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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 張麗冠(Li-Kwan Chang) | |
| dc.contributor.author | Tsui-Yan Chang | en |
| dc.contributor.author | 張翠豔 | zh_TW |
| dc.date.accessioned | 2021-06-15T04:50:34Z | - |
| dc.date.available | 2011-08-06 | |
| dc.date.copyright | 2010-08-06 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-02 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/45987 | - |
| dc.description.abstract | Epstein-Barr virus (EBV)是人類皰疹病毒，具有潛伏期和溶裂期兩種生活型態。EBV的感染途徑是透過唾液感染上皮細胞和淋巴B細胞，與傳染性單核血球增多症、伯基特淋巴瘤、T細胞淋巴癌、何杰金氏症、胃癌及鼻咽癌等疾病有密切關係。BRLF1和BZLF1是EBV的極早期基因，會轉譯Rta和Zta這兩個轉錄因子進而活化EBV溶裂期的基因表現。目前的抗EBV藥物如acyclovir、ganciclovir及indolocarbazole等主要作用於病毒的DNA聚合酶以抑制病毒的溶裂複製。然而病毒常常會突變而產生抗藥性的突變株，因此需要更新穎的藥物及治療方法。Triterpenoids (三萜類)是由三十個碳元素為骨架之天然化合物，普遍存在於植物或真菌中，具有抗發炎、抗腫瘤、抑制細胞增生及誘導細胞凋亡機制的調節作用。本篇研究的目的是探討triterpenoids的衍生物JYK-MA-37對抑制EBV溶裂期的效果。首先將P3HR1細胞以不同濃度的JYK-MA-37處理，接著並以sodium butyrate (SB)及12-O-tetradecanoylphorbol-13-acetate (TPA)誘導EBV進入溶裂循環，利用西方點墨法與流式細胞分析，結果顯示10 μM的JYK-MA-37有效地抑制EBV的溶裂期蛋白質Rta、Zta及EA-D的表現，其EC50-Rta為11.32 μM，CC50為55 μM。接著，以冷光酵素活性分析測試JYK-MA-37對活化BRLF1與BZLF1啟動子Rp及Zp的影響，結果發現EBV溶裂期極早期基因BRLF1及BZLF1的轉錄活性會被抑制。利用即時聚合酶連鎖分析JYK-MA-37對EBV顆粒產生的影響，結果發現JYK-MA-37會抑制EBV顆粒的產生。綜合以上實驗結果，證明JYK-MA-37具有抑制EBV溶裂循環的能力。 | zh_TW |
| dc.description.abstract | Epstein-Barr virus (EBV) is a human herpesvirus, which has two different life cycles: latent and lytic. EBV infects lymphoid and epithelial cells. Infection by this virus causes infectious mononucleosis and is closely related to several malignant diseases, including Burkitt’s lymphoma, T-cell lymphoma, Hodgkin’s disease，gastric cancer and nasopharyngeal carcinoma. At the onset of the EBV lytic cycle, the virus expresses two immediate-early genes, BRLF1 and BZLF1, which encode transcription factors, Rta and Zta, respectively. These two transcription factors are required to activate the EBV early genes and lytic cascade. Clinically effective anti-EBV drugs that target EBV-encoded DNA polymerase, including acyclovir, ganciclovir and indolocarbazole, are commonly used for inhibiting the lytic cycle of EBV. However, viruses are potentially mutagenic for resistance to drugs. In light of this, it is necessary to identify new targets for antivirus chemotherapy and to develop new treatment strategies. Triterpenoids are formed from six isoprene units with 30 carbons. As it is generally known, triterpenoids are major constituents in several herbal remedies or fungi that are reported to have the capacity of anti-inflammatory, anticancer, repressing cell proliferation, inducing cell apotosis program, etc. The purpose of this study is to demonstrate the effect of JYKMA-37, one of the derivatives of triterpenoids, on repressing the EBV lytic cycle. First of all, P3HR1 cells were treated with different concentrations of JYK-MA-37, and then induced the EBV lytic cycle by sodium butyrate (SB) and 12-O-tetradecanoylphorbol-13-acetate (TPA). Immunoblot analysis and flow cytometry analysis were performed, the results showed that JYK-MA-37 at 10 μM effectively inhibits the expression of EBV lytic protein, including Rta、Zta and EA-D, after lytic induction in P3HR1 cells (EC50 = 22.083 μM，CC50 = 11.32 μM). Moreover, transient transfection analysis revealed that transactivation of BRLF1 promoter (Rp) and BZLF1 promoter (Zp) were inhibited by JYK-MA-37 in a dose-dependent manner. Finally, real-time PCR showed that JYK-MA-37 substantially reduces the numbers of EBV particles produced by the cells after lytic induction. Taken together, this study demonstrated that JYK-MA-37 poccess the capacity of inhibiting the progression of EBV lytic cycle. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T04:50:34Z (GMT). No. of bitstreams: 1 ntu-99-R97b47413-1.pdf: 2633448 bytes, checksum: c4124b170cd4c3f37c2b61b597f4503f (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 目錄
口試委員會審定書致謝感言 i 致謝感言 ii 中文摘要 iii 英文摘要 iv 表目錄 ix 圖目錄 x 前言 1 EBV的發現 1 EBV的構造與基因體 1 EBV感染人類的途徑 1 與EBV相關的疾病 2 EBV的生活史 2 極早期基因BRLF1及其Rta蛋白質的表現 4 極早期基因BZLF1及其Zta蛋白質的表現 5 抗皰疹病毒藥物 6 抗EBV活化疾病的治療 8 天然藥物 9 三萜類 (Triterpenoids) 9 Moronic acid (MA)及MA衍生物 10 JYK-MA-37 11 研究目的 12 材料與方法 13 細胞株及培養 13 細胞毒性分析 13 病毒溶裂循環的誘導 13 西方點墨法 13 50% cytotoxic concentration 14 50% effective concentration 14 Therapeutic index 15 流式細胞儀分析 15 質體DNA的萃取 15 RNA的萃取 16 反轉錄聚合酶連鎖反應 16 細胞的轉染 17 冷光酵素活性檢測 17 分離EBV的DNA 17 即時聚合酶連鎖分析 18 結果 19 抗EBV活性藥物的篩選 19 JYK-MA- 37對P3HR1細胞的毒性分析 19 JYK-MA-37對EBV溶裂期蛋白質的影響 20 利用流式細胞儀分析JYK-MA-37對Rta，Zta，EA-D及EBNA1表現的影響 20 JYK-MA-37對BRLF1-BZLF1雙基因轉錄的影響 21 分析JYK-MA-37對EBV極早期基因啟動子Rp與Zp的影響 21 JYK-MA-37對EBV病毒顆粒產生的影響 22 討論 24 參考文獻 46 附件 57 附件一：三萜類的分類及結構 57 附件二：moronic acid的化學結構修改對照表 69 附件三：報導質體 73 | |
| dc.language.iso | zh-TW | |
| dc.title | Triterpenoids抑制Epstein-Barr virus進入溶裂循環 | zh_TW |
| dc.title | Inhibition of Epstein-Barr virus lytic cycle by triterpenoids | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 張芳榮(Fang-Rong Chang),張世宗(Shih-Chung Chang),劉世東(Shi-Tung Liu) | |
| dc.subject.keyword | Epstein-Barr virus (EBV),三萜,類,溶裂循環, | zh_TW |
| dc.subject.keyword | Epstein-Barr virus (EBV),triterpenoids,lytic cycle, | en |
| dc.relation.page | 73 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-08-02 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| Appears in Collections: | 微生物學科所 | |
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| ntu-99-1.pdf Restricted Access | 2.57 MB | Adobe PDF |
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