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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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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。
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。
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。
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