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研究生: 于清華
Yu, Ching-Hua
論文名稱: 以顯微影像分析技術測量奈米銀與奈米銅對斑馬魚心血管功能之影響
Establishing an image-based functional analysis to investigate the effects of silver and copper nanoparticles on cardiovascular system of zebrafish embryos
指導教授: 林豊益
Lin, Li-Yih
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2018
畢業學年度: 107
語文別: 英文
論文頁數: 58
中文關鍵詞: 奈米銀奈米銅斑馬魚心血管功能
英文關鍵詞: AgNP, CuNP, zebrafish, cardiovascular system
DOI URL: http://doi.org/10.6345/THE.NTNU.SLS.001.2019.D01
論文種類: 學術論文
相關次數: 點閱:145下載:0
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  • 奈米材料由於其特性有別於原本的大分子型態,自二十世紀發現後至今已被廣泛用於工業上,如作為殺菌介質的奈米銀,以及用於生物、電化學感應器的奈米銅。然而,對於這些奈米材料可能帶來的環境汙染與對生物體的危害,仍缺乏完整的認識。本研究以斑馬魚的胚胎作為動物模式觀察奈米銀、奈米銅粒子的毒性。在浸泡處理四天後,觀察胚胎的死亡率、孵育率、體長的影響,並透過我們發展的高格數與高畫質顯微攝影技術,測量胚胎心血管功能的受損情形。實驗結果發現,奈米銀與奈米銅在3 ppm就會對上述生理指標有顯著的危害。在心血管功能測量上,發現奈米銀在1 ppm就會導致心輸出量的下降,而3 ppm處理會更進一步地影響心室的收縮力與心率;奈米銅則在0.1 ppm的濃度下就會引發心室體積的減少,在3 ppm處理下還會造成收縮力的下降,而這些奈米金屬引發的心室功能的下降導致了動脈血流速度減緩。綜合以上結果,我們證實了奈米銀、奈米銅對生物體不容忽視的毒性威脅;同時,也驗證了分析心血管功能的顯微攝影技術,在毒理實驗中提供了更敏感的生理指標。

    Found in the 20th century, nanoparticles have been widely used in modern industry due to its exactly different properties compared with the larger particle form of the same composition. However, there is still a lack of thorough understanding of the potential risk that these nanoparticles would bring about. In this study, we assessed the toxicity of AgNP and CuNP by immersing zebrafish embryos in AgNP- or CuNP-containing solution from 4 hr post fertilization (hpf) to 96 hpf. A significant decrease in survival rate, hatching, and body length shows the severe toxicity of AgNP and CuNP. With the application of the videomicroscopic method, which equipped microscopy with a high-frame-rate and high-resolution camera, we found that AgNP cause cardiac output decreased in 1 ppm exposure, and further induced bradycardia and decline in contractility in 3 ppm treated group; on the other hand, a significant shrinkage in ventricular volume was found in 0.1 ppm CuNP treated embryos, and contractility decreased as well in 3ppm exposure. The defects in ventricular contractility caused by two nanoparticles led to a serious decline in the flow velocity of dorsal aorta. To sum up, we suggest that AgNP and CuNP pose a serious threat to the aquatic organism. We also confirmed that this videomicroscopy approach provides a sensitive assessment of the cardiovascular function as a physiological indicator.

    致謝 3 摘要 4 Abstract 5 Introduction 6 Nanoparticles 6 Zebrafish embryos as a model of cardiovascular function assay 7 Purpose 10 Experimental Design 11 Material and Method 12 Experimental animal culture 12 Drug administration 12 Image acquisition 13 Cardiovascular function assessment 13 Analysis of vascular parameters 14 Data analysis 15 Result 16 Inhibitory effect of verapamil on zebrafish embryos’ CV function 16 The survival, hatching and body length of zebrafish embryos after AgNP or CuNP exposure 16 Effects of AgNP on CV function of the zebrafish embryos 18 Effects of CuNP on CV function of the zebrafish embryos 19 Discussion 21 Using the videomicroscopic technique to conduct a precise investigation in CV function of the teleost 21 Aquatic toxicity of AgNP in the early development of zebrafish larvae 26 Aquatic toxicity of CuNP in the early development of zebrafish larvae 30 Conclusion 32 Figure 33 Fig. 1. Assessment of the cardiac function with high-resolution video recording 33 Fig. 2. Assessment of vascular function with high temporal resolution video recording 34 Fig. 3. Effects of verapamil on the ventricular function of zebrafish embryos 36 Fig .4. Effects of verapamil on the aortic vascular function of zebrafish embryos 37 Fig. 5. Effects of AgNP and CuNP on mortality of zebrafish embryos during 96 h exposure 38 Fig. 6. Effects of AgNP and CuNP on the hatching of embryos at 72 hpf and 96 hpf 39 Fig. 7. Effects of AgNP and CuNP on the body length of zebrafish embryos 40 Fig. 8. Effects of AgNP on the ventricular function of the zebrafish embryos 42 Fig. 9. Effects of AgNP on vascular function of the zebrafish embryos 43 Fig. 10. Effects of CuNP on the ventricular function of the zebrafish embryos 45 Fig. 11. Effects of CuNP on vascular function of the zebrafish embryos 46 Fig. 12. The regression analyses between FVAVE and CO calculated from different formulae 47 Fig. 13. The regression analyses between cardiac and vascular indicators 48 Table 1. Effects of AgNP on the CV function of zebrafish embryos 49 Table 2. Effects of CuNP on the CV function of zebrafish embryos 50 References 51

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