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煙麴黴活性成分 FM-8 抑制人類嗜中性白血球發炎機轉之探討

為了解決safari park kanchana的問題，作者郭昱廷 這樣論述：

指導教授推薦書論文口試委會員審定書誌謝 iii中文摘要 ivAbstract v目錄 vi表目錄 ix圖目錄 x縮寫表 xii第一章 緒論 11.1 研究動機 11.2 麴菌屬真菌 Aspergillus species 11.3 FM-8 介紹 21.4 免疫系統相關疾病 41.5 嗜中性白血球簡介 41.5.1 呼吸爆破作用 (Respiratory burst) 61.5.2 去顆粒化作用 (Degranulation) 61.6 嗜中性白血球導致之相關疾病 71.7 甲醯化胜肽 (N-formyl

peptides) 81.8 甲醯化胜肽受體 (Formyl peptide receptors, FPRs) 91.9 嗜中性白血球內免疫調控相關之訊息傳遞路徑 111.9.1 G protein-couple receptors (GPCRs) 111.9.2 Akt 121.9.3 Mitogen-activated protein kinases (MAPKs) 121.9.4 細胞內Ca2+對嗜中性白血球的影響 131.10 細胞週期蛋白依賴型激酶 (Cyclin-dependent kinases, CDKs) 131.11 細胞凋

亡 (Apoptosis) 141.12 活性氧類 (Reactive oxygen species, ROS) 14第二章 實驗材料與方法 172.1 實驗材料 172.2 實驗方法 172.2.1 人類嗜中性白血球製備 172.2.2 細胞培養與分化 182.2.3 細胞轉染 182.2.4 細胞存活率 182.2.5 活性氧自由基釋放之測定 192.2.6 彈性蛋白酶釋放之測定 202.2.7 測定直接清除自由基活性 202.2.8 彈性蛋白酶活性之測定 222.2.9 細胞毒性測試 232.2.10 細

胞內 Ca2+ 濃度測定 232.2.11 細胞表面 CD11b 的表現 242.2.12 趨化作用測試 242.2.13 接受體結合測定 242.2.14 西方墨點法 252.2.15 細胞群落篩選 262.2.16 統計方法 26第三章 實驗結果 273.1 煙麴黴分離之化合物對於 fMLF 誘發人類嗜中性白血球呼吸爆破作用和去顆粒化的影響 273.2 FM-8 抑制嗜中性白血球活化的活性並非透過細胞毒性而來 273.3 FM-8 選擇性的抑制 FPR1 刺激劑活化之嗜中性白血球細胞外超氧自由基的產生 273.4 FM-8

不具有直接清除超氧自由基的能力 283.5 FM-8 選擇性的抑制 FPR1 刺激劑活化之嗜中性白血球彈性蛋白酶的釋出 283.6 FM-8 不具抑制彈性蛋白酶活性的能力 293.7 FM-8 抑制 FPR1 刺激劑誘導下嗜中性白血球表面貼附分子 CD11b 的表現 303.8 證實 FM-8 結合至嗜中性白血球細胞膜表面的 FPR1 接受體上 303.9 FM-8 無法結合至 FPR2 受體上 313.10 FM-8 對於不同型態細胞之細胞膜表面的 FPR1 接受體也具有結合能力 313.11 證實 FM-8 透過非競爭性抑制 fMLF 刺激嗜中性白

血球的活化 323.12 FM-8 與 fMLF 活化嗜中性白血球細胞誘導的細胞內 Ca2+ 濃度變化具有拮抗性 333.13 FM-8 對嗜中性白血球 FPR1 所產生的下游訊息 Akt 與 MAPKs 之磷酸化皆有抑制 333.14 FM-8 具有還原 DPPH 的能力 343.15 FM-8 具有還原 ABTS 的能力 343.16 FM-8 具有清除過氧自由基的能力 343.17 FM-8 可以清除嗜中性白血球細胞內生成之活性氧分子 353.18 FM-8 可抑制經趨化作用之嗜中性白血球細胞的移行能力 35第四章 討論 37第五章

圖表 45參考文獻 83表目錄Table 1. The name of chemical compounds isolated from Aspergillus fumigatus. 45Table 2. Effects of pure compounds isolated from Aspergillus fumigatus on superoxide anion generation and elastase release in fMLF-induced human neutrophils. 46Table 3. Effects of FM-8 on supero

xide generation and elastase release in different stimulators-activated human neutrophils. 47Table 4. Effect of FM-8 on Ca2+ mobilization in fMLF-stimulated neutrophils. 48Table 5. Effect of FM-8 on Ca2+ mobilization induced by increasing concentrations of fMLF in human neutrophils. 49Table

6. Net AUC and ORAC values of radicals scavenging for FM-8. antioxidant activity. 50 圖目錄Figure 1. The chemical structure of FM-8. 51Figure 2. FM-8 does not cause LDH release in human neutrophils. 52Figure 3. FM-8 selectively inhibits superoxide generation in FPR1 stimulators-activated huma

n neutrophils. 54Figure 4. FM-8 does not have superoxide scavenging effect in a cell-free xanthine/xanthine oxidase system. 55Figure 5. FM-8 selectively inhibits elastase release in FPR1 stimulators-activated human neutrophils. 57Figure 6. FM-8 does not inhibit human neutrophil elastase act

ivity in cell-free systems. 58Figure 7. FM-8 selectively inhibits CD11b expression in FPR1 stimulators-activated human neutrophils. 59Figure 8. FM-8 does not inhibit CD11b expression in non-FPR1 stimulators-activated human neutrophils. 60Figure 9. FM-8 decreases FNLFNYK binding to FPR1 rece

ptor in human neutrophils. 62Figure 10. FM-8 does not decrease MMK-1F binding to FPR2 receptor in human neutrophils. 63Figure 11. FM-8 decreases FNLFNYK binding to FPR1 receptor in differentiated THP-1 cells. 64Figure 12. Effect of FM-8 on FNLFNYK binding to FPR1 receptor in FPR1-transfecte

d HEK293 cells. 65Figure 13. Non-competitive inhibitory effect of FM-8 on superoxide generation and elastase release in fMLF- stimulated human neutrophils. 66Figure 14. Non-competitive inhibitory effect of FM-8 binding to FPR1. 68Figure 15. FM-8 inhibits Ca2+ mobilization in fMLF-activatted

human neutrophils. 69Figure 16. FM-8 inhibits Ca2+ mobilization in increasing concentrations of fMLF-activated human neutrophils. 70Figure 17. FM-8 does not inhibit on Ca2+ mobilization in non-FPR1 stimulators-activated human neutrophils. 71Figure 18. FM-8 reduces phosphorylation of Akt an

d MAPKs in human neutrophils. 73Figure 19. Antioxidant effect of FM-8 in a cell-free DPPH 74Figure 20. Antioxidant effect of FM-8 in a cell-free ABTS assay. 75Figure 21. Oxygen radical absorbance capacity of FM-8 in a cell-free system. 77Figure 22. FM-8 inhibits intracellular ROS generat

ion in fMLF-stimulated neutrophils. 78Figure 23. FM-8 inhibits intracellular ROS generation in MMK-1-stimulated neutrophils. 79Figure 24. FM-8 inhibitd intracellular ROS generation in PMA-stimulated neutrophils. 80Figure 25. FM-8 inhibits fMLF and MMK-1 induced human neutrophils migration.

81Figure 26. The mechanism of FM-8 in human neutrophils. 82