Laboratory of Analytical Chemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece.
Department of Biological Applications and Technologies, University of Ioannina, Ioannina, Greece.
Methods Mol Biol. 2024;2753:495-502. doi: 10.1007/978-1-0716-3625-1_31.
In order for new drugs to enter the market, extensive studies are needed to examine toxic effects. Among others, teratogenicity studies are of paramount importance. Of even higher importance is to gain knowledge on the biological responses that take place upon drug exposure, so as to have a better understanding of the molecular mechanisms that govern developmental changes. Metabolomics is the research field that studies the changes in the chemical composition of metabolites contained within cells. Conducting metabolomics studies results in valuable information. Zebrafish is a vertebrate model organism that bridges in vivo assays and in vivo studies. In this chapter, we propose a metabolomic fingerprint assay for the study of metabolic changes in zebrafish embryos upon exposure to various drugs. The metabolome of zebrafish is extracted, and the H-NMR spectrum is recorded. Using open-access metabolomic databases, a list of tentative metabolites is retrieved. The presence of the tentative metabolites is further confirmed by UHPLC-HRMS. Ultimately, after a metabolic pathway analysis, the metabolic network is revealed and useful conclusions can be drawn.
为了让新药进入市场，需要进行广泛的研究来检查其毒性作用。其中，致畸性研究至关重要。更重要的是要了解药物暴露后发生的生物反应，以便更好地理解控制发育变化的分子机制。代谢组学是研究细胞内代谢物化学成分变化的研究领域。进行代谢组学研究可以获得有价值的信息。斑马鱼是一种脊椎动物模式生物，可将体内测定与体内研究联系起来。在本章中，我们提出了一种代谢组指纹分析方法，用于研究暴露于各种药物后斑马鱼胚胎的代谢变化。提取斑马鱼的代谢组，记录 H-NMR 图谱。使用开放访问的代谢组学数据库，检索到暂定代谢物的列表。通过 UHPLC-HRMS 进一步确认暂定代谢物的存在。最终，在进行代谢途径分析后，揭示代谢网络，并得出有用的结论。
