀與質譜儀進行分析,得到的圖譜以多變相檢定法觀察暴露組與控制組血漿內脂溶性代謝物是否有差異。 統計結果顯示心肌受損大鼠暴露到濃縮大氣細懸浮微粒後,血漿內的脂質含量出現變動,核磁共振圖譜可發現暴露組的總脂肪酸鏈雙鍵數顯著下降,細懸浮微粒造成生物體內氧化壓力提升可能是雙鍵數下降的一個原因;質譜儀的資料則顯示單碳鏈的磷脂醯膽鹼類在暴露組會有一致的下降趨勢,可能是血小板凝結因子大量消耗,促使單碳鏈磷脂醯膽鹼類以特殊途徑生成血小板凝結因子而降低在血漿中的濃度;某些雙碳鏈的磷脂醯膽鹼類則會提高在血漿中的濃度。找出這些在血漿內出現變動的脂質與磷脂醯膽鹼類,可以提供後續研究一些線索與方向並縮小分析範圍,針對這幾個物質進行更進一步的研究,以解釋濃縮大氣細懸浮微粒的可能致病機制並發掘可能的生物標誌以做為前期預防的指標。 | zh_TW |
dc.description.abstract | Many epidemiologic studies have demonstrated that fine particles (PM2.5) will produce cardiovascular adverse effects, especially for those who already have cardiovascular disease. There have been two hypotheses about the mechanisms of ambient particulate in cardiovascular system. The first one is through direct effect. PM or its’ soluble materials activate the neural reflexes and disarray the heart rate rhythm, which sequentially conducting the cardiac arrhythmias. The other one is the indirect effect, which caused by increased oxidative stress and inflammation, then inducing the thrombosis and the atherosclerosis. Either direct effects or indirect effects will disturb the lipids’ balance during the course of cardiovascular disease. Glycerophosphocholine (PC), which accounted the most ratios in phospholipids, are likely to be affected by PM2.5 exposure and cause cardiovascular abnormal. In our study, we compare the differences of PC in myocardial rat plasma between PM2.5 exposure and control group. We intend to examine the molecular events of concentrated ambient particles in plasma of susceptible population.
Male SD rats were exposed to PM2.5 for 65 days after they became myocardial damaging rats by isoproterenol(150 mg/kg). After animals were scarified, plasma samples were taken and snap frozen. The samples were extracted by Folch’s extraction method and the hydrophobic layer was removed for farther analysis. Both NMR and MS were used to analyze the hydrophobic metabolite extracts of the rat plasma. The data from NMR and MS were pretreated for multivariate analysis to examine the hydrophobic metabolic variation among samples. The statistic results demonstrated lipid fluctuations in rat plasma after PM2.5 exposure. NMR data showed that fatty acyl chain -CH2CH= and =CHCH2CH= decreased in exposure group. The raised oxidative stress after PM2.5 exposure may explain the decline of unstaurated fatty acyl chain. MS data indicated that the concentrations of lysoPCs were declined in exposure group. Moreover, numerous diacyl-phosphatidylcholine and alkyl-acyl-phosphatidylcholine were increased in the plasma of PM2.5 exposed rats. Understanding the fluctuations of PC in plasma can help us to elucidate the possible mechanism of PM2.5 toxicity and to mine the possible biomarkers for cardiovascular disease progress. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T06:00:53Z (GMT). No. of bitstreams: 1 ntu-99-R97844012-1.pdf: 3006308 bytes, checksum: b9f5b06c406c2d20570581c4459126f2 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 國立臺灣大學碩士學位論文 口試委員會審定書 I
誌謝 II 摘要 III ABSTRACT IV 目錄 V 圖目錄 VIII 表目錄 X 第一章 前言 1 1.1 研究背景 1 1.2 研究目的 2 第二章 文獻回顧 3 2.1 濃縮大氣懸浮微粒 3 2.1.1 簡介 3 2.1.2 懸浮微粒的流行病學研究 5 2.1.3 懸浮微粒的心血管病理研究 7 2.2 心血管疾病 8 2.2.1 簡介 8 2.2.2 心血管疾病之危險因子 9 2.2.3 後天心血管疾病致病過程 10 2.3 代謝體學 11 2.3.1 代謝體學 11 2.3.2 代謝物偵測方法 12 2.3.3 數據處理 14 2.4 脂質體學 15 2.4.1 簡介 15 2.4.2 脂質分類及命名法 16 2.4.3 磷脂醯膽鹼類和磷脂醯乙醇胺類 18 第三章 實驗材料與方法 20 3.1 實驗架構 20 3.2 濃縮大氣懸浮微粒暴露實驗 21 3.2.1 細粒徑微粒濃縮器(Ultrafine Particle Concentrator, UFPC) 21 3.2.2 實驗動物 22 3.2.3 暴露系統(exposure system) 22 3.2.4 濃縮大氣懸浮微粒暴露 23 3.3脂質標準品配置 24 3.3.1 脂質標準品 24 3.3.2 有機溶劑與實際樣本內標準品配置 25 3.4樣本前處理 25 3.5核磁共振儀分析 27 3.5.1 水溶性代謝物 28 3.5.2 脂溶性代謝物 28 3.5.3 一維1H核磁共振實驗 29 3.5.4 二維J-resolved核磁共振實驗 29 3.6核磁共振圖譜資料處理 30 3.6.1 圖譜處理 30 3.6.2 統計分析 31 3.6.3 代謝物鑑定 33 3.7超效能液相層析儀/質譜儀 33 3.7.1 超效能液相層析儀 33 3.7.2 質譜儀 34 3.8超效能液相層析儀/質譜儀之品保/品管 35 3.8.1超效能液相層析儀/質譜儀檢量線之建立 35 3.8.2 品保品管 36 3.9超效能液相層析儀/質譜儀資料分析 36 3.9.1 磷脂醯膽鹼類的定性方法 36 3.9.2 統計分析 37 第四章 結果 39 4.1 樣本萃取方法比較 39 4.1.1 水溶性代謝物 39 4.1.2 脂溶性代謝物 40 4.1.3 Folch萃取法樣本回收率 41 4.2 超效能液相層析儀/質譜儀條件建立 42 4.2.1 磷脂醯膽鹼類與磷脂醯乙醇胺類的定性 42 4.2.2 質譜儀器參數最適化 43 4.2.3 BEH C18管柱層析條件最適化 44 4.2.4 檢量線之建立 45 4.3 濃縮懸浮大氣微粒暴露大鼠的血漿分析 46 4.3.1 核磁共振分析結果 46 4.3.2 液相層析儀/質譜儀分析結果 49 第五章 討論 51 5.1 萃取方法比較 51 5.2 液相層析/質譜儀方法建立 52 5.2.1 磷脂醯膽鹼類與磷脂醯乙醇胺類的定性 52 5.2.2 超效能液相層析儀/質譜儀參數最適化 54 5.2.3 方法驗證 55 5.3 濃縮大氣懸浮微粒暴露實驗 55 5.3.1 核磁共振儀分析結果 55 5.3.2 超效能液相層析儀/質譜儀分析結果 56 5.3.3 核磁共振儀與超效能液相層析儀/質譜儀結果比較 57 5.4 研究限制 58 5.5 結論 59 參考文獻 60 圖目錄 圖一 國際純粹與應用化學聯合會(IUPAC)對脂質之八大分類及其個別結構 73 圖二 六大類磷脂質頭基之結構圖 74 圖三 三種碳鏈結構及其與磷脂質之鍵結位置 75 圖四 虛擬衝擊法之細粒徑微粒濃縮器濃縮流程圖 76 圖五 暴露期間內濃縮微粒重量濃度 77 圖六 磷脂醯膽鹼類鑑定方法 78 圖七 萃取大鼠血漿水溶性代謝物之1H NMR圖譜 79 圖八 萃取大鼠血漿水溶性代謝物之二維 J-resolved NMR圖譜 80 圖九 萃取大鼠血漿水溶性代謝物之二維J-resolved NMR實驗所投射的一維圖譜 81 圖十 Foltch和Laaksonen萃取方法萃取水溶性代謝物的1H NMR圖譜之主成分分析圖 82 圖十一 萃取大鼠血漿脂溶性代謝物之1H NMR圖譜 83 圖十二 萃取大鼠血漿脂溶性代謝物之二維J-resolved NMR圖譜 84 圖十三 萃取大鼠血漿脂溶性代謝物之二維J-resolved NMR實驗所投射的一維圖譜 85 圖十四 Foltch和Laaksonen萃取方法萃取脂溶性代謝物的1H NMR圖譜之主成分分析圖 86 圖十五 不同有機溶劑(甲醇或乙腈)各別添加不同鹽類(醋酸胺或醋酸胺+甲酸)之層析比較圖 87 圖十六 四種有機相(甲醇、乙腈、乙腈/異丙醇(5/2)和乙腈/異丙醇(5/2)添加鹽類)對不同磷脂醯膽鹼類及磷脂醯乙醇胺類脂層析波峰與訊號強度比較圖 88 圖十七 暴露濃縮大氣懸浮微粒大鼠血漿脂溶性代謝物之1H NMR圖譜 89 圖十八 暴露濃縮大氣懸浮微粒大鼠血漿脂溶性代謝物之二維J-resolved NMR圖譜 90 圖十九 暴露濃縮大氣懸浮微粒大鼠血漿脂溶性代謝物之二維J-resolved NMR實驗所投射的一維圖譜 91 圖二十暴露濃縮大氣懸浮微粒大鼠血漿脂溶性代謝物的J-resolved NMR圖譜之主成分分析圖 92 圖二十一 暴露濃縮大氣懸浮微粒大鼠血漿脂溶性代謝物的 J-resolved NMR圖譜偏最小平方法辨別分析圖 93 圖二十二 大鼠血漿脂溶性代謝物第一潛在變數負荷量 94 圖二十三 利用超效能液相層析儀/質譜儀分析暴露濃縮大氣懸浮微粒大鼠血漿內磷脂醯膽鹼類得到之一維層析圖譜 95 圖二十四 利用超效能液相層析儀/質譜儀分析暴露濃縮大氣懸浮微粒大鼠血漿內磷脂醯膽鹼類之二維圖譜 96 圖二十五 暴露濃縮大氣懸浮微粒大鼠血漿內磷脂醯膽鹼類質譜分析之主成分分析圖 97 圖二十六 暴露濃縮大氣懸浮微粒大鼠血漿內磷脂醯膽鹼類質譜分析之第二主成分負荷量圖 98 圖二十七 磷脂醯膽鹼類第二主成分負荷量圖中VIP值大於1的磷脂醯膽鹼類 99 表目錄 表一. 人體血漿中脂蛋白的分類與組成 100 表二. 脂肪酸之名稱、化學結構與表示法 101 表三. 標準品結構圖 102 表四. 脂溶性代謝物圖譜去除水或氯仿化學位移範圍 103 表五. 1H NMR圖譜以內標校正後總面積比例 104 表六. Folch萃取法在不同濃度標準品的萃取率 105 表七 質譜中磷脂醯膽鹼類之離子形態相對百分比 106 表八 磷脂醯膽鹼類/磷脂醯乙醇胺類最佳撞擊能量和進樣錐電壓107 表九 質譜儀最佳參數 108 表十. UPLC BEH C18 管柱在不同基質的檢量線 109 表十一. 暴露濃縮大氣懸浮微粒大鼠血漿脂溶性代謝物之核磁共振儀鑑定 111 表十二. 暴露濃縮大氣懸浮微粒大鼠血漿之脂溶性代謝物相對含量NMR分析結果 112 表十三. UPLC/MS/MS分析大鼠血漿內磷脂醯膽鹼類 113 表十四. 暴露濃縮大氣懸浮微粒大鼠血漿之磷脂醯膽鹼類相對含量MS分析結果 115 | |
dc.language.iso | zh-TW | |
dc.title | 利用心肌損傷大鼠探討濃縮大氣懸浮微粒對血漿磷脂醯膽鹼類的影響 | zh_TW |
dc.title | Effects of concentrated ambient fine particles on plasma glycerophosphocholine in a myocardial rat model | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 鄭美玲(Mei-Ling Cheng),鄭尊仁(Tsun-Jen Cheng),陳家揚(Chia-Yang Chen) | |
dc.subject.keyword | 懸浮微粒,心血管疾病,脂質學,磷脂醯膽鹼類,核磁共振儀,質譜儀, | zh_TW |
dc.subject.keyword | particulate matter,cardiovascular disease,lipidomics,phosphatidylcholine (PC),nuclear magnetic resonance (NMR),mass spectrometry (MS), | en |
dc.relation.page | 115 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-08-17 | |
dc.contributor.author-college | 公共衛生學院 | zh_TW |
dc.contributor.author-dept | 環境衛生研究所 | zh_TW |
顯示於系所單位: | 環境衛生研究所 |
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