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系統識別號 U0026-1808202020142000
論文名稱(中文) 動物實驗、替代測試和預測毒理學的比較,以斑馬魚及其胚胎急性毒性試驗與QSAR Toolbox為例
論文名稱(英文) The comparison of animal test, alternative test and predictive toxicology, taking zebrafish (Danio rerio) and its embryo acute toxicity test and QSAR Toolbox as an example
校院名稱 成功大學
系所名稱(中) 環境醫學研究所
系所名稱(英) Institute of Environmental and Occupational Health
學年度 108
學期 2
出版年 109
研究生(中文) 薛建豐
研究生(英文) Jian-Feng Hsueh
學號 S76071067
學位類別 碩士
語文別 中文
論文頁數 116頁
口試委員 指導教授-王應然
口試委員-侯文哲
口試委員-陳容甄
口試委員-童俊維
口試委員-李冠漢
中文關鍵字 魚類急毒性  替代測試  預測毒理學  斑馬魚  定量結構活性關係(QSAR) 
英文關鍵字 Acute fish toxicity test (AFT)  Alternative test  Predictive toxicology  Zebrafish (Danio rerio)  QSAR (Quantitative structure–activity relationship) 
學科別分類
中文摘要 傳統毒理學常常應用動物進行實驗,這樣的研究往往會花費大量金錢及時間,且不符合動物福祉。也因此近年來國際上的趨勢為發展替代測試進而減少金錢及時間,並符合動物福祉,其中一項具有潛力的研究方向為應用定量結構活性關係 (Quantitative structure–activity relationship, QSAR) 模型的預測毒理學。我國近年來對化學物質的管理更加嚴格,廠商需要依照化學物質的安全性以及每年所使用的噸數提供毒理及生態毒理資訊。不過,相較毒理資訊而言,生態毒理資訊的項目卻是相對缺乏的。本研究之目的為驗證及發展能減少使用動物實驗的電腦模式及斑馬魚胚胎替代模式。並且結合電腦模式及斑馬魚胚胎替代模式方法提供魚類急毒性測試終點的整合型測試策略。接著,為了測試整合型測試策略之應用性,以行政院環保署公布的106種優先登錄化學物質中,挑選至少10個物質以QSAR模型來預測毒性,使用電腦軟體為OECD QSAR Toolbox4.4版。然後,操作斑馬魚胚胎急毒性試驗推估成魚急毒性,最後再以成魚急毒性試驗驗證前兩者預測及推估魚類急毒性測試終點之準確性。研究過程中,實驗手法驗證以標準物質3,4-DCA,驗證斑馬魚成魚的魚類急毒性試驗及斑馬魚胚胎急毒性試驗,兩種實驗的分析結果大多都在1個標準差的範圍內,因此本研究通過了兩種實驗的手法驗證。並且完成了十幾種物質的魚類急毒性試驗及斑馬魚胚胎急毒性試驗。在QSAR模型的電腦預測和胚胎的替代測試中,對於成魚急毒性分別有良好的預測及推估能力,而分辨率分別為73.3%和76.6%。最後我們提出了結合兩種替代方法的整合型測試策略,並發現對物質毒性的分辨率提高至93.3%。本研究的結果顯示整合型測試策略能夠減少動物實驗,並以替代測試方法提供生態毒理資訊的魚類急毒性測試終點,也能夠有效率的分類物質的魚類急毒性。
英文摘要 We often utilize animal tests in traditional toxicology, such researches cost lots of money and time. In addition, they are not aligned with animal welfare. The international trend is to develop alternative tests to reduce money and time, which can reduce utilizing animals and close to animal welfare. For our concern of ecotoxicology endpoint, acute fish toxicity test (AFT), is a traditional animal test. There are two alternative tests trying to reduce AFT test, in vitro and in silico, respectively. For in vitro, fish embryo toxicity test is announced as a standard protocol, OECD TG236. For in silico, one of the predictive toxicology methods is to utilize quantitative structure-activity relationship (QSAR) model. In recent years, our country has stricter management of chemical substances and must provide information on the (eco)toxicity of substances by the tonnage of utilization. However, we found that it lacks of ecotoxicology than toxicology information. The objective is to validate and develop methods that can reduce the use of animal tests with zebrafish embryo alternative tests and by computer models. Then, the zebrafish embryo alternative tests and computer model methods are combined to provide an integrated testing strategy (ITS) of AFT test endpoint. The ITS is combined by OECD QSAR Toolbox version 4.4 and OECD TG236. To develop the ITS, the study considered proper experiment without any animal tests. Moreover, we would choose some substances operate the AFT test to validate the ITS results.
論文目次 第一章、序論.........1
第二章、文獻回顧.........2
第一節、傳統毒理學與替代測試.........2
第二節、化學物質的生態毒理資訊.........5
第三節、生態毒理的魚類急毒性及其替代方法.........9
第四節、電腦預測毒理學及定量結構活性關係.........11
第五節、整合型測試策略 (Integrated Testing Strategies, ITS).........18
第三章、研究目的.........23
第四章、研究架構.........24
第五章、研究材料與方法.........25
第一節、從資料庫收集驗證ITS的化學物質.........25
第二節、化學物質的篩選方法.........27
第三節、斑馬魚的飼養方法.........29
第四節、斑馬魚的繁殖方法.........30
第五節、斑馬魚成魚急毒性試驗 (Acute Fish Toxicity Test, AFT Test).........32
第六節、斑馬魚胚胎急毒性試驗 (Zebrafish Embryo Toxicity Test, FET Test).........33
第七節、使用QSAR Toolbox建立QSAR模型.........34
第八節、ITS分類物質的方法.........38
第九節、統計方法.........39
第六章、研究結果.........40
第一節、整合型測試策略之圖示.........40
第二節、由先前資料庫選出的30個化學物質.........40
第三節、斑馬魚成魚急毒性和預測毒理學結果之相關性.........41
第四節、斑馬魚成魚和斑馬魚胚胎急毒性試驗之相關性.........41
第五節、整合性測試策略在已知魚類急毒性化學物質之表現.........42
第六節、使用QSAR模型預測斑馬魚成魚急毒性.........42
第七節、驗證斑馬魚成魚及胚胎急毒性試驗.........43
第八節、動物實驗、替代測試和預測毒理學之結果比較.........43
第九節、整合性測試策略在未知魚類急毒性化學物質之表現.........45
第十節、以生態毒理學水生毒性範圍分類物質毒性.........45
第七章、討論.........46
第一節、動物實驗、替代測試和預測毒理學的比較.........46
第二節、QSAR具有良好的物質分辨能力.........47
第三節、為什麼本研究的ITS可以有良好的預測結果.........47
第四節、ITS在「對魚類有毒性」的物質較為準確.........48
第五節、ITS的限制性.........48
第六節、ITS是否符合WOE,在法規毒理學上的應用.........49
第七節、106種應優先登錄物質的重要性.........50
第八節、化學物質的結構與潛在的毒性.........51
第八章、結論.........52
第九章、參考文獻.........53
圖表.........58
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