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研究生:吳怡秀
研究生(外文):Wu, Yi-Hsiu
論文名稱:排骨靈活性成分bractelactone抑制嗜中性白血球超氧自由基和彈性蛋白酶釋出功能的機轉探討
論文名稱(外文):Inhibition of superoxide anion and elastase release in human neutrophils by bractelactone from Fissistigma bracteolatum
指導教授:黃聰龍黃聰龍引用關係
指導教授(外文):Hwang, Tsong-Long
學位類別:碩士
校院名稱:長庚大學
系所名稱:天然藥物研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:69
中文關鍵詞:超氧自由基彈性蛋白酶人類嗜中性白血球排骨靈
外文關鍵詞:superoxide anionelastasehuman neutrophilsbractelactoneFissistigma bracteolatum
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嗜中性白血球在宿主對抗微生物的防禦上是不可或缺的。瓜馥木屬排骨靈(Fissistigma bracteolatum),主要生長於中國南方和越南的攀援灌木,在傳統用藥上具有消炎和活血的功效。不過,排骨靈的藥理機轉仍然是未知的。由排骨靈的莖所分離出來的bractelactone (AFB-3),為天然物首次得到的chalcone衍生物。本論文使用人類嗜中性白血球的實驗模式來證明AFB-3的抗發炎作用。嗜中性白血球在風濕性關節炎、慢性肺阻塞疾病以及其他發炎疾病上扮演重要的角色,雖然如此,臨床上可用來直接調節嗜中性白血球引起的發炎症狀的藥物仍然很少。不過,AFB-3對嗜中性白血球的作用及機轉仍不清楚。AFB-3有抑制formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP)刺激嗜中性白血球釋出超氧自由基和elastase的作用,其IC50 分別為0.65 ± 0.06和0.85 ± 0.09 μM;AFB-3亦可抑制phorbol 12-myristate 13-acetate刺激嗜中性白血球所產生的超氧自由基。然而,這些抑制現象在protein kinase A抑制劑H89處理之下無法被逆轉;同樣地,AFB-3並不影響嗜中性白血球的cAMP生成量。另外,AFB-3不影響fMLP誘發的mitogen-activated protein kinase及Akt磷酸化現象。進一步研究指出,AFB-3抑制fMLP刺激的細胞外鈣進入,而非thapsigargin。再者,AFB-3本身具有直接抑制elastase活性的能力以及清除氮屬自由基的作用。綜上所述,AFB-3對於抑制fMLP刺激人類嗜中性白血球產生respiratory burst和去顆粒化的作用是經由減少鈣離子移動,以及本身具有直接抑制elastase活性的功能而來,而不是經由cAMP、PI3K、ERK和p38相關路徑。
Neutrophils are essential for host defense by phagocytosing and killing invading microorganisms. The Fissistigma bracteolatum, a climbing shrub, grows mainly in the southern part of China and Vietnam. As a folklore medicine, this plant is used to treat inflammation and enhance blood circulation. Bractelactone (AFB-3), a novel chalcone derivative, was isolated from the stems of Fissistigma bracteolatum. In this study, a cellular model in isolated human neutrophils, which are important in the pathogenesis of rheumatoid arthritis, chronic obstructive pulmonary disease, and other inflammatory diseases, was established to elucidate the anti-inflammatory functions of AFB-3. AFB-3 inhibited formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP)-induced superoxide anion (O2.-) generation and elastase release in human neutrophils in a concentration-dependent manner with IC50 values of 0.65 ± 0.06 and 0.85 ± 0.09 μM, respectively. AFB-3 also showed inhibition of O2.- generation in phorbol 12-myristate 13-acetate-activated human neutrophils. The inhibition of O2.- generation and elastase release in fMLP-stimulated human neutrophils by AFB-3 was not reversed by protein kinase A inhibitor H89. Consistent with this, AFB-3 did not alter cAMP levels in human neutrophils. Moreover, AFB-3 did not alter fMLP-induced phosphorylation of MAPKs and Akt. Significantly, AFB-3 concentration-dependently accelerated resequestration of cytosolic calcium in fMLP-activated human neutrophils. Further studies showed that AFB-3 inhibited extracellular calcium entry induced by fMLP but not thapsigargin. Additionally, AFB-3 could directly inhibit the activity of human neutrophil elastase and scavenge DPPH-radical in cell-free systems. In summary, these results indicate that the suppressive effects of AFB-3 on respiratory burst and degranulation of fMLP-induced human neutrophils are through the decrease of calcium mobilization and the direct inhibitory activity of elastase, but not via the cAMP, PI3K, ERK, and p38 dependent pathways.
縮寫表 1
中文摘要 4
英文摘要 5
研究動機 7
緒論 8
材料與方法 21
結果 26
討論 30
圖表 34
參考文獻 55
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中藥辭海第三卷,中國醫藥科技出版社,1997
全國中草藥滙編,人民衛生出版社,1996
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