碩士 / 國立臺灣海洋大學 / 食品科學系 / 97 / Abstract
In the present study, Sargassum and Spirulina polysaccharides extracts were degraded by Pseudomonas vesicularis MA103 and Aeromonas salmonicida MAEF108 agarases to yield Sargassum and Spirulina oligosaccharides hydrolysate fraction of < 1 kDa, 1-3 kDa, and 3-5 kDa to perform anti-oxidative activity assays. And the better anti-oxidative algal samples are following evaluated whether treatment with Sargassum or Spirulina oligosaccharides hydrolysate fraction reduced arsenic-induced oxidative stress in PC12 neuronal cells. Experimental data showed that 3-5 kDa Sargassum oligosaccharides hydrolysate fraction contained the highest total sugar content and reducing sugar content (870.94 ± 67.16 �慊/mL and 0.62 ± 0.01 mg/mL), and 3-5 kDa Spirulina oligosaccharides hydrolysate fraction contained the highest sulfated content (32.20 ± 0.11%). In terms of anti-oxidative activities, the total phenolic content was highest in the 3-5 kDa Sargassum oligosaccharides hydrolysate fraction (86.67 ± 0.88 �慊/mL). In evaluating the scavenging effects on DPPH, Sargassum oligosaccharides hydrolysate fraction performed better, while < 1 kDa Spirulina oligo- saccharides hydrolysate fraction had better chelating effects on ferrous ions. In vitro cell studies showed that As3+-treated PC12 neuronal cells had significantly reduced cell viability, while those cotreated with 1-3 kDa Sargassum, < 1 kDa, 1-3 kDa, or 3-5 kDa Spirulina oligosaccharides hydrolysate fraction had significantly increased cell viability relative to As3+ only group, with increases of 7.2%, 11.9%, 7.4%, and 13.6% cell viability, respectively. Additionally, when given 1-3 kDa Sargassum, < 1 kDa, 1-3 kDa, or 3-5 kDa Spirulina oligosaccharides hydrolysate fraction with As3+, reactive oxygen species (ROS) production of PC12 neuronal cells was inhibited, with decreases of 55%, 61%, 58%, and 52%, respectively, relative to As3+ group. Furthermone, intracellular glutathione (GSH) level changed after treating PC12 neuronal cells with As3+ and results showed that when 1-3 kDa Sargassum, < 1 kDa, 1-3 kDa, or 3-5 kDa Spirulina oligosaccharides hydrolysate fraction was given with As3+, intracellular GSH content of PC12 neuronal cells increased by 98%, 89%, 87%, and 115%, respectively, relative to As3+ group. Using western blotting to analyze heat shock protein 70 (HSP 70) and heme oxygenase-1 (HO-1) expression, results indicated that 1-3 kDa Sargassum oligosaccharides hydrolysate fraction significantly inhibited As3+-induced HSP 70 and HO-1 expression by 13% and 14% when compared to As3+ group, and 3-5 kDa oligosaccharides hydrolysate fraction inhibited by 50% and 48%. After treatment with 50 �嵱 As3+ for 24 hr, intracellular phosphorylated extracellular signal-regulated kinases (ERK) expression of PC12 neuronal cells significantly increased. However, regardless of whether Sargassum or Spirulina oligosaccharides hydrolysate fraction was added, phosphorylated ERK expression was not significantly reduced. In terms of changes in phosphorylated c-Jun N-terminal kinases (JNK) expression, giving As3+ with 1-3 kDa Sargassum, 1-3 kDa, or 3-5 kDa Spirulina oligosaccharides hydrolysate fraction significantly decreased phosphorylated JNK expression (from 33% to 46%). Lastly, after treatment of 50 �嵱 As3+ with 1-3 kDa Sargassum, < 1 kDa, 1-3 kDa, or 3-5 kDa Spirulina oligosaccharides hydrolysate fraction for 24 hr, phosphorylated P38 expression was noticeably reduced in PC12 neuronal cells (from 28% to 39%). In conclusion, Sargassum and Spirulina oligosaccharides hydrolysate fraction reduce As3+-induced oxidative stress and cell death in PC12 neuronal cells, serving as the basis for development of anti-oxidative agents in the future, thus creating a novel form of health food products.
| Identifer | oai:union.ndltd.org:TW/097NTOU5253071 |
| Date | January 2009 |
| Creators | Su-Fen Fan, 范素芬 |
| Contributors | Chorng-Liang Pan, Anya Maan-Yuh Lin, 潘崇良, 林滿玉 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 103 |
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