臺灣博碩士論文加值系統

樟芝 (Antrodia camphorata)相傳有解毒、抗癌、解酒及消炎等多功效。本論文研究目的在探討樟芝是否同時具有抗發炎及增強免疫力之功效。分析樟芝對免疫細胞，尤其是對樹突狀細胞 (Dendritic Cells，DC)之免疫調控作用以及服食樟芝所產生之抗發炎反應。冀圖瞭解樟芝是否能經由調節促進DC的活化來達到提高免疫力？研究結果顯示，長期餵食小鼠樟芝不論對脾臟的樹突狀細胞、B細胞的數量百分比及DC上之CD40HI、CD8αHI；B細胞上之MHC class II；巨噬細胞上之CD8αHI的表現或是共同表現CD40HI、CD8αHI的樹突狀細胞群均有增加的趨勢。然DC、B細胞與巨噬細胞上CD86於服用樟芝後呈現下降的現象，以RT-PCR證實此現象與其他協同刺激分子：T細胞上CTLA-4 (Cytotoxic T lymphocyte antigen 4)、ICOS (Inducible co-stimulator)有關。亦即APC上CD86的表現下降，主要原因可能是CTLA-4參與調控平衡而抑制之。研究中發現細胞激素如INF-γ、IL-4等亦於長期服用樟芝後有上升的現象。在調查樟芝之抗發炎反應作用中，餵食小鼠樟芝後以腹腔注射LPS的方式使小鼠產生發炎反應，經流式細胞儀測定分析Trem-1表現，發現長期餵食樟芝會使Trem-1表現受到抑制而下降，證實樟芝具有抗發炎的效果。小鼠餵食樟芝四週後感染曼氏血吸蟲其免疫保護效果達到20.68%。本論文研究結果證實服食樟芝具有提升免疫力及抗發炎的功效，並了解到各種免疫反應分子間相關之作用機轉。未來更能將所使用的特異性分子作為藥物作用篩選的目標，進一步測試其他具有潛力的中草藥。

It is well known that Antrodia camphorata has many pharmacological effects, such as antidote, antineoplastic and antiphlogistic activity. In the present study, the immunomodulatroy effect of A. camphorata on dendritic cells and its anti-inflammatory action on macrophages were performed. When mice feeded with A. camphorata for 4-6 weeks, the amounts of DC and B cells; CD40HI and CD8αHI markers expressed on DCs; I-A/I-EHI molecule expressed on B cells; and CD8αHI marker expressed on macrophage increased apparently. On the other hand, the CD86 marker expressed on DCs, B cells and macrophages decreased sharply, this phenomenon may caused by the activation of CTLA-4, ICOS from T cells. The INF-γ、IL-4 mRNA cytokines, also, increased obviously after feeding with A. camphorata for 6 weeks. In addition, the expression profile of the Trem-1 protein started to decrease at 4th week and reach the lowest level at 6th week when mice administered with A. camphorata for 6 weeks, this result indicates the anti-inflammatory effect of A. camphorata. When mice administered with A. camphorata for 4 weeks and then challenged with Schistosoma mansoni, the worm reduction rate reached 20.68%. Based on these results, A. camphorata has effects on both immunomodulatory and anti-inflammation. In the future, these markers may serve as targets for futher understanding of the molecular basis and efficacy of A. camphorata and other Chinese herbs.

目錄.....................................................I
表目錄...................................................Ⅲ
圖目錄...................................................Ⅳ

中文摘要.................................................1
英文摘要.................................................2
第一章 緒論...........................................3
一、樟芝...............................................3
二、樹突狀細胞.........................................5
三、Triggering Receptors Expressed on Myeloid cell-1
(TREM-1)...........................................8
四、研究動機...........................................10
第二章 材料與方法.....................................11
一、實驗材料...........................................11
（一）樟芝萃取物來源................................11
（二）實驗動物......................................11
（三）寄生蟲來源－曼氏血吸蟲 (Schistosoma mansoni)
生活史維持....................................12
二、實驗設計...........................................13
三、實驗方法...........................................14
（一）小鼠脾臟細胞之取得............................14
（二）以流式細胞技術分析偵測小鼠脾臟免疫細胞表面
標幟..........................................15
（三）以反轉錄聚合酶連鎖反應(Reverse Transcription -
Polymerase Chain Reaction；RT-PCR)分析相關細胞
激素之mRNA表現................................17
（四）以西方墨點法測定專一性蛋白表現................20
第三章 結果...........................................24
一、樟芝對免疫細胞之調控機轉...........................24
（一）免疫細胞表面標幟測定..........................24
（二）以RT - PCR分析相關細胞激素及特殊基因之mRNA
表現..........................................32
二、樟芝抗發炎功效之細胞分子診斷.......................35
（一）Macrophage細胞表面發炎指標性Marker（Trem-1）
測定..........................................35
（二）RT - PCR分析Trem-1與發炎相關細胞激素之mRNA
表現..........................................36
（三）小鼠細胞Trem-1之蛋白表現......................36
三、樟芝提升免疫力之活體動物實驗評估...................37
第四章 討論...........................................38
參考文獻.................................................46

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