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研究生:吳欣榮
論文名稱:評估嗜中性球的功能--建立一套完整的嗜中性球功能分析系統
論文名稱(外文):Investigation of neutrophil function:establishment of comprehensive assays of neutrophil finction
指導教授:林亮音
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:醫事技術學研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2002
畢業學年度:90
語文別:中文
論文頁數:63
中文關鍵詞:嗜中性球功能
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白血球(leukocyte)是人體防禦系統的重要成員之一,其中又以嗜中性球(neutrophil)為防禦的第一道防線。當人體有外來物入侵時,嗜中性球接受到外來物或巨噬細胞所分泌的刺激分子(例如:lipopolysaccharide、Interleukin-8等),就會被活化,此時嗜中性球會先緊密地貼到內皮細胞(endothelial cell)表面,再穿過內皮細胞間隙到受感染組織,之後吞噬外來物並分泌一些殺菌物質(例如:超氧離子及過氧化氫等)將其去除。在這一連串的過程當中,若有任何一個步驟不能正常執行,嗜中性球便不能正常執行其防禦的功能。因此,建立臨床上評估嗜中性球之功能的方法是有其必要性的。
分析嗜中性球的標記如黏附分子(adhesion molecules) CD11b及CD18,可評估嗜中性球是否可以緊密地貼到內皮細胞上;分析其釋放殺菌物質過氧化氫的能力,可評估嗜中性球在吞噬了外來物之後,是否可以有效地消滅外來物;另外分析嗜中性球遷移(migration)的能力,可評估嗜中性球是否可以有效地穿過內皮細胞間隙,到達受感染的組織中。本研究係利用刺激物如:fMLP (N-formylmethionine- leucyl-phenylalanine)或PMA (Phorbol 12-Myristate 13-Acetate)刺激正常人的嗜中性球之後,用流式細胞儀偵測其CD11b、CD18及過氧化氫增加的比例。在CD11b及CD18方面,分別用5 nM及10 nM的fMLP刺激10分鐘之後,觀察它們增加的比例;而在過氧化氫方面,則是利用1 μM的PMA刺激10分鐘之後,觀察它增加的比例。另外在嗜中性球遷移的功能分析方面,用趨化功能測試組(transmigration assay),在100 nM fMLP趨化物的誘導下,計數正常人嗜中性球遷移到薄膜下的細胞增加比例。建立以上之評估嗜中性球功能的方法後,我們蒐集了27個正常人的嗜中性球,訂出了其刺激後所增加比例的正常範圍:CD11b的增加倍率為6.13 ± 1.90,CD18的增加倍率為2.53± 0.38,過氧化氫的增加倍率為252.96 ± 83.00,趨化實驗所增加的細胞倍率為1.62 ± 0.54。進一步比較3個CML病人嗜中性球的功能顯示,CML病人的嗜中性球經fMLP或PMA刺激之後,其CD11b、CD18及過氧化氫的增加比例雖然和正常人的增加比例沒有顯著的差異,但是和正常人比起來仍有偏低的趨勢。另外我們用一種針對CML不正常酪胺酸激脢的藥物STI571處理正常人的嗜中性球,再來偵測嗜中性球CD11b、CD18及過氧化氫的表現。實驗結果顯示,在藥物STI571處理過後,嗜中性球CD11b的表現量會有下降的趨勢,CD18的表現量會有上升的趨勢,而過氧化氫的表現量則沒有明顯的趨勢,這個實驗結果可以讓我們了解STI571對正常嗜中性球的影響。
在此,我們建立了一個快速、完整的嗜中性球功能分析系統,希望未來對臨床診斷有所幫助。

中文摘要..........................................1
英文摘要..........................................3
第一章 序論.......................................5
第二章 研究目的..................................16
第三章 材料與方法................................18
第四章 實驗結果..................................28
第五章 討論......................................35
參考資料.........................................40
圖表.............................................46

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