桑葚抗氧化分析及對肝癌細胞毒性研究

Study of antioxidative activities and cytotoxicity on liver cancer cells of mulberry

王麗娟(Li-Chuan Wang)

指導教授 ： 周芬碧

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2010年)

https://doi.org/10.6834/CSMU.2010.00031

摘要

過去研究對於桑葚(Mulberry water extracts, ME)的多種成分分析、抗氧化程度和抗癌能力尚未明確，因此本研究首先對於桑葚醣類、蛋白質、脂肪、多酚、類黃酮和膳食纖維的成分進行分析。另一方面，桑葚含有花青素，而花青素具有抗氧化能力，所以測試DPPH和總抗氧化能力分析(TRAP)；在DPPH實驗結果，ME在濃度在2.5 mg/ml時，自由基清除率可以達50%，在總抗氧化TRAP分析結果可看出100μg/ml的ME具有與0.158μM trolox有相同的抗氧化能力，因此推測桑葚確實具有抗氧化的能力。接著各挑選2株常見的人類大腸癌細胞(CoLo-205, CaCo-2)和2株常見的肝癌細胞(Hep 3B、Hep G2)，進行細胞毒殺測試。實驗結果發現ME對於肝癌細胞的毒殺效果較佳，進一步利用DAPI以及AVO螢光染色分析，極可能是走向細胞凋亡的途徑。另外，現今癌症治療多以放射性照射或合併化學藥物的方式進行，然而這類的治療常會造成患者的不適與許多後遺症，因此將桑葚萃取物合併放化療之後是否能達到降低劑量以及更佳的治療效果，實驗結果證實桑葚輔以放射線和paraplatin的抗癌藥物作用下，Hep 3B和Hep G2細胞並沒有加成毒殺的效果。

關鍵字

桑葚 ； 抗氧化力 ； 細胞凋亡

並列摘要

In the past, there is no clear definition for ME’s ingredients analysis, antioxidative activities, and cytotoxicity on cancer cells. Therefore, we analyze total poly-carbohydrates, phenolics, favonoids , protein and fat at mulberry water extracts. In the other hand, Mulberry fruit is rich in anthocyanins which have been reported to be beneficial to health as potent antioxidants, so we tested antioxidative ability of mulberry water extract by DPPH and TRAP assay. We demonstrated that the extract of mulberry induced a strong cytotoxic effect toward Hep 3B and HepG2 cells. The mechanisms of the cytotoxic effect in mulberry water extracts were concentration-dependent. High doses of ME induced apoptotic cell death in Hep 3B and Hep G2 cells. Therefore we suggest that it may provide a method to treat liver cancer.

並列關鍵字

mulberry ； antioxidative ability ； apoptosis

參考文獻

陳建仁(2004)。中華中醫藥典。行政院衛生署，124。

Alexandrakis, M., Letourneau, R., Kempuraj, D.,Kandere-Grzybowska, K., Huang, M., Christodoulou,. S., Boucher, W., Seretakis, D., and Theoharides, T.C. (2003). Flavones inhibit proliferation and increase mediator content in human leukemic mast cells (HMC-1). European journal of haematology. 71: 448-454.

Alho H.,Leinonen J.(1999). Total antioxidant activity measured by chemiluminescence methods. Methods Enzymol, 2999: 3-15

Alva AS, Gultekin SH, Baehrecke EH. (2004). Autophagy in human tumors:cell survival or death? Cell Death Differ 11(9): 1046-1048.

Asano N, Yamashita T, Yasuda K et al. (2001).Polyhydroxylated alkaloids isolated from mulberry trees (Morusalba L.) and silkworms (Bombyx mori L.). J Agric Food Chem. 9:4208-13.