跳到主要內容

臺灣博碩士論文加值系統

(44.200.169.3) 您好!臺灣時間:2022/12/05 19:48
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

: 
twitterline
研究生:林珥卿
研究生(外文):Lin Ear-Ching
論文名稱:蘋果汁抑制腸病毒71型複製
論文名稱(外文):Inhibition of enterovirus 71 replication by apple juice
指導教授:吳正男吳正男引用關係林一郎林一郎引用關係
指導教授(外文):Cheng Nan WuElong Lin
學位類別:碩士
校院名稱:中台醫護技術學院
系所名稱:醫學生物科技研究所
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:67
中文關鍵詞:腸病毒71型蘋果汁
外文關鍵詞:enterovirus 71apple juice
相關次數:
  • 被引用被引用:0
  • 點閱點閱:279
  • 評分評分:
  • 下載下載:44
  • 收藏至我的研究室書目清單書目收藏:2
腸病毒71型(Enterovirus 71 , EV71)為一重要的人類致病性病毒,在病毒學上的分類是屬於微小病毒科(Picornaviridae)中之腸病毒屬(enterovirus),為正向單股RNA病毒(positive single strand RNA virus)。腸病毒71型最早於1969年美國加州分離出來,此後在世界各地皆有流行,腸病毒71型在臨床上除手口足病(hand-foot-mouth disease)外,伴隨著中樞神經系統(central nervous system , CNS)疾病,例如心肌炎和腦炎,台灣地區曾在1998年爆發腸病毒大流行造成405名孩童感染重症、78名孩童死亡,到目前為止台灣地區每年都有腸病毒流行,最主要的流行病毒株為腸病毒71型,目前針對腸病毒71型尚無有效的藥物或疫苗可以預防或治療,所以本研究嘗試由天然食材中著手搜尋能有效抑制腸病毒之食材,經多種樣品的測試後發現蘋果汁具有抑制病毒的能力,因此探討其抑制機制為何。由腸病毒71型感染細胞前或感染細胞後分別加入或不加入100倍稀釋的蘋果汁對病毒生長曲線影響之初步實驗中,可見到病毒數量受到稀釋蘋果汁抑制而減少10-100倍的變化,確定蘋果汁具有抑制腸病毒71型複製的能力,同時也在腸病毒71型感染後的細胞存活度、病毒顆粒的釋放、病毒RNA及蛋白質表現量和都可得到類似的結果。由此可知,蘋果汁中確實具有抑制腸病毒71型的功效;且由病毒吸附細胞實驗得知,蘋果汁抑制腸病毒71型的能力可能來自於蘋果汁干擾病毒侵入細胞的早期階段。 同時也初步證實蘋果汁可廣泛抑制多種腸病毒,包括克沙奇B3、B4與B5型。
Enterovirus 71 (EV71) is an important human pathogenic virus. Enterovirus 71 is a positive single strain RNA virus is a member of the Enterovirus genus of the Picornaviridae family. Two EV71 outbreaks resulting in rapid clinical deterioration and death among young children; one outbreak occurred in Bulgaria during May–September 1975 and another in Malaysia during April–June 1997. EV71 infection can cause hand-foot-mouth disease (HFMD) is following the central nervous system (CNS), the disease, such as the myocarditis and encephalitis. In 1998 there was a large HFMD outbreak be heavy to cause 405 child infect with serious disease, 78 child death. EV71 is the most pathogenic virus in acute enterovirus complication in Taiwan till now. Effective medicine or vaccine that can be prevented or treated for EV71 is not yet available at present. This research tries to be set about searching natural food can inhibits EV71 effectively. After test of many kinds of samples, the apple juice it has been shown to have anti-viral activity. The apple juice inhibits EV71 virus replication and cytopathic effect in cell culture. Confirm it has ability to inhibit EV71 virus replication from the cell survival, viral RNA expression, viral protein expression, and viral particle release experiments. Therefore, the apple juice really has ability to blocked cytopathic effect and inhibits the EV71 viral replication. Further, we found the apple juice effectively blocked EV71 viral attachment to the cell from the experiment of attachment assay. Verify tentatively, the apples juice can also inhibit extensively enterovirus, including Coxsackievirus B3, Coxsackievirus B4 and Coxsackievirus B5.
摘要 I
目錄 2
第一章 前言 5
第二章 文獻回顧 3
第一節 腸病毒之特性分析 4
一、 病毒分類與流行史 4
二、 病毒顆粒與基因體組成 5
三、 病毒生活史 5
第二節 蘋果之特性分析 6
一、 蘋果之簡介 6
二、 蘋果之生理功能 7
第三節 研究目標 8
第一節 實驗材料 10
一、 病毒及細胞株 10
第二節 實驗方法 13
一、 細胞培養 13
二、 病毒增生 13
三、 蘋果汁製備 13
四、 蘋果汁抑制腸病毒試驗 14
五、 病毒生長曲線分析及定量 14
六、 以LDH assay測量細胞存活率 15
七、 胞內病毒RNA萃取與半定量 16
八、 胞內病毒蛋白質萃取及定量 18
九、免疫螢光染色 20
十、病毒吸附試驗 20
十一、病毒斑免疫分析 21
第四章 實驗結果 23
第一節 蘋果汁抑制腸病毒71型感染 24
第二節 蘋果汁減緩細胞被腸病毒71型感染所造成的死亡 24
第三節 蘋果汁影響細胞內腸病毒71型 RNA的表現 25
第四節 蘋果汁影響細胞內腸病毒71型蛋白質量的變化 26
第五節 蘋果汁對腸病毒71型吸附細胞的影響 27
第六節 蘋果汁廣泛抑制腸病毒流行株感染 28
第七節 蘋果汁廣泛減緩腸病毒感染細胞所造成的死亡 29
第五章 討論與結論 30
討論 31
結論 36
第六章 參考文獻 37
圖表 42
表一、蘋果營養成分表 43
圖二、蘋果汁抑制以高劑量腸病毒71型感染後的複製 45
圖三、 蘋果汁添加時間點對抑制腸病毒71型感染效果的比較 46
圖四、蘋果汁對腸病毒71型感染細胞後,細胞存活率的影響 47
圖五、蘋果汁干擾細胞內腸病毒71型 RNA的表現 48
圖六、以西方墨點法分析蘋果汁影響腸病毒71型感染細胞後的蛋白質變化量 49
圖七、蘋果汁降低細胞內腸病毒71型病毒的產生 50
圖八、蘋果汁添加時間點對細胞內腸病毒71型病毒產生的影響 51
圖九、蘋果汁影響腸病毒71型吸附到細胞表面 52
圖十、蘋果汁抑制克沙奇B3感染後複製 53
圖十一、蘋果汁抑制克沙奇B4感染後複製 54
圖十二、蘋果汁抑制克沙奇B5感染後複製 55
圖十三、蘋果汁對廣泛的台灣流行腸病毒株感染細胞後,細胞存活率的影響 56
附錄 57
AbuBakar, S., H. Y. Chee, et al. (1999). "Identification of enterovirus 71 isolates from an outbreak of hand, foot and mouth disease (HFMD) with fatal cases of encephalomyelitis in Malaysia." Virus Res 61(1): 1-9.
Akazome, Y. (2004). "Characteristics and physiological functions of polyphenols from apples." Biofactors 22(1-4): 311-4.
Aprikian, O., V. Duclos, et al. (2003). "Apple pectin and a polyphenol-rich apple concentrate are more effective together than separately on cecal fermentations and plasma lipids in rats." J Nutr 133(6): 1860-5.
Buenz, E. J. and C. L. Howe (2006). "Picornaviruses and cell death." Trends Microbiol 14(1): 28-36.
Chumakov, M., M. Voroshilova, et al. (1979). "Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria." Arch Virol 60(3-4): 329-40.
Egger, D. and K. Bienz (2005). "Intracellular location and translocation of silent and active poliovirus replication complexes." J Gen Virol 86(Pt 3): 707-18.
Gacche, R. N., G. B. Zore, et al. (2003). "Kinetics of inhibition of polyphenol oxidase mediated browning in apple juice by beta-cyclodextrin and L-ascorbate-2-triphosphate." J Enzyme Inhib Med Chem 18(1): 1-5.
Gauillard, F., F. Richard-Forget, et al. (1993). "New spectrophotometric assay for polyphenol oxidase activity." Anal Biochem 215(1): 59-65.
Gosse, F., S. Guyot, et al. (2005). "Chemopreventive properties of apple procyanidins on human colon cancer-derived metastatic SW620 cells and in a rat model of colon carcinogenesis." Carcinogenesis 26(7): 1291-5.
Graziani, G., G. D'Argenio, et al. (2005). "Apple polyphenol extracts prevent damage to human gastric epithelial cells in vitro and to rat gastric mucosa in vivo." Gut 54(2): 193-200.
Guyot, S., N. Marnet, et al. (2003). "Variability of the polyphenolic composition of cider apple (Malus domestica) fruits and juices." J Agric Food Chem 51(21): 6240-7.
Hagiwara, A., I. Tagaya, et al. (1978). "Epidemic of hand, foot and mouth disease associated with enterovirus 71 infection." Intervirology 9(1): 60-3.
Hamauzu, Y., H. Yasui, et al. (2005). "Phenolic profile, antioxidant property, and anti-influenza viral activity of Chinese quince (Pseudocydonia sinensis Schneid.), quince (Cydonia oblonga Mill.), and apple (Malus domestica Mill.) fruits." J Agric Food Chem 53(4): 928-34.
Ho, M. (2000). "Enterovirus 71: the virus, its infections and outbreaks." J Microbiol Immunol Infect 33(4): 205-16.
Hsiung, G. D. and J. R. Wang (2000). "Enterovirus infections with special reference to enterovirus 71." J Microbiol Immunol Infect 33(1): 1-8.
Kahle, K., M. Kraus, et al. (2005). "Polyphenol profiles of apple juices." Mol Nutr Food Res 49(8): 797-806.
Konowalchuk, J. and J. I. Speirs (1978). "Antiviral effect of apple beverages." Appl Environ Microbiol 36(6): 798-801.
Konowalchuk, J. and J. I. Speirs (1978). "Antiviral effect of commercial juices and beverages." Appl Environ Microbiol 35(6): 1219-20.
Kubo, J., J. R. Lee, et al. (1999). "Anti-Helicobacter pylori agents from the cashew apple." J Agric Food Chem 47(2): 533-7.
Li, X. L., J. A. Blackford, et al. (1998). "RNase L mediates the antiviral effect of interferon through a selective reduction in viral RNA during encephalomyocarditis virus infection." J Virol 72(4): 2752-9.
Lin, K. H., K. P. Hwang, et al. (2006). "Evolution of EV71 genogroup in Taiwan from 1998 to 2005: an emerging of subgenogroup C4 of EV71." J Med Virol 78(2): 254-62.
Lin, T. Y., S. J. Twu, et al. (2003). "Enterovirus 71 outbreaks, Taiwan: occurrence and recognition." Emerg Infect Dis 9(3): 291-3.
Liu, C. C., H. W. Tseng, et al. (2000). "An outbreak of enterovirus 71 infection in Taiwan, 1998: epidemiologic and clinical manifestations." J Clin Virol 17(1): 23-30.
Muir, P. and A. M. van Loon (1997). "Enterovirus infections of the central nervous system." Intervirology 40(2-3): 153-66.
Pelletier, I., G. Duncan, et al. (1998). "Molecular mechanisms of poliovirus persistence: key role of capsid determinants during the establishment phase." Cell Mol Life Sci 54(12): 1385-402.
Pietiainen, V. M., V. Marjomaki, et al. (2005). "Viral entry, lipid rafts and caveosomes." Ann Med 37(6): 394-403.
Shekhar, K., M. S. Lye, et al. (2005). "Deaths in children during an outbreak of hand, foot and mouth disease in Peninsular Malaysia--clinical and pathological characteristics." Med J Malaysia 60(3): 297-304.
Shimizu, H., A. Utama, et al. (1999). "Enterovirus 71 from fatal and nonfatal cases of hand, foot and mouth disease epidemics in Malaysia, Japan and Taiwan in 1997-1998." Jpn J Infect Dis 52(1): 12-5.
van der Sluis, A. A., M. Dekker, et al. (1997). "Flavonoids as bioactive components in apple products." Cancer Lett 114(1-2): 107-8.
Wang, S. M., C. C. Liu, et al. (1999). "Clinical spectrum of enterovirus 71 infection in children in southern Taiwan, with an emphasis on neurological complications." Clin Infect Dis 29(1): 184-90.
Wolfe, K., X. Wu, et al. (2003). "Antioxidant activity of apple peels." J Agric Food Chem 51(3): 609-14.
Yan, J. J., J. R. Wang, et al. (2000). "An outbreak of enterovirus 71 infection in Taiwan 1998: a comprehensive pathological, virological, and molecular study on a case of fulminant encephalitis." J Clin Virol 17(1): 13-22.
梁佑慎,柯立祥(2002),市售新鮮蔬果抗氧化力之研究,屏東科技大學
熱帶農業暨國際合作研究所。
李雅志,許明仁(2000),臭氧及溶液處理對輕度加工蘋果楔形切塊品質改善之研究 ,國立臺灣大學園藝學研究所。
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top