牛樟芝（Antrodia cinnamomea, AC）是一種台灣特有的藥用真菌，AC的生理活性包含護肝、抗發炎、抗C型肝炎病毒及抗癌等；因此，AC已成為新藥開發及保健食品研發的重要原料。中草藥及藥物奈米化已被證實可增加溶解度、生物利用率、減少藥物毒性、強化藥理活性等功能；但目前很少有奈米化AC之活性物質含量及生理活性的文獻報導。本實驗嘗試比較一般研磨牛樟芝子實體（Ground Antrodia cinnamomea fruiting body, GACFB）及奈米研磨牛樟芝子實體（Nanonized Antrodia cinnamomea fruiting body, NACFB）其三萜類、多醣體含量及以LPS誘導發炎的RAW264.7細胞之抗發炎活性之差異。結果顯示NACFB 在UPLC-MS/MS分析之三萜類含量antcin K、antcin H、 antcin B、antcin C、dehydroeburicoic acid等5種三萜類的含量分別比GACFB高出 67%、 66%、 54%、 53% 及 51%。NACFB多醣體含量較GACFB高18.51%，並具有顯著差異。在RAW264.7細胞抗發炎活性中NACFB在6.25 μg/mL下對MAPKs訊號之磷酸化JNK、ERK1/2、p38表現量抑制效果顯著高於GACFB 13.8%、24.8%及34.8%。在PGE2表現量與細胞核之轉錄因子p65表現量顯著高於GACFB 10.8%、 28.3%。在IL-6、IL-10抑制效果顯著高於GACFB 45.84 %、81.27%，結果顯示在相同的濃度下NACFB比GACFB更能有效的抑制由LPS誘導RAW264.7細胞的發炎反應。

Antrodia cinnamomea (AC), a Taiwan-specific Medicinal mushroom, has been reported to have numerous biological activities including hepatoprotection, anti-inflammation, antihepatitis C virus activity, and anticancer activity. It has been used in the formulation of pharmaceutical and functional foods. Nanotechnology has large number of advantages for herb or drugs, including enhancement of solubility and bioavailability, protection from toxicity, enhancement of pharmacological activity, enhancement of stability, improving tissue macrophages distribution, sustained delivery, protection from physical and chemical degradation. However, very few researches have been done to investigate the effects of nanoization on the chemical composition and biological functions of AC. The amount of crude polysaccharides, triterpenoids and anti-inflammatory activity of Ground Antrodia cinnamomea fruiting body (GACFB) and Nanonged Antrodia cinnamomea fruiting body (NACFB) in RAW264.7 macrophages treated with LPS were compared in this study. The results from UPLC-MS/MS analyses showed the amount of triterpenoids (antcin K, antcin H, antcin B, antcin C, dehydroeburicoic acid) in NACFB were increased 67%, 66%, 54%, 53% and 51% respectively when comparied with GACFB. The amount of polysaccharide increased 18.51% after nanoization. The inhibitory effects of NACFB on phosphor-JNK, ERK1/2, p38, p65, PGE2, IL-6 and IL-10 in RAW264.7 cells was significantly higher than that of GACFB (13.25, 24.8, 34.8, 28.3, 10.8, 45.84 and 81.27%) at the concentration of 6.25 μg/mL. The results from this study indicate the anti-inflammatory activity of Antrodia cinnamomea fruiting body was increased significantly after nanoization.

摘要 I
Abstract III
謝誌 IV
目錄 V
表目錄 VII
圖目錄 VIII
縮寫表 X
第一章、 前言 1
第二章、 文獻探討 3
一、 發炎反應 3
1. 發炎機轉 4
2. 脂多醣 ( Lipopolysaccharide, LPS ) 與發炎反應 12
3. 炎症細胞 ( Inflammatory cells ) 14
4. 炎性細胞激素 ( Inflammatory cytokines ) 16
5. 其他與發炎相關蛋白質或賀爾蒙 ( Other inflammatory-related proteins or hormones ) 18
二、 牛樟芝 ( Antrodia cinnamomea, AC ) 20
1. 三萜類 ( Triterpenoids ) 20
2. 多醣體 ( Polysaccharides ) 24
三、 奈米顆粒 ( Nanoparticles ) 25
四、 研究目的 29
五、 研究假設 30
第三章、 材料與方法 31
一、 實驗架構 31
二、 試劑與材料 32
三、 藥品 32
四、 儀器 33
五、 實驗方法 35
2. UPLC/MS/MS三萜類定性、定量 36
3. 粗多醣含量測試 37
4. 小鼠 RAW264.7 cells 培養 38
5. 細胞存活率測定 39
6. 蛋白質表現分析 39
7. 酵素連結免疫吸附分析 42
8. 統計分析 44
第四章、 結果 45
一、 GACFB粉末光學顯微鏡相片 45
二、 NACFB在電子顯微鏡粒徑分析 45
三、 GACFB及NACFB在UPLC/MS/MS 圖譜及定性定量結果 45
四、 GACFB與NACFB粗多醣測試結果 46
五、 GACFB與NACFB對RAW264.7細胞之毒性測試 46
六、 GACFB與NACFB對經LPS誘導之RAW264.7細胞之磷酸化JNK、ERK1/2、p38 蛋白質表現量之影響 46
七、 GACFB與NACFB對經LPS誘導之RAW264.7細胞之COX-2、PGE2蛋白質表現量之影響 47
八、 GACFB與NACFB對經LPS誘導之RAW264.7細胞之細胞核轉錄因子p65蛋白質表現量之影響 48
九、 GACFB與NACFB對經LPS誘導之RAW264.7細胞之促發炎激素IL-1β之影響 48
十、 GACFB與NACFB對經LPS誘導之RAW264.7細胞之促發炎激素IL-6之影響 49
十一、 GACFB與NACFB對經LPS誘導之RAW264.7細胞之促發炎激素IL-10之影響 49
十二、 GACFB與NACFB對經LPS誘導之RAW264.7細胞之促發炎激素IL-18之影響 50
第五章、 討論 51
第六章、 結論 55
第七章、 參考文獻 57

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