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系統識別號 U0026-3008201621481200
論文名稱(中文) 探討脂質代謝對於白點症病毒複製的重要性
論文名稱(英文) Role of lipid metabolism in white spot syndrome virus replication
校院名稱 成功大學
系所名稱(中) 生物科技研究所
系所名稱(英) Institute of Biotechnology
學年度 104
學期 2
出版年 105
研究生(中文) 鄭丞舜
研究生(英文) Cheng-Shun Cheng
學號 L66034033
學位類別 碩士
語文別 中文
論文頁數 69頁
口試委員 指導教授-王涵青
口試委員-陳逸民
口試委員-陳昌熙
口試委員-彭怡禎
口試委員-張麗冠
中文關鍵字 白點症病毒  瓦氏效應  脂質代謝 
英文關鍵字 Penaeus vannamei  White spot syndrome virus  lipolysis  lipogenesis 
學科別分類
中文摘要 全球蝦類養殖產業近年因白點症病毒 (White spot syndrome virus, WSSV) 造成極為嚴重之產業損失,因此研究此病毒之致病機轉以尋求防疫策略。本實驗室先前發現,在WSSV感染後12小時 (病毒複製時期) ,病毒會誘導宿主產生類似由癌症於脊椎動物中引發之瓦氏效應,並在24小時後 (感染晚期) 開始釋放病毒。而本實驗室之實驗首次發現在白點症病毒複製時期、感染晚期會進行脂質分解以及脂質生合成現象。本實驗以螢光染色方式,觀測WSSV感染過程中所影響的脂質代謝平衡改變,顯示當WSSV感染後會影響到宿主脂滴分布。而於專一性抑制 Etomoxir 以及 C75 分別抑制 β-oxidation 和 Fatty acid synthesis 之活性後,由結果推測WSSV在病毒複製時期,會促使肝胰腺活化脂質分解,脂肪酸釋放至血淋巴中再由主要感染之組織 (血球及胃) 吸收後,進而用於能量產生,當進入WSSV感染之晚期,雖然病毒促使感染組織中脂質分解,然而組織中脂質之總面積上升,以提供病毒合成所需之材料,完成病毒之組裝及釋放。從本研究成果推測,脂質分解與脂質生合成應主要提供晚期病毒顆粒之膜狀結構合成使用,以完成病毒顆粒的組裝 (morphogenesis) 。未來將進一步探討WSSV所需之關鍵脂質,作為新的治療/預防/篩選標的。
英文摘要 Global metabolic changes in WSSV-infected shrimp were recently clarified by us (using proteomics and metabolomics). In infected shrimp, re-routing host metabolism (analogous to the Warburg effect in cancer cells) increased availability of energy and building blocks in host cells at the genome replication stage (12 hours post infection; hpi). Moreover, WSSV switched lipid metabolism of host from lipolysis at 12 hpi to lipogenesis at 24 hpi and used it to complete the viral replication cycle and morphogenesis. At the replication stage (12 hpi), lipolysis induced by WSSV in hepatopancreas released free fatty acids that were rapidly assimilated by WSSV target tissues (e.g. hemocytes and stomach). Lipolysis switched to hemocytes and stomach until lipid in hepatopamcreas was exhausted at a late stage (24 hpi). Furthermore, beta-oxidation may be triggered during WSSV infection in shrimp. We determined that WSSV may trigger lipolysis in various tissues during viral replication, and that released free fatty acids may be absorbed by target tissues. Conversely, WSSV failed to complete its replication cycle after beta-oxidation was inhibited; this phenomenon was also observed following inhibition of fatty acid synthetase (FAS; a key enzyme of lipogenesis), during WSSV infection. Therefore, we inferred that alteration of lipid metabolism might be essential for WSSV virion formation. Our study provided new insights into important changes in host lipid metabolism triggered by an invertebrate virus.
論文目次 中文摘要 I
英文摘要 II
誌謝 VI
目錄 VII
表目錄 X
圖目錄 XI
附圖目錄 XIII
縮寫表 XIV
一、研究背景 1
1-1白點症病毒與養蝦產業之關係 1
1-2白點症病毒之特性與致病機轉 2
1-3瓦氏效應 3
1-4脂質代謝路徑對於癌細胞與病毒之重要性 6
1-5研究目的 7
二、材料與方法 9
2-1實驗病毒、動物來源及感染試驗 9
2-2分析白點症病毒感染後之病毒活性 9
2-3白點症病毒感染後病毒基因體量之分析 14
2-4白點症病毒感染後蝦隻之病毒外套膜與組織脂肪酸之分析 16
2-5利用螢光染劑分析脂質於蝦血球內之分布 19
2-6利用電子顯微鏡分析病毒於感染組織細胞中之分佈 20
2-7利用CPT-1抑制劑Etomoxir及FAS抑制劑C75分析對脂肪酸 之影響 21
2-8統計分析方法 22
三、結果 23
3-1白點症病毒感染後之感染組織分布情況與病毒外套膜脂肪酸之分析 23
3-2感染後蝦隻之脂質代謝之變化 24
3-3利用CPT-1抑制劑抑制β-oxidation分析脂質分解之重要性 26
3-4利用C75抑制劑抑制FAS分析脂質合成之重要性 27
四、討論 30
4-1白點症病毒於感染時期可於感染組織細胞核內累積脂質 30
4-2白點症病毒於感染時期可改變蝦隻體內之脂質代謝 31
4-3白點症病毒驅動之脂質分解現象對於病毒之重要性 33
4-4白點症病毒驅動之脂質合成現象對於病毒之重要性 36
4-5脂質分解與脂質生合成貢獻於病毒顆粒合成之假說 37
參考文獻 39
圖表 48
附錄 65

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