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研究生: 馮瑩茜
Phong, Ying-Chiann
論文名稱: N-乙醯半胱氨酸對斑馬魚仔魚側線毛細胞的影響
Effects of N-acetyl cysteine on Lateral Line Hair Cell of Zebrafish Larvae (Danio Rerio)
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: 2019
畢業學年度: 107
語文別: 英文
論文頁數: 46
中文關鍵詞: N-乙醯半胱氨酸抗氧化劑新黴素斑馬魚側線毛細胞耳毒性胚胎毒性
英文關鍵詞: N-acetyl cysteine, antioxidant, neomycin, zebrafish, lateral line, hair cell, ototoxicity, embryotoxicity
DOI URL: http://doi.org/10.6345/THE.NTNU.SLS.005.2019.D01
論文種類: 學術論文
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  • N-乙醯半胱氨酸(N-acetyl cysteine,NAC)是一種作為營養食品的抗氧化劑。在水產養殖上使用N-乙醯半胱氨酸餵食吳郭魚亦可以降低微囊藻毒素和柱孢藻毒素所產生的氧化壓力。此研究的目的是想要探討抗氧化劑NAC的潛在毒性及致死劑量,以便未來在魚類上作為抗氧化劑的應用。此外測試NAC對於新黴素所造成耳毒性傷害是否有保護作用。我們將斑馬魚長時間及短時間曝露於NAC,並觀察NAC對斑馬魚的影響。我們的研究結果顯示NAC會對斑馬魚仔魚造成死亡的現象,並且其藥效與濃度成正相關。另外,NAC對各個發育的指標都會帶來負面的影響,如存活率、孵化率、斑馬魚仔魚的體長及心跳速率。另外,使用FM1-43這活體染劑,可得知NAC在急性或是慢性處理下都會減少斑馬魚仔魚的毛細胞。急性NAC處理後會增加毛細胞的氧化壓力。除此之外,NAC亦會減少斑馬魚仔魚卵黃囊上的離子細胞密度。從以上的結果可得知不管是急性或是慢性的NAC處理都會對斑馬魚造成毒性傷害。

    N-acetyl cysteine (NAC), an antioxidant used as a nutritional supplement, was used to protect tilapia from oxidative stress-induced by Microcystin and Cylindrospermopsin in the aquaculture sector. The aim of the present study was to investigate the potential toxicity and lethal dose of NAC before applying it to be an antioxidant in fish. Furthermore, we test for the protective effect of NAC against neomycin-induced ototoxicity. We investigate the embryonic toxicity of NAC on zebrafish larvae by acute or chronic exposure to NAC. Our results showed that NAC caused a lethal effect on zebrafish embryo in a dose-dependent manner. By the development assessments, survival rate, body length, and heart rate significantly decreased after the NAC treatment. By labeling hair cells with FM1-43, the hair cells number decreased in both acute and chronic NAC treatment groups. Oxidative stress of lateral line hair cell increased after acute NAC treatment. Moreover, NAC reduced the density of ionocyte on zebrafish yolk sac. Our findings demonstrate that NAC may cause toxicity to zebrafish in both acute and chronic treatment.

    Introduction 1 N-acetyl cysteine 1 Ototoxicity 2 Advantages of zebrafish lateral line 3 Zebrafish yolk sac ionocyte 4 Zebrafish model 5 Purpose 6 Experimental design 7 Materials and methods 8 Zebrafish husbandry 8 Drug administration 8 Embryos developmental assessments 9 Staining of hair cells within neuromasts 9 Rhodamine123 staining of ionocyte 9 ROS detection 10 The analysis of ion content in zebrafish larvae 10 Statistical analysis 11 Results 12 N-acetyl cysteine treatment affects zebrafish embryogenesis 12 Chronic treatment of N-acetyl cysteine affects the zebrafish larvae lateral line hair cells number 13 Chronic treatment of N-acetyl cysteine affects zebrafish larvae yolk sac ionocyte density 13 N-acetyl cysteine affects ion contents of zebrafish 14 Acute treatment of N-acetyl cysteine affects zebrafish lateral line hair cell number 14 Acute treatment of N-acetyl cysteine affects zebrafish larvae yolk sac ionocyte density 15 The oxidative stress level of zebrafish lateral line hair cell was increased after acute treatment of N-acetyl cysteine 15 Co-administration of N-acetyl cysteine does not have a protective effect against neomycin-induced hair cell damage 16 Pre-treatment with N-acetyl cysteine does not have a protective effect against neomycin-induced hair cell loss 16 Discussion 18 N-acetyl cysteine induces reductive stress on zebrafish larvae 18 Effect of N-acetyl cysteine on zebrafish larvae ion regulation 19 Effect of N-acetyl cysteine on zebrafish lateral line hair cell 21 Protective effect of NAC against neomycin-induced hair cell damage 22 Conclusion 24 References 25 Figures 31 Figure 1. Effect of N-acetyl cysteine treatment on zebrafish embryogenesis. 31 Figure 2. Effect of N-acetyl cysteine treatment on zebrafish lateral line hair cells. 32 Figure 3. Effect of N-acetyl cysteine treatment on zebrafish yolk sac ionocyte. 33 Figure 4. Effect of N-acetyl cysteine treatment on zebrafish ion content. 34 Figure 5. Effect of N-acetyl cysteine acute treatment on zebrafish lateral line hair cell. 36 Figure 6. Effect of N-acetyl cysteine acute treatment on zebrafish yolk sac ionocyte density. 37 Figure 7. Oxidative stress of N-acetyl cysteine treatment on zebrafish lateral line hair cell. 40 Figure 8. The protective effect of N-acetyl cysteine on neomycin-induced hair cell damage. 41 Figure 9. The protective effect of pre-treatment with N-acetyl cysteine on neomycin-induced hair cell damage. 43

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