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1、香椿萃取物成分鑑定及誘導人類肝癌細胞凋亡之探討Identification of the compounds of Toona sinensis extracts and induction of apoptosis in human hepatoma (HepG2) cells研究生：江靜宜 Jiang, Jing-Yi 指導教授：黃卓治 Huang, Tzou-Chi 【摘要摘要】香椿 (Toona sinensis) 屬楝科植物，廣佈於亞洲各地，在中國社會中已長時間被當作營養食品用。gallic acid (GA)、methyl gallate (MG)及 1,2,3,4,6-penta
2、-O-galloyl-D-glucose (5GG) 是香椿萃取物中的主要成份，三者皆具有沒食子酸的基本結構。本實驗利用 50%甲醇萃取香椿葉後，並以液相層析儀 (HPLC)、液相層析質譜儀 (LC-MS) 及核磁共振質譜儀 (NMR) 分析鑑定其化學結構。結果顯示，香椿萃取物中 gallic acid 含量則是 0.069 mg/100 mL，methyl gallate 為 1.184 g/100 mL，5GG 為 1.36 g/100 mL。此外，進一步以細胞模式實驗探討香椿萃取物調控人類肝癌細胞HepG2 凋亡之能力。實驗證實，香椿萃取物可有效抑制人類肝癌細胞 HepG2 的增生並誘導
3、細胞凋亡。其誘導細胞死亡的特徵包括細胞型態的改變、核染色質凝集及 DNA 片斷化。實驗中使用流式細胞儀測量 Sub-G1 特徵，結果顯示香椿萃取物會使 HepG2 依 concentration-及 time-dependent的趨勢而停止生長，且發現香椿萃取物對於 HepG2 細胞會有 S/G2 細胞週期停止的現象，並藉由造成細胞粒線體膜電位、reactive oxygen species (ROS)及 glutathione (GSH) 的改變而導致細胞凋亡。關鍵字：香椿萃取物、液相層析儀、液相層析質譜儀、凋亡【Abstract】Toona sinensis Roem. (Meliacea
4、e; T. sinensis), a kind of arbor, is widely distributed in Asia and long used as a nutritious food in Chinese society. gallic acid (GA)、methyl gallate (MG) and 1,2,3,4,6-penta-O-galloyl-D-glucose (5GG) are the major compounds of the Toona sinensis extracts. All of them contain galloyl group as the m
5、ajor part. In this study, we extracted Toona sinensis young leaves by 50% methanol. High performance liquid chromatography (HPLC), high performance liquid chromatography mass (LC-MS) and nuclear magnetic resonance spectroscopy (NMR) were used to elucidate chemical structure. The result showed that T
6、oona sinensis contains gallic acid 0.069 mg/100 mL, methyl gallat 1.184 g/100 mL, 5GG 1.36 g/100 mL. Cell model system was set up to modulate the apoptotic effect of Toona sinensis extracts. This data showed that Toona sinensis extracts was effective as an anti-proliferative agent of human cancer ce
7、lls and induction of apoptosis in HepG2 cells. We used flow cytometry to determine the Sub-G1. The result showed that Toona sinensis extracts was able to induce cell arrests in concentration- and time-dependent manner. We also found that Toona sinensis extracts treatmemt could induce S/G2 phase cell cycle arrest and induction of apoptosis in HepG2 cells through modulation of mitochondrial functions, reactive oxygen species (ROS), andglutathione (GSH).Key words: Toona sinensis extracts, HPLC, LC-MS, apoptosis
