碩士 / 長庚大學 / 基礎醫學研究所 / 89 / A large outbreak of enterovirus 71 occurred in Taiwan in 1998. More than hundred thousands of people got infected, among these with approximately 80 died. Symptoms at early stage include common cold、hand-foot-and-mouth disease ( HFMD ) and herpangna. Complication can be fatal as meningitis,myocarditis, poliomyelitis and flaccid paralysis. To advance the anti-enterovirus drug development is, therefore , important to this matter. Various kinds of phycobiliprotein were found being important fluorescent chromatophore in some algae. The objective of the experiment is focused on phycocyanin to study how to inhibit enterovirus 71 activity and its action mechanism. African green monkey kidney cells ( Vero cells ) and Human embryonal rhabdomysarcoma ( RD cells ) were used as materials, and the cytotoxicity test, plaque inhibit assay, neutrolization test, DNA fragmentation assay, cell cycle assay, Annexin-V-Fluos binding assay, and RNA synthsis analysis were used to approach the mechanism. Phycocyanin is found to possess a higher cytotoxicity ( LC50 = 0.129μM ) for Vero cells； while phycoerythrin possesses a medial cytotoxicity ( LC50 = 0.196μM ) and allphycocyanin possesses a lower cytotoxicity ( LC50 = 0.415μM ). By plaque inhibit assay, cells treated with allphycocyanin at the stage of virus adsorption is found to inhibit virus activity better than the ones when allphycocyanin is added after the virus adsorption ( IC50 = 0.056μM ). Phycocyanin and phycoerythrin, on the other hand, do not show significant inhibition of virus activity ( IC50 = 0.062μM and 0.083μM ), either at the stage of virus adsorption or after. By neutrolization test, allphycocyanin can inhibited enterovirus 71 activity while the others phycobiliprotein can not. By DNA fragmentation assay, cell cycle assay, and Annexin-V-Fluos binding assay, it is also found that enterovirus 71 can induce apoptosis in RD cells, while shown no apoptosis in Vero cells. It conjectures that the different tissue cells have the different death by enterovirus 71 infected. By RNA synthsis analysis known, for both the Vero cells and RD cells, allphycocyanin is shown capable of delaying the synthesis of RNA for enterovirus 71. It conjectures that the reason for allphycocyanin inhibited enterovirus 71 infected Vero cells and RD cells is that this.
中文摘要 ---------------------------------------------------iv
英文摘要 ----------------------------------------------------v
誌 謝 ----------------------------------------------------vi
前 言 -----------------------------------------------------1
研究目標 ----------------------------------------------------13
材料與方法----------------------------------------------------14
結 果 ----------------------------------------------------27
討 論 ----------------------------------------------------32
圖 表 ---------------------------------------------------35
參考文獻 ----------------------------------------------------56
附 錄 ---------------------------------------------------70
| Identifer | oai:union.ndltd.org:TW/089CGU00325016 |
| Date | January 2001 |
| Creators | Kun-Nan Tsai, 蔡昆男 |
| Contributors | Err-Cheng Chen, Shin-Rui Shih, 詹爾昌, 施信如 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 94 |
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