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論文名稱 Title |
以基質輔助雷射游離質譜法研究唾液中的防禦素的分佈及其與生長的關係 Investigation of Human Neutrophil Peptide in Saliva and Their Relationship with Growth by Matrix-Assisted Laser Desorption Ionization/Time-of-Flight Mass Spectrometry |
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系所名稱 Department |
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畢業學年期 Year, semester |
語文別 Language |
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學位類別 Degree |
頁數 Number of pages |
87 |
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研究生 Author |
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指導教授 Advisor |
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召集委員 Convenor |
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口試委員 Advisory Committee |
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口試日期 Date of Exam |
2009-06-27 |
繳交日期 Date of Submission |
2009-06-30 |
關鍵字 Keywords |
防禦素、唾液、基質輔助雷射脫附 saliva, HNPs, MALDI-TOF MS |
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統計 Statistics |
本論文已被瀏覽 5645 次,被下載 3545 次 The thesis/dissertation has been browsed 5645 times, has been downloaded 3545 times. |
中文摘要 |
唾液在口腔中扮演重要的角色,因為其含有碳酸鹽、磷酸鹽和蛋白質,對牙齒有保護作用,而含有的澱粉酶能夠幫助消化。唾液主要由耳下腺、舌下腺和頜下腺這三對唾液腺共同分泌;唾液分泌的質與量,會受到大腦皮層的控制,也會受到飲食、環境、年齡以及情緒或病變等影響。而唾液中的防禦素與人體的健康有著密切的關係;此研究探討防禦素(Human neutrophil peptides, HNPs)與年齡的關係,因HNPs 為抗菌成份的一種,而根據文獻, HNPs 是具有抵抗細菌、黴菌和病毒的能力。唾液是人體體液中易於取得且其取樣方法不具侵入性,在此研究中我們開發一快速偵測的技術來觀察不同年齡層中由耳下腺與舌下腺/頷下腺所分泌的唾液其 HNPs 分佈的情形。 本研究利用基質輔助雷射游離質譜法(MALDI-TOF MS)直接分析唾液,此方法具有樣品需要量少、分析速度快、靈敏度高、鹽類容許度大等優點。本實驗使用吸附的方式取樣後,以芥子酸(sinapinic acid, SA)當作基質直接以 MALDI-TOF MS 進行偵測。 取0.5-84歲共70人之耳下腺與舌下腺/頷下腺唾液為樣品,每10歲為一範圍共7個範圍,每個年齡層以10名自願者當作代表。由結果中可知,耳下腺的唾液樣品有99 % 可以被偵測到HNPs的訊號,但在舌下腺/頷下腺30歲以下樣品中並沒有偵測到HNPs的訊號,且30歲以上舌下腺/頷下腺樣品偵測到 HNPs 的訊號之比例隨年齡逐漸上升,尤其在 60 歲以上的樣品中, 90 % 都偵測到 HNPs的訊號。此外,我們也利用不同方式,像是洗牙、味覺條件改變,研究誘導舌下腺/頷下腺中 HNPs 是否產生;也發現 HNP 1-3 的表現與家族基因有密切的關係。此外,我們也使用ELISA來測量唾液中耳下腺與舌下腺/頷下腺中 HNP 1-3 的總量,可看出舌下腺/頷下腺含 HNP 1-3 的比例也是隨著年齡而逐漸上升,因此此技術可以用於快速偵測不同年齡層唾液中 HNPs 的分佈差異與其在唾液中含量的關係。 |
Abstract |
none |
目次 Table of Contents |
摘要........................................................................................ I 目錄........................................................................................III 圖目錄................................................................................V 表目錄.............................................................................X 壹、 緒論............................................................1 一、 前言...................................................................1 二、 基質輔助雷射脫附游離質譜法(MALDI).....3 三、 酵素連結免疫吸附法(ELISA).................12 四、 口腔功能.......................................................15 五、 唾液腺簡介...................................................17 六、 防禦素簡介...................................................21 七、 論文目標........................................................27 貳、 實驗................................................................29 一、 儀器設備...................................................................29 二、 試劑配製及樣品.......................................................30 三、 實驗流程...................................................................31 參、 結果與討論...............................................................36 肆、 結論...........................................................................68 伍、 參考資料...................................................................69 |
參考文獻 References |
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