臺灣博碩士論文加值系統

新日本靈芝是一種罕見寄生在腐敗竹子的多孔菌，然而此靈芝在抑制過敏反應的效用還未被探討，本研究以Anti-DNP IgE/DNP-BSA抗原或鈣離子載體A23187誘導RBL-2H3肥大細胞的模式來測試新日本靈芝水萃物(GNES)的抗過敏效應，結果顯示，GNES可以抑制組織胺和β- hexosaminidase的分泌，並減少介白素-4和腫瘤壞死因子-α等發炎性細胞素生成，此外，GNES也會降低肥大細胞白三烯素的產生。為進一步研究GNES抑制肥大細胞脫顆粒，降低發炎性細胞素和白三烯素生成機制，我們分析肥大細胞蛋白激酶C-μ（PKC-μ）、絲裂原活化蛋白激酶（mitogen-activated protein kinases, MAPKs) 及細胞質磷脂質A2 (cytosolic phospholipases A2, c-PLA2)的活化情形，實驗結果顯示，GNES可抑制Anti-DNP IgE/DNP-BSA或A23187誘導的PKC-μ，c-PLA2及ERK蛋白的磷酸化，進而抑制肥大細胞脫顆粒、脂質調節物及促發炎激素的生成。本結果顯示應用新日本靈芝水萃物在第一型過敏治療上是具有潛力的。

Mast cells play an important role in allergic responses because they produce histamine, lipid mediators, such as leukotrienes (LTs), prostaglandins (PGs), and a variety of inflammatory cytokines. Thus, pharmacological interventions for allergies have focused on mast cell inactivation. Ganoderma neo-japonicum (GN) is a rare polypore mushroom found growing on decaying bamboo; however, its inhibitory effects on allergic reactions have not been determined. In this study, water extract of Ganoderma neo-japonicum (GNES) was used to test its anti-allergy effect in Anti-DNP IgE/DNP-BSA or Ca2+ ionophore A23187 stimulated rat RBL-2H3 mast cells. The results showed that GNES could inhibit secretion of histamine and beta-hexosaminidase and reduced the production of pro-inflammatory cytokines, such as interleukin-4 and tumor necrosis factor-α. GNES also decreased the production of leukotriene C4. To investigate the underlying mechanisms of GNES on degranulation, cytokine and leukotriene production, we examined the activation of protein kinase c (PKC) and mitogen-activated protein kinases (MAPKs). The data showed that GNES suppressed Anti-DNP IgE/DNP-BSA or Ca2+ ionophore A23187 induced phosphorylation of protein kinase c (PKC) μ, cytosolic phospholipases A2 (c-PLA2), and extracellular signal–regulated kinases (ERK). These results indicated that GNES may be a useful therapeutic agent for immediate-type hypersensitivity.

致謝 --------------------------------------------------------------------- i
中文摘要 --------------------------------------------------------------------- iii

英文摘要 --------------------------------------------------------------------- iv
目錄 --------------------------------------------------------------------- v
圖目錄 --------------------------------------------------------------------- vii

名詞縮寫表 --------------------------------------------------------------------- ix
第一章 文獻回顧
第一節 過敏--------------------------------------------------------------- 1
第二節 新日本靈芝------------------------------------------------------ 2
第三節 肥大細胞活化指標及調控------------------------------------ 5
第四節 臨床治療過敏之藥物副作用--------------------------------- 9
第五節 研究動機與目的------------------------------------------------ 11
第二章 材料與方法
第一節 研究材料--------------------------------------------------------- 12
一 新日本靈芝水萃物
二 細胞培養
三 實驗藥物
四 細胞存活率分析
五 β-hexosaminidase分析
六 Histamine分析
七 TNF-α偵測
八 Leukotriene C4偵測
九 反轉錄聚合酶鏈反應
十 西方墨點法分析
十一 儀器相關設備
第二節 研究方法--------------------------------------------------------- 22
一 新日本靈芝水萃液製備

二 細胞培養
三 Anti-DNP IgE/DNP-BSA及A23187處理方式
四 細胞存活率分析
五 β-hexosaminidase活性分析
六 Histamine分析
七 TNF-α偵測
八 Leukotriene C4偵測
九 反轉錄聚合酶鏈反應
十 西方墨點法分析
十一 統計分析
第三章 研究結果--------------------------------------------------------- 31
第四章 結果與討論------------------------------------------------------ 43
參考文獻 --------------------------------------------------------------------- 45
附錄 ---------------------------------------------------------------------
附錄一 --------------------------------------------------------------------- 66

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