Metabolomic Fingerprint of the Model Ciliate, Tetrahymena thermophila Determined by Untargeted Profiling Using Gas Chromatography-Mass Spectrometry
Metabolomic Fingerprint of the Model Ciliate, Tetrahymena thermophila Determined by Untargeted Profiling Using Gas Chromatography-Mass Spectrometry
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DOI：
10.1007/s11802-019-3974-7
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发表时间：
2019-06
影响因子：
1.6
通讯作者：
Jing Xu;Tao Bo;Weibo Song;Wei Wang
中科院分区：
文献类型：
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作者：
Jing Xu;Tao Bo;Weibo Song;Wei Wang
四膜虫是生态毒理学研究的重要模式生物。细胞内代谢的变化是由环境中的外源性化学物质引起的。细胞内代谢物的变化意味着毒性作用，可以通过代谢组学分析进行监测。本研究建立了一套基于GC-MS的四膜虫代谢组学分析方法。不同提取溶剂对嗜热四膜虫代谢组学分析的影响不同。在浸提溶剂配方1（F1）样品中检测到的代谢物峰强度最强且稳定，同时鉴定了61种代谢物。配方1显示出对碳水化合物的优异提取性能。在提取溶剂配方2（F2）的样品中，对66种代谢产物进行了表征，并提取了脂肪酸代谢产物。同时，在用式3（F3）和式4（F4）的提取中分别表征了57和58种代谢物。但代谢产物的峰强度较低，且代谢产物不稳定。这些结果表明，不同的提取溶剂显著影响细胞内代谢物的检测覆盖度和峰强度。在所有实验组中共鉴定出74种代谢产物（19种氨基酸、11种有机酸、2种无机酸、11种脂肪酸、11种碳水化合物、3种糖苷、4种醇、6种胺和7种其他化合物）。其中，氨基酸、甘油、肌醇和不饱和脂肪酸等代谢物有可能成为检测T. thermophila对环境压力。
The ciliateTetrahymenais a valuable model organism in the studies of ecotoxicology. Changes in intracellular metabolism are caused by exogenous chemicals in the environment. Intracellular metabolite changes signify toxic effects and can be monitored by metabolomics analysis. In this work, a protocol for the GC-MS-based metabolomic analysis ofTetrahymenawas established. Different extraction solvents showed divergent effects on the metabolomic analysis ofTetrahymena thermophila. The peak intensity of metabolites detected in the samples of extraction solvent Formula 1 (F1) was the strongest and stable, while 61 metabolites were identified. Formula 1 showed an excellent extraction performance for carbohydrates. In the samples of extraction solvent Formula 2 (F2), 66 metabolites were characterized, and fatty acid metabolites were extracted. Meanwhile, 57 and 58 metabolites were characterized in the extraction with Formula 3 (F3) and Formula 4 (F4), respectively. However, the peak intensity of the metabolites was low, and the metabolites were unstable. These results indicated that different extraction solvents substantially affected the detected coverage and peak intensity of intracellular metabolites. A total of 74 metabolites (19 amino acids, 11 organic acids, 2 inorganic acids, 11 fatty acids, 11 carbohydrates, 3 glycosides, 4 alcohols, 6 amines, and 7 other compounds) were identified in all experimental groups. Among these metabolites, amino acids, glycerol, myoinositol, and unsaturated fatty acids may become potential biomarkers of metabolite set enrichment analysis for detecting the ability ofT. thermophilaagainst environmental stresses.