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研究生:李果濃
研究生(外文):Guo-Long Li
論文名稱:排骨靈活性成分AFB-1抑制人類嗜中性白血球釋放超氧自由基的機轉探討
論文名稱(外文):Inhibition of superoxide generation in human neutrophils by AFB-1 from Fissistigma bracteolatum
指導教授:黃聰龍黃聰龍引用關係
指導教授(外文):Tsong-Long Hwang
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:2005
畢業學年度:93
語文別:中文
論文頁數:83
中文關鍵詞:抗發炎去顆粒作用瓜馥木屬排骨靈人類嗜中性白血球呼吸爆破超氧自由基自瓜馥木屬
外文關鍵詞:6-hydroxy-578-trimethoxy-2-phenyl-chroman-4-one (AFB-1)anti-inflammatorycAMPERKelastaseFlavanoneformyl-L-methionyl
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中文摘要
嗜中性白血球扮演著宿主對抗細菌和黴菌的重要功能角色,不過,在一些特別的狀況下,嗜中性白血球過度或不適當活化反而會造成組織的損傷。目前已知,嗜中性白血球和許多疾病的形成,如類風濕性關節炎,缺血再灌流損傷,氣喘以及COPD (chronic obstructive pulmonary disease)等發炎疾病都有密切關係。在各種不同刺激劑的活化下,嗜中性白血球會釋放許多和發炎反應相關的物質,如超氧自由基(superoxide anion),顆粒性蛋白和生物活性脂質等。所以,如果能發展出可以調節嗜中性白血球功能的藥物將會是非常的重要。
為了進一步找尋新的抗發炎藥物,我們針對自瓜馥木屬植物 (Fissistigma sp.) 所分離出的15個純化物,對於人類嗜中性白血球在呼吸爆破及去顆粒的表現上進行研究。經由一連串的實驗發現,自排骨靈(Fissistigma bracteolatum)根莖所分離出的6-hydroxy-5,7,8-trimethoxy-2-phenyl-chroman-4-one (AFB-1)純化物對於由formyl-L-methionyl-L-leucyl-Lphenylalanine(fMLP)刺激活化人類嗜中性白血球所引起的超氧自由基的表現,具有最佳的抑制效果,其IC50 值經統計為0.34 ± 0.03M。相反地,AFB-1無法改變NADPH oxidase的活性表現,並且也不具有抗氧化的作用及清除自由基的能力。此外,AFB-1可抑制fMLP刺激活化人類嗜中性白血球所引起ERK1/2磷酸化的表現,但不影響p38的磷酸化。然而,AFB-1 對於PKC的活化劑所刺激誘導超氧自由基的釋放及fMLP所刺激誘導的去顆粒作用與Ca2+的調控,皆不具顯著性地抑制作用。而當給予PKA 抑制劑時,會明顯地逆轉AFB-1抑制fMLP刺激人類嗜中性白血球所引起的超氧自由基釋放作用。此外,AFB-1也確實會增加人類嗜中性白血球內cAMP的含量。綜合上述結果顯示:AFB-1抑制人類嗜中性白血球呼吸爆炸作用可能是經由cAMP及ERK1/2,而非經由Ca2+與p38的訊號路徑而來。
英文摘要
Neutrophils are known to play important roles in a host's defenses against invasion by microorganisms and in the pathogenesis of various diseases such as rheumatoid arthritis, ischemia-reperfusion injury, chronic obstructive pulmonary disease, and asthma. In response to diverse stimuli, activated neutrophils secrete a series of cytotoxins, such as the superoxide anion, a precursor of other reactive oxygen species, granule proteases, and bioactive lipids. Suppression of the extensive or inappropriate activation of neutrophils using drugs has been proposed as a way to ameliorate these inflammatory diseases. In a search for new anti-inflammatory agents, the effect of 15 pure compounds isolated from Fissistigma sp. on respiratory burst and degranulation in human neutrophils was tested. Among then, 6-hydroxy-5,7,8-trimethoxy-2-phenyl-chroman-4-one (AFB-1), isolated from the root of Fissistigma bracteolatum, showed the most-potent inhibitory effect on superoxide anion formation in formyl-L-methionyl-L-leucyl-Lphenylalanine (fMLP)-activated human neutrophils with an IC50 value of 0.34 ± 0.03 μM. In contrast, it failed to alter the subcellular NADPH oxidase activity, and it did not display antioxidant and superoxide anion-scavenging ability. Additionally, AFB-1 inhibited phosphorylation of ERK1/2 but not p38 MAPK in fMLP-induced human neutrophils. However, neither the protein kinase C activator–induced superoxide anion release nor the fMLP-stimulated elastase release and calcium mobilization was significantly inhibited by AFB-1. The inhibition of superoxide anion production in fMLP-stimulated human neutrophils by AFB-1 was reversed by protein kinase (PK)A inhibitors. Moreover, AFB-1 induced cAMP formation in human neutrophils. In summary, these results indicate that the inhibitory effect of AFB-1 on human neutrophil respiratory burst is through the cAMP- and ERK 1/2-dependent pathways, but not via the p38 and calcium-dependent pathways.
目錄

指導教授推薦書…………………................………...…………………...口試委員審定書………………………………………………………….
論文著作授權書…………………………………………………….…..Ⅰ
誌謝………………………………………………………………...…....Ⅱ
目錄..........................................................................................................Ⅲ縮寫表…………………………………………………………………....1
中文摘要…………...............………………...............…………………..3
英文摘要………...……….………………………………………………4
緒論………………………………………..…..........................................5
實驗材料與方法......................................................................................24
結果……………………………………………………………………..32
討論………………………………………………….…….....................37
圖表………………………………………………………......................42
參考文獻……………………………………………………..................62
參考文獻
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