臺灣博碩士論文加值系統

牛樟芝(Antrodia cinnamomea)是台灣特有藥用菇菌，僅生長於台灣特有之牛樟
樹(Cinnamomum kanehirai) 的腐朽心材内壁，或枯死倒伏之牛樟樹幹的潮濕表面，
近來出現變種白色牛樟芝，同為臺灣著名且珍貴之藥用真菌，近來科學研究發現
其可能具有醫療功效。
目前對於白樟芝的研究甚少，因此本研究從事固態培養紅樟芝與白樟芝，比
較酚類及粗三萜生物活性成分、抗氧化能力與對前列腺癌細胞之抗轉移詴驗、與
抗增生詴驗，並以流式細胞儀PI 染色法觀察細胞週期分佈，探討抗前列腺癌之作
用機制。
結果顯示紅樟芝與白樟芝熱水萃取物酚類含量最高，乙醇萃取物粗三萜含量
最高。紅樟芝抗氧化能力優於白樟芝。紅樟芝與白樟芝熱水萃取物皆能夠給予
CHO-K1 細胞增加抗氧化能力，兩者保護能力明顯優於修復能力。樟芝抗前列腺癌
細胞DU-145 活性，紅樟芝與白樟芝乙酸乙酯萃取物抑制增生能力最佳，白樟芝抑
制轉移能力優於紅樟芝。以紅樟芝與白樟芝乙酸乙酯萃取物處理前列腺癌細胞
DU-145，可使DU-145 細胞週期停滯在G2/M 期，抑制癌細胞增生。

Antrodia cinnamomea is well known in Taiwan as an expensive medicinal fugi.
Recently,. Both red and white Antrodia cinnamomea (RAC and WAC) are folk
medicinal mushroom commonly used in Taiwan, the latter recognized as a variant of
Antrodia cinnamomea.
In this study, we respectively used solid substrates to culture RAC and WAC strain
by solid-state fermentation, compared different extracts to evaluated phenolic content,
crude triterpenoids content, antioxidant capacity and antiproliferatiive effect on DU-145
prostate cancer cells.
The results showed that the phenolic content of hot water (HW) extract and the
crude triterpenoids content 95% ethanol extract (EtOH) own the highest conetnt. HW
extract can provide CHO-K1 cells to resist the oxidative stress from H2O2. The
protective antioxidant capacity of RAC HW extract is better than that of WAC. RAC
and WAC ethyl acetate (EA) own the best antiproliferation in DU-145 cells, and WAC
inhibited DU-145 migration ability was significantly better than RAC. RAC and WAC
EA extracts treatment DU-145 cells through G2/M phase cell cycle arrest and induction
of apotosis.in. Our result showed that RAC and WAC own antioxidant capacity, and anti
prostate cancer, suggesting that it may be a potential preventive or therapeutic prostate
cancer.

摘要 ...................................................................................................................................I
ABSTRACT..................................................................................................................... II
目次 ................................................................................................................................ III
表目錄 ............................................................................................................................VI
圖目錄 ...........................................................................................................................VII
第一章、緒論 .................................................................................................................. 1
1.1 前言...................................................................................................................... 1
第二章、文獻回顧 .......................................................................................................... 2
2.1 抗氧化概述............................................................................................................ 2
2.1.1 自由基對生物體的影響................................................................................ 2
2.1.2 抗氧化劑........................................................................................................ 4
2.2 前列腺癌................................................................................................................ 8
2.2.1 前列腺和前列腺癌的介紹............................................................................. 8
2.2.2 前列腺癌的轉移與侵略................................................................................. 8
2.2.3 前列腺癌的分類與分期................................................................................. 9
2.2.4 前列腺癌的治療........................................................................................... 10
2.3 牛樟芝簡介.......................................................................................................... 11
2.3.1 牛樟芝命名及分類....................................................................................... 11
2.3.2 牛樟芝的形態............................................................................................... 13
2.3.3 牛樟芝的活性成份....................................................................................... 14
2.3.4 牛樟芝的藥理活性....................................................................................... 16
2.4 固態與液態發酵.................................................................................................. 20
2.5 研究動機.............................................................................................................. 23
2.6 研究架構.............................................................................................................. 24
第三章、材料與方法 .................................................................................................... 25
3.1 實驗材料.............................................................................................................. 25
3.1.1 設備............................................................................................................... 25
3.1.2 藥品............................................................................................................... 26
3.2 牛樟芝固態發酵產物製備及細胞培養 .............................................................. 28
3.3 分析方法 .............................................................................................................. 31
3.3.1 Folin-Ciocalteu’s phenol reagent ................................................................... 31
3.3.2 Vanillin-Acetic acid 香草醛-冰醋酸法........................................................ 32
3.3.3 ABTS 自由基清除能力............................................................................... 33
3.3.4 OH 自由基清除能力..................................................................................... 35
3.3.5 還原力........................................................................................................... 36
3.3.6 比較不同萃取物對固態發酵酚含量及抗氧化能力................................... 37
3.3.7 紅樟芝與白樟芝熱水萃取物對CHO-K1 細胞抗氧化能力....................... 37
3.3.8 比較不同萃取物對固態發酵粗三萜含量及細胞毒性測詴-MTT assay.... 38
3.3.9 細胞形態觀察(cell morphology) ................................................................ 39
3.3.10 轉移詴驗(migration assay)........................................................................ 39
3.3.11 細胞週期測定(Cell Cycle Analysis)-Propidium (PI) stain ......................... 40
4.1 固態發酵不同萃取物對酚類含量、粗三萜含量及抗氧化能力之比較.......... 41
4.2 紅樟芝與白樟芝熱水萃取物對 CHO-K1 細胞抗氧化能力.............................. 44
4.3 比較固態發酵不同萃取物對細胞毒性測詴-MTT ASSAY................................. 48
4.4 紅樟芝與白樟芝乙酸乙酯萃取物處理 DU-145 細胞之形態變化................... 55
4.5 比較紅樟芝與白樟芝乙酸乙酯萃取物抑制DU-145 細胞之轉移能力........... 57
4.6 紅樟芝與白樟芝乙酸乙酯萃取物對 DU-145 細胞之細胞週期的影響........... 60
第五章、結論................................................................................................................ 65
第六章、參考文獻 ........................................................................................................ 66

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