臺灣博碩士論文加值系統

萊克多巴胺 (Ractopamine, RAC) 是一種 β-腎上腺素受體激動劑，用於促進牲畜的瘦肉和提高食物轉化效率。這種化合物被認為會導致家畜（如豬）的行為和生理發生改變。很少有研究探討 RAC 在水生動物中的潛在非目標效應。在本研究中，我們通過評估運動活力、耗氧量和心血管功能等多個參數，探索斑馬魚急性暴露RAC後的生理反應。斑馬魚幼魚在 0.1、1、2、4 或 8 ppm 的RAC 暴露 24 小時，並監測和量化相對應的心血管、呼吸和運動活力。此外，我們還對 RAC 與 10 種斑馬魚內源性 β-腎上腺素受體進行了分子對接，以闡明 RAC 的作用機制。結果表明，RAC 可以顯著提高運動活力、心臟性能、耗氧量和血流量，但不會影響斑馬魚胚胎的心律規律。基於結構的分子對接，RAC 對所有 10 種內源性 β-腎上腺素受體亞型（從 adra1aa 到 adra2db）都表現出相似的結合親和力，與人類相同。這一結果表明 RAC 可作為高效、廣譜的 β-腎上腺素受體激動劑，促進斑馬魚的運動活力、心臟功能和耗氧量。為了驗證我們的結果，我們使用β-受體阻滯劑(Propranolol, PROP)與 RAC 共同培養。 PROP 暴露可以減少斑馬魚幼魚的運動過度活躍、高耗氧量和高心率。基於計算機結構的分子模擬和結合親和力測試表明，PROP 的結合親和力總體上低於 RAC。總之，我們的研究提供了可靠的活體證據，支持 RAC可以調節斑馬魚的心血管、呼吸和運動生理。

Ractopamine (RAC) is a beta-adrenoceptor agonist that is used to promote lean and increased food conversion efficiency in livestock. This compound has been considered to be causing behavioral and physiological alterations in livestock like pig. Few studies have addressed the potential non-target effect of RAC in aquatic animals. In this study, we aimed to explore the potential physiological response after acute RAC exposure in zebrafish by evaluating multiple end-points like locomotor activity, oxygen consumption, and cardiovascular performance. Zebrafish larvae were subjected to waterborne RAC exposure at 0.1, 1, 2, 4, or 8 ppm for 24 h, and the corresponding cardiovascular, respiratory, and locomotion activities were monitored and quantified. In addition, we also performed in silico molecular docking for RAC with 10 zebrafish endogenous β-adrenergic receptors to elucidate the potential acting mechanism of RAC. Results show RAC administration can significantly boost locomotor activity, cardiac performance, oxy-gen consumption, and blood flow rate, but without affecting the cardiac rhythm regularity in zebrafish embryos. Based on structure-based flexible molecular docking, RAC display similar binding affinity to all ten subtypes of endogenous β-adrenergic receptors, from adra1aa to adra2db, which are equivalent to the human one. This result suggests RAC might act as high potency and broad spectrum β-adrenergic receptors agonist on boosting the locomotor activity, cardiac performance, and oxygen consumption in zebrafish. To validate our results, we co-incubated a well-known β-blocker of propranolol (PROP) with RAC. PROP exposure tends to minimize the locomotor hyperactivity, high oxygen consumption, and cardiac rate in zebrafish larvae. In silico structure-based molecular simulation and binding affinity tests show PROP has an overall lower binding affinity than RAC. Taken together, our studies provide solid in vivo evidence to support that RAC plays crucial roles on modulating cardiovascular, respiratory, and locomotory physiology in zebrafish for the first time. In addition, the versatile functions of RAC as β-agonist possibly mediated via receptor competition with PROP as β-antagonist.

CONTENTS

中文摘要 ……………………………………………………………………………………..I
ABSTRACT II
ACKNOWLEDGEMENTS III
CONTENTS IV
LIST OF FIGURES V
LIST OF TABLES V
Evaluation of Ractopamine on Cardiovascular, Respiratory and Locomotory Physiology in Zebrafish Larvae………………………………………………………..1-24

INTRODUCTION 1
MATERIALS AND METHODS 3
RESULTS 7
DISCUSSION 14
REFERENCES 32


LIST OF FIGURES
Evaluation of Ractopamine on Cardiovascular, Respiratory and Locomotory Physiology in Zebrafish Larvae…………………………………………………………18-27

Figure 1 18
Figure 2 19
Figure 3. 20
Figure 4 21
Figure 5 22
Figure 6 23
Figure 7.. 24
Figure 8 25
Figure 9. 26
Figure 10.. 27


LIST OF TABLES

Evaluation of Ractopamine on Cardiovascular, Respiratory and Locomotory Physiology in Zebrafish Larvae………………......……………………………….28-30
Table 1. 28
Table 2 29
Table 3 30

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