碩士 / 國立高雄海洋科技大學 / 水產食品科學研究所 / 94 / The apoptosis-induced fractionation and purification was used to identify the bioactive components of Hard clams (HC), Meretrix lusoria. The effects of ethyl acetate extract of HC (HC-EA) on cell viability in human cancer cells were investigated. Dissipation of mitochondrial membrane potential, release of mitochondrial cytochrome c into cytosol and subsequent induction of pro-caspase-9 and -3 processing preceded confirmation of induction of apoptosis in HL-60 cells by DNA fragmentation, chromatin condensation, changes in the cell membrane and the appearance of a sub-G1 DNA peak. Furthermore, treatment with HC-EA caused a rapid loss of intracellular glutathione content and stimulation of reactive oxygen species (ROS). Antioxidants such as catalase, N-acetylcysteine, pyrrolidine dithiocarbamate, and superoxide dismutase, but not allopurinol and diphenylene iodonium, significantly inhibited HC-EA-induced cell death. Apoptosis was completely prevented by a pan-caspase inhibitor, z-Val-Ala-Asp-fluoromethyl ketone (z-VAD-FMK). The ethyl acetate fraction was then subjected to separation and purification using silica gel column chromatography. The compounds showing strong apoptosis inducing activity were identified by spectral methods as: 5α,8α-epidioxy-24(S)-methylcholest-6-en-3β-ol and 5α,8α- epidioxy-24(R) -methylcholest-6-en-3β-ol.
| Identifer | oai:union.ndltd.org:TW/094NKIMT084004 |
| Date | January 2006 |
| Creators | Yu-Ting Huang, 黃郁婷 |
| Contributors | Min-Hsiung Pan, 潘敏雄 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 138 |
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