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研究生:張莉妮
研究生(外文):Jhang, Ni-Li
論文名稱:樟芝單一化合物-去氫硫色多孔菌酸化合物 (馬偕一號)之免疫調控活性研究
論文名稱(外文):Investigation of the Immunomodulatory Activities of Dehydrosulphurenic Acid(MMH01)from Antrodia Cinnamomea.
指導教授:李金木李金木引用關係
指導教授(外文):Lee, Kin-Mu
口試委員:朱淑珍鄭柏青
口試委員(外文):Chu, Shu-ChenCheng, Po-Ching
口試日期:2014-07-24
學位類別:碩士
校院名稱:中臺科技大學
系所名稱:藥物科技研究所
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:2014
畢業學年度:102
語文別:中文
論文頁數:108
中文關鍵詞:樟芝去氫硫色多孔菌酸化合物馬偕一號免疫調控活性
外文關鍵詞:Antrodia cinnamomeadehydrosulphurenic acidMMH01immunomodulatory activities
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樟芝(Antrodia cinnamomea)亦稱牛樟芝或牛樟菇,屬於台灣獨特的菇類,只生長於台灣特有植物牛樟樹上。研究證實樟芝具有抗氧化、抗腫瘤及具免疫調節活性等特性,目前被用作為預防健康的相關傳統藥物。目前已知主要的有效成分主要為多醣體及三萜類,本研究目的在探討樟芝單一化合物-去氫硫色多孔菌酸化合物(MMH01)是否具增強免疫力之功效。
應用連續餵食不同週數與濃度的BALB/c小鼠,觀察分析餵食MMH01對小鼠脾臟抗原呈獻細胞如樹突狀細胞、巨噬細胞和B細胞與T細胞之間所產生的作用及相關細胞激素的產生,探討此樟芝MMH01之免疫調控活性及評估開發成免疫提升藥物之可行性。本研究結果顯示,樟芝- MMH01,不僅能明顯提升實驗小鼠脾臟抗原呈獻細胞數量之百分比,對於其上表面分子之表現均有明顯活化的現象如CD8αHI 、CD40HI及MHC class Ⅱ(I-A/I-E)等,顯示餵食MMH01後能夠促使DC等抗原呈獻細胞之成熟與活化。抗原呈獻細胞上表面分子CD86之表現可見隨餵食時間增長而下降。另外可見, T細胞上CTLA-4HI之表現提升,這些結果顯示抗原呈獻細胞可能與T細胞上之共同刺激分子(CD28、ICOS、CTLA-4)相互作用,而CTLA-4參與調控平衡而抑制。在細胞激素表現方面,IL-1β、TNF-α、INF-γ、IL-12的mRNA表現量顯著增加,顯示MMH01能增強實驗小鼠體內巨噬細胞分泌之細胞激素IL1β及TNF-α之表現,以及免疫細胞IL-12、IFN-γ的mRNA表現量,促使免疫走向TH1細胞免疫反應。本研究為第一個研究證實MMH01且增強免疫力之功效,本研究發現將有助於未來進一步將MMH01開發成免疫提升藥物。

Antrodia cinnamomea(AC), a unique fungus parasitic on aromatic tree Cinnamomum Kanehirai Hayata (Lauraceae) only in Taiwan. It has been used as a traditional medicine for protection of diverse health-related conditions. Recently, it has been reported that extracts and several ingredients isolated from AC possess various pharmacological and biological activities. It is well known that polysaccharides and triterpenoids are major effective components in this medicinal fungus. In this study, BALB/c mice were fed with Antrodia cinnamomea compound dehydrosulphurenic acid(MMH01) at different time points to investigate the immunomodulatory effect.
To investigate the immunomodulatory activities of the compound and evaluate the feasibility for the development of immunity-enhancing drug, the immune cells of mice such as dendritic cells(DCs), macrophages,T and B cells were analysed for the maturation and activation, the related cytokine mRNAs of the cells were also detected. When mice were administrated with Antrodia cinnamomea compound , the DC amounts and the expressions of CD40HI and CD8αHI markers on DCs increased significantly. The results indicated that the Antrodia cinnamomea compound can induce the maturation and activation of DCs. CD86 marker on DCs decreased significantly;on the other hand, the ICOSL and MHC class Ⅱ marker on macrophages increased significantly. Also, the CTLA-4HI markers on T cells increased significantly. These results indicated antigen-presenting cells interacted with the T cell costimulatory molecules of CD28, CTLA-4 and ICOS. In addition, the TNF-α, IFN-γ, IL-6 and IL-12 cytokine mRNA of mice increased significantly compared with the control group. All of the results demonstrated that AC compound possess immunomodulatory activities which lead to the immunity enhancement of the mice. This is the first report to investigate the immunomodulatory activities of a compound from Antrodia cinnamomea.

目 錄 I
表 目 錄 III
圖 目 錄 IV
中文摘要 V
Abstract VII
第一章 前言 1
一、 研究背景 1
二、 研究動機與目的 1
第二章 文獻回顧 4
一、菇類的介紹與免疫調節 4
二、樟芝介紹 5
(一)命名與分類 5
(二)分布與特徵 6
(三)生理功能 7
(四)三萜類化合物 11
三、免疫系統簡介 12
四、抗原呈獻細胞(Antigen-preseting cells;APCs) 13
五、T細胞 14
六、共同刺激分子(Costimulator Molecules) 14
七、細胞激素(Cytokines) 16
第三章 材料與方法 19
一、 實驗材料 19
(一) 去氫硫色多孔菌酸化合物之製備 19
(二) 實驗動物 19
二、 實驗設計 20
三 、實驗方法 21
(一)小鼠脾臟細胞之取得 21
(二) 以流式細胞技術分析偵測小鼠脾臟免疫細胞表面標幟 21
(三) 流式細胞技術統計分析方法 23
(四) 以即時定量反轉錄聚合酶連鎖反應(Real-Time PCR)分析相關細胞激素之mRNA表現 24
(五) Real-Time PCR反應條件設計 25
(六) Real-Time PCR統計分析方法 26
第四章 結果 27
樟芝單一化合物- MMH01對小鼠脾臟免疫細胞之調控 27
(一) 免疫細胞表面標記測定 27
1.小鼠脾臟細胞-樹突狀細胞(DCs)之表現 27
2.小鼠脾臟細胞-巨噬細胞(Macrophage)之表現 29
3.小鼠脾臟細胞-B細胞之表現 32
4.小鼠脾臟細胞-T細胞之表現 34
(二)以Real-time PCR分析相關細胞激素之mRNA表現 38
(1)IL-1β mRNA之表現 38
(2)IL-4 mRNA之表現 39
(3)IL-6 mRNA之表現 39
(4)IL-10 mRNA之表現 40
(5)IL-12 mRNA之表現 40
(6)IL-18 mRNA之表現 41
(7)INF-γ mRNA之表現 41
(8)TNF-α mRNA之表現 42
(9)iNOS mRNA之表現 42
第五章 討論 43
第六章 結論 53
參考文獻 54

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