研發簡易測量斑馬魚胚胎心律的方法並應用於藥物評估

Development of a simple method to measure cardiac rhythm in zebrafish embryos and its application on drug assessment

沈伯尼(Bonifasius Putera Sampurna)

指導教授 ： 蕭崇德

中原大學/理學院/生物科技研究所/碩士(2019年)

https://doi.org/10.6840/cycu201900095

摘要

在藥物的研發中最重要的就是藥物藥性與毒性的評估，藥物的非專一性的靶向作用可能會引起副作用而致死，其中以心血管毒性是最常發生且受到注意的。藥物誘發的心臟常見毒性有心跳變慢、過快或心房心室脈動不規律。這些化合物在臨床試驗及上市後的潛在毒性可以透過動物實驗來進行活體評估。斑馬魚是一種新興的脊椎動物模式，具產卵數多且胚胎發育快速，在受孕72小時後，心血管就發育完整，可進行早期評估。在發育早期斑馬魚的身體是透明的，因此可從外觀察心血管的發育情形。雖然斑馬魚屬於較原始的脊椎動物，心房室數也不同於人類，但是調節心臟發育與生理的分子作用機制與人類大致相近。斑馬魚的血流速率與心跳率有正相關，當心律改變，血流速率也跟著改變。斑馬魚和人類一樣有閉鎖式的血液循環系統，因此可利用斑馬魚模式來探討人類心血管疾病。之前測量心律及血流速率的方法雖然已被建立，但是其他科學家在進行此實驗時，常遇到的限制是需要使用較為昂貴及複雜的設備，有時還須自己寫程式語言來分析數據。本論文的主要目的是提供一個廉價又簡單的測量心跳及血流速率的設備，可以直接利用ImageJ開源軟體分析未經染色的斑馬魚胚胎，進而獲得心律與血流等珍貴參數。

關鍵字

斑馬魚 ； ImageJ ； 心律 ； 血流

並列摘要

Assessment of the drugs and compounds is the one of the most critical aspect in the drug development. Miss-target property can produce undesired side effects that sometimes lead into lethal consequences. Cardiotoxicity is one of the most common issues. The alteration in the heart beat could be slow down, faster, or irregular rhythm between atrium and ventricle beating. Long QT cases often found in the non-cardiovascular drugs. Animal model could be used to study the detailed process of drug’s side effect, it can be applied in the clinical trial and after marketing phase. Zebrafish is the emerging animal model that has many advantages. Offspring number of zebrafish could reach high amount and the embryonic development process is rapid. After 72hpf heart and blood circulation system already fully developed, observation can be done in early stage. In the initial stage, zebrafish body is transparent making it possible to observe the internal organs including heart from the outside. As a low-level vertebrate, even though the heart chamber number is different from that of human, zebrafish shares the same action potential, molecular, and physiological mechanism of heart like human. Resting heart beat rate of zebrafish is close to human heart beat rate. Blood flow is correlated with heart beating. When the rhythm got altered, the blood flow was also interrupted. Zebrafish has closed blood circulation system same as human, making it possible to use zebrafish as blood model for human disease. Previous methods have been established to calculate the heart rate and blood flow velocity. However, expensive and complicated apparatus is one of the limitation for the other scientists to conduct the experiment. Complicated manual script and programing language is another difficulty. The specific aim of this thesis is to provide low cost and easy setup for heart beat rate and blood flow velocity assessment. By adapting ImageJ as an open source platform, we are able to measure the heart rhythm and blood flow velocity from the wild type zebrafish embryos.

並列關鍵字

Zebrafish ； ImageJ ； heart rate ； blood flow velocity

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