臺灣博碩士論文加值系統

牛樟芝常做為增強免疫力、保肝、抗癌之保健食品素材，是台灣特有且珍貴的藥用真菌，近年來更為醫藥生物資源的研究重點。而本研究主要是要利用光源與培養時間差異，對液態培養牛樟芝之生長進行調控，更探討液態培養牛樟芝代謝物組成變化的影響，以及對癌細胞活性之效能。
化學分析的結果呈現，培養天數越長所得粗三萜檢測之吸光值較高，代表其代謝產物(三萜類及油酸等)的含量越高，而光源強度在15W/m2下培養之最終產物具有較高的生物活性代謝物。
抗癌活性之體外細胞實驗結果表明，液態培養牛樟芝菌絲體之乙醇萃取物對人類子宮頸癌上皮細胞(HeLa)與肝癌細胞(HuH-7)都具有效抑制性與劑量依賴關係，由培養菌絲體之醇抽出物於60μg/ml的劑量下可抑制HeLa 細胞活性達73.6%，與相同劑量下野生牛樟子實體醇抽出物的差異不大。30μg/ml的醇萃取物劑量即可抑制HuH-7 細胞活性達90%，15W/m2培養14天之菌絲體IC50劑量更僅需16μg/ml，相較於野生牛樟子實體42μg/ml是具顯著差異的。因此，本研究證明光發酵技術具有提升牛樟芝功效的應用潛力。

Antrodia cinnamomea is a unique and merit traditional medicinal fungus and used for healthy functions, such as immunity regulation, hepatoprotection, anticancer in Taiwan. The work of this study is to regulate the growth and metabolites of A. cinnamonea in submerged fermentation with color lights. And, the chemical compositions and the cyto-activities of ethanol extracts of fermentation products were evaluated by the GC/MS and the cytotoxicity assay, respectively.
The triterpenoid amounts (A245nm) and chromatograms of submerged mycelium show the complexity of chemical profiling for the increased cultured days under the illumination. The anticancer assay in vitro also indicates that the ethanol extracts have a dosage dependent inhibitory on human cervical epithelioid carcinoma (HeLa) cells, and a well-different way on human hepatoma (HuH-7) cells. The mycelium extract of 60μg/ml inhibits the 73.6% activity of HeLa cell. The 30μg/ml extract also inhibits 90% activity of HuH-7 cell. The IC50 of mycelia extract with the 15W/m2 luminance for 14 days only is 16μg/ml.
The study demonstrates the photo-fermentation has the potential to promote the functional chemicals and bio-activities of A.cinnamonea for economic production.

目錄 I
圖目錄 III
表目錄 IV
附表目錄 V
中文摘要 VI
Abstract VII
第一章研究動機與目的 1
第二章文獻回顧 5
第一節保健食品概述 5
第二節 牛樟芝 5
2.1牛樟芝簡介 5
2.2 牛樟芝之使用歷史與研究趨勢 6
2.3 牛樟芝的商業模式與生產方式 9
2.4 牛樟芝次級代謝物 11
第三章 材料與方法 20
第一節 菌株培養 20
1.1 固態培養 20
1.2 液態培養 20
1.3 液態培養基 20
1.4 固態培養基 21
第二節 儀器 21
2.1 固態與液態培養相關設備 21
2.2 分析與萃取相關設備 21
第三節 藥品 22
3.1 分析用藥品 22
3.2 萃取與分析溶劑 22
3.3 培養基 22
第四節 液態培養方法與萃取條件 22
4.1 搖瓶 22
4.2 LED光照條件設定 23
第五節機能性成分分析方法 25
5.1菌體醇抽出與粗三萜測定 25
5.2氣相層析儀條件設定 25
第六節 生物活性分析方法 26
6.1 細胞培養(Cell culture) 26
6.2細胞存活率分析(MTT assay and MTS assay) 26
第四章 結果與討論 28
第一節牛樟子實體與條件培養菌絲體之粗三萜分析 28
第二節牛樟菌層析比較分析 31
2.1 野生牛樟子實體與牛樟木氣相層析圖 31
2.2 黑暗培養組牛樟菌絲體氣相層析圖 33
2.3 15W/m2培養組牛樟菌絲體氣相層析圖 35
2.4 30W/m2培養組牛樟菌絲體氣相層析圖 37
2.5 40W/m2培養組牛樟菌絲體氣相層析圖 39
第三節 細胞存活率分析 41
3.1樟芝乙醇萃取液對HeLa cell的毒殺效果 41
3.2各培養條件之菌絲體萃取液對HeLa cell的毒殺效果 41
3.3子實體與菌絲體萃取液對HuH-7 cell的毒殺效果 45
3.4醇萃取液對HuH-7 cell型態影響 49
第四章 總結 52
參考文獻 55
附錄 62

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