利用主動誘變與基因靜默技術提高牛樟芝菌絲體中抗發炎與抗氧化之活性成分含量

Use of activation-tagging and gene silencing technologies to enhance antioxidant and anti-inflammation functions of Antrodia cinnamomea

陳文彬(Wen-Pin Chen)

指導教授 ： 陳靖棻

朝陽科技大學/理工學院/應用化學系生化科技碩博士班/碩士(2010年)

摘要

牛樟芝，學名為Antrodia cinnamomea，為台灣特有之藥用真菌，具抗癌與降血糖之功效，也可當作抗發炎與抗氧化之用。前人研究發現經主動誘變之方式能得到一個提高牛樟芝菌絲體抗氧化與抗發炎活性成分之轉殖系菌絲體，故本試驗利用主動誘變之技術以及基因靜默(gene silencing)(antisense)兩種方式進行牛樟芝之基因轉殖，以期能獲得數個提高高抗氧化與抗發炎活性成分之菌絲體，並且找出牛樟芝菌絲體中主要之抗氧化成分與抗發炎成分。在主動誘變試驗方面成功獲得32個轉殖菌絲體(AC1～J-2)，其中AC1、AC2、AC10、AC15、AC20、AC26、AC29、J-1與J-2之抗發炎三?dehydroeburicoic acid皆顯著高於未轉殖牛樟芝菌絲體。另外與抗發炎相關的多醣β-(1-3)-D-glucan，則在AC9中較高，且粗多醣含量之多寡與β-(1-3)-D-glucan無正相關。在抗氧化分析中則顯示牛樟芝主要之抗氧化活性成分為清除自由基的酚類化合物及螯合2價離子的類黃酮。另一方面，本研究亦成功利用DMAT synthase基因靜默獲得高含量dehydrosulphurenic acid與dehydroeburicoic acid之三?之轉殖牛樟芝菌絲體AD-1。

關鍵字

牛樟芝 ； 主動誘變

並列摘要

Antrodia cinnamomea , a medicinal fungi, is well known in Taiwan as source of traditional Chinese medicine for the treatment of cancer, hyperglycemia, anti- oxidation and anti-inflammation. It has been reported that an activation-tagging method could enhance anti-inflammatory tritepenoids content in mycelia of A. cinnamomea. In order to obtain more transgenic mycelia with increased antioxidant and anti-inflammation compounds, the activation tagging method and gene silencing (antisense) of DMAT syntahse were performed separately in this study. Thirty two activation-tagged mycelia (AC1～J-2) were generated. Nine of them (AC1, AC2, AC10, AC15, AC20, AC26, AC29, J-1 and J-2) were determined with more anti-inflammatory triterpernoids (dehydroeburicoic acids), and one (AC9) with higher β-(1-3)-D-glucan (anti-inflammatory poly- saccharides). Results also showed that the phenolic compounds and flavonoids are responsible for the anti-oxidation in A. cinnamomea. By the other hand, a transgenic mycelia, AD-1, with higher dehydrosulphurenic and dehydroebucoic acids was also obtained in this study.

並列關鍵字

Antrodia cinnamomea ； activation tagging

參考文獻

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