碩士 / 國立臺灣海洋大學 / 食品科學系 / 105 / Marine toxins are widely found in many marine organisms with species diversity. Palytoxin (PTX) is a non-protein marine toxin, which could cause food poisoning by food chain accumulation. PTX is found in Palythoa spp. and Ostreopsis spp. The frequent symptoms of PTX poisoning are including fever, nausea, vomit, breathing difficulties, arrhythmias and muscle pains caused by rhabdomyolysis. Taiwan, an area in the substropical, is surrounded by the sea and PTX producers exist in Taiwan waters. In the past, food poisoning events happened as eating the marine organisms. Hence, we should know about PTX toxicity.
In the first chapter, DNA sequence of zoanthids collected from the coastal of Da-Ping, Keelung, Taiwan were identified. By using polymerase chain reaction (PCR), mitochondrial DNA (mtDNA) was chosen as the target DNA, then we set cytochrome c oxidase I (COI) and 16S rRNA as a target gene in this studt. Finally the DNA sequences of zoanthids were compared with the data base in National Center for Biotechnology Information (NCBI), and the zoanthids are identified as Zoanthus kuroshio, Palythoa mutuki, Palythoa caribaeorum and Palythoa tuberculosa. For the detection method of toxin, we use mouse neuroblastoma cell line (Neuro-2a) as cell model. Ouabain, the competitive inhibitors targeted on Na+/K+-ATPase was added. Four bioactivity indicators contain cell viability, lactate dehydrogenase (LDH), creatine kinase (CK) and adenosine triphosphatase (ATPase). The results indicate cells pre-incubated with ouabain can increase sensitivity and the bioactivity indicators have dose-dependent.
In the second chapter, Neuro-2a cells were treated with PTX (1.56~100 pM) for 24 hr to investigate the toxicological effects. In the pathways, toxins could induce reactive oxygen species (ROS) and nitric oxide (NO) increase and cause oxidative stress. Meanwhile, mitochondrial membrane potential (MMP) and production of adenosine triphosphate (ATP) decrease, which have dose-dependent finally activate the cysteine dependent aspartate-specific protease (caspase). On the other hand, using Western blot to analyze endoplasmic reticulum stress performance find amount of performance of related regulatory protein glucose-regulated protein 78 (GRP78) is dramatically increased. The apoptotic related protein Bak and Bcl-2, which ratio (Bak/Bcl-2) have significant increase in the study. The results suggest that Neuro-2a cells were induced to mitochondrial-related apoptosis after exposed to different doses of PTX (1.56~100 pM).
| Identifer | oai:union.ndltd.org:TW/105NTOU5253006 |
| Date | January 2016 |
| Creators | Wu, Shang-Yi, 吳尚宜 |
| Contributors | Hwang, Deng-Fwu, 黃登福 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 84 |
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