Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/52071
標題: 薄孔菌屬萃取物之誘導肝癌細胞凋亡
Apoptosis inducing properties of extracts from Antrodia mycelia in human hepatoma cells
作者: 蔡沁玹
Tsai, Ching-Hsuan
關鍵字: Antrodia
薄孔菌屬
Antrodia salmonea
Antrodia camphorata
hepatocellular carcinoma
Sk-Hep1 cells
HepG2 cells
apoptosis
cell cycle
香杉芝
樟芝
肝細胞癌
Sk-Hep1細胞
HepG2細胞
細胞凋亡
細胞週期
出版社: 食品暨應用生物科技學系所
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摘要: 薄孔菌屬(Antrodia)是一種木材腐朽型真菌,其中香杉芝(Antrodia salmonea)為寄生於香杉木上的台灣特有真菌,於2004年被鑑定出為薄孔菌屬之新種,與台灣珍貴的藥用真菌-樟芝(Antrodia camphorata)同屬,兩者非常相似,易混淆。樟芝含有許多生理活性物質,包括:多醣體、三萜類、過氧歧化酶、腺苷及麥角固醇,在藥理活性研究中發現樟芝具有抗發炎、抗氧化、抗癌、抗腫瘤及保肝作用;近年來有學者分離出香杉芝之有效成分,發現其有抗發炎的功效以及對人類白血球細胞有抗氧化的能力(Shen et al., 2006; Shen et al., 2007)。由於香杉芝文獻甚少,因此香杉芝的生理活性及有效成分值得深入探究,本實驗之研究目的為探討香杉芝、樟芝之菌絲體萃取物誘導肝癌細胞凋亡的性質。 經分析得知香杉芝與樟芝菌絲體中含有高量的總多醣及三萜類,再分析其萃取物之總酚化合物與類黃酮之生理活性物質。 利用細胞實驗探討萃取物誘導肝癌細胞凋亡方面,將香杉芝、樟芝菌絲體之熱水和乙醇萃取物以人類肝癌細胞株(Sk-Hep1、HepG2)與人類腎臟正常上皮細胞株(293T)進行MTT試驗,結果發現香杉芝與樟芝菌絲體萃取物可依濃度升高與時間增加而提升抑制Sk-Hep1、HepG2細胞株增生的能力,香杉芝菌絲體熱水萃、乙醇萃與樟芝菌絲體熱水萃、乙醇萃對癌細胞作用72小時之生長抑制率IC50,以及在Sk-Hep1細胞株分別為2.26、1.30、2.47、1.21 mg / mL;在HepG2細胞株分別為2.26、1.30、2.11、1.55 mg / mL;另外發現萃取物對293T細胞株之生長抑制率的影響不大。 經流式細胞儀偵測其細胞週期可發現香杉芝萃取物可使Sk-Hep1細胞株停滯於G0/G1期,並可增加Sk-Hep1和HepG2細胞株之細胞凋亡的比率,但卻造成293T細胞株在高濃度1 mg / mL時大量凋亡。 當細胞走向粒線體路徑的細胞凋亡訊息傳導時,粒線體會發生膜電位下降的現象,因此使用JC-1之方法來觀察粒線體膜電位的變化,結果發現香杉芝菌絲體的熱水和乙醇萃取物對Sk-Hep1和HepG2細胞株作用皆會使粒線體膜電位有下降的趨勢。 在細胞凋亡的訊息傳導中,最後啟動了細胞內一系列的凋亡發生,引起Caspases family的活化,因此偵測Caspase-3的活性,發現香杉芝菌絲體的熱水和乙醇萃取物均可使Sk-Hep1和HepG2細胞株的Caspase-3活化,其中乙醇萃取物活化Caspase-3的能力較熱水萃取物好。 綜合上述結果可得知,香杉芝具有誘導細胞凋亡的潛力,因此推測香杉芝萃取物誘導肝癌細胞株Sk-Hep1凋亡的途徑,可由細胞週期路徑及粒線體路徑或死亡受體路徑;而HepG2細胞株可能是經由粒線體路徑或死亡受體路徑來誘導細胞凋亡,其確切影響的途徑尚須進一步深入探討。然而若要對正常細胞無傷害的情況下有效抑制癌細胞,必須降低濃度以及分離純化出指標性物質,推測可得到更佳的抑制效果,使得天然藥用菇在抗癌應用上有了新的選擇。
URI: http://hdl.handle.net/11455/52071
Appears in Collections:食品暨應用生物科技學系

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