碩士 / 嘉南藥理大學 / 化粧品應用與管理系 / 103 / Part1
The bioactivity-guided fractionation isolation the extract of traditional herbs of RCE, COE, LLE and LPE in ethanol, n-hexane, ethyl acetate and n-butanol phase was investigated the anti-oxidation, anti-melanogenesis, and protection of plasmid DNA. We selected the extracts for improving skin cell proliferation, and then the protection and repair of skin cells of the extracts were examined. The extract of RCE in n-butanol phase expressed 91.7% DPPH free radical scavenging activity and rich polyphenols and flavonoids. The extract of RCE in ethyl acetate phase inhibited 27.6% NO free radical activities. The extract of COE in ethyl acetate phase had great effect of ABTS free radical scavenging activity (83.7%) and suppressed 98% lipid peroxidation production. Furthermore, LLE decreased intracellular ROS (74.8%) and increased GSH (32.1%) production in H2O2-treated human keratinocyte HaCaT cells. The extract of LLE in n-hexane phase could significantly suppress the cellular tyrosinase activity (92%), DOPA oxidase activity (100%) and melanin content (69%). Additionally, we also found that RCE evidently had a greater effect to keep DNA integrity and protect skin cells against UVB. RCE obviously decreased erythema formation after UVB-induced acute inflammation in male SKH-1 hairless mice. Therefore, we suggested that RCE has ability to prevent inflammation reaction from UVB irradiation. Base on all above these experiments, we concluded that RCE has strong potential to be developed.
Part2
In this study, we investigated the mechanism of anti-skin cancer though two pure compounds (1) and (2), which from marine lives. The cell viability in human epithelial carcinoma cell (A431) and human squamous carcinoma cell (SCC25) had been demonstrated by MTT assay. We found that treatment with (1) and (2) had apparently cytotoxicity and dose-dependent effects on A431 and SCC25 cells after 72 h. Morphological observation on cell death induced by (1) and (2) in A431 and SCC25 cells. Following treatment with (1) and (2) in both cancer cells, we found the typical features such as cell shrink, dynamic membrane blebbing and chromatin condensation. Analyzing the periodically changed of cell cycle, we showed that (1) and (2) led both cells arrest in G0/G1 phase with a concomitant significantly increased of sub-G1 population. In the intracellular reactive oxygen species (ROS) and glutathione (GSH) experiments, we found both compounds reduced the GSH and increased ROS generation. Further, we investigated the expression of apoptosis-associated factors by immunofluorescence and western blotting, results showed that (1) and (2) upregulated the expression of p53, p21, bax, cytochrome c, Fas, FasL, TRAIL, TRAILR1, TRADD, FADD, Caspase-8, Caspase-9 and Caspase-3. Therefore we deducted that (1) and (2) possibly induced apoptosis through bath of mitochondrion and death receptor pathway. These observations show that (1) and (2) have an anti-cancer effect which may be related with the prevention of cell proliferation, and promotion the activities of oxidative stress and cell apoptosis-related proteins in the skin cancer cells.
| Identifer | oai:union.ndltd.org:TW/103CNUP0115021 |
| Date | January 2015 |
| Creators | Chiou-Rung Wu, 吳秋瑢 |
| Contributors | Chia-Hua Liang, 梁家華 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 128 |
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