Zhu Limei, Gao Na, Wang Ruifang, Zhang Li
Key Laboratory of Tropical Marine Bio-Resources and Ecology, Guangdong Provincial Key Laboratory of Applied Marine Biology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Ecotoxicology. 2018 Apr;27(3):267-277. doi: 10.1007/s10646-017-1892-2. Epub 2018 Jan 10.
Ammonia is both a highly toxic environmental pollutant and the major nitrogenous waste produced by ammoniotelic teleosts. Although the acute toxic effects of ammonia have been widely studied in fish, the biochemical mechanisms of its toxicity have not been understood comprehensively. In this study, we performed comparative proteomic and metabolomic analysis between ammonia-challenged (1.2 and 2.6 mmol L NHCl for 96 h) and control groups of marine medaka (Oryzias melastigma) to identify changes of the metabolite and protein profiles in response to ammonia stress. The metabolic responses included changes of multiple amino acids, carbohydrates (glucose and glycogen), energy metabolism products (ATP and creatinine), and other metabolites (choline and phosphocholine) after ammonia exposure, indicating that ammonia mainly caused disturbance in energy metabolism and amino acids metabolism. The two-dimensional electrophoresis-based proteomic study identified 23 altered proteins, which were involved in nervous system, locomotor system, cytoskeleton assembly, immune stress, oxidative stress, and signal transduction of apoptosis. These results suggested that ammonia not only induced oxidative stress, immune stress, cell injury and apoptosis but also affected the motor ability and central nervous system in marine medaka. It is the first time that metabolomic and proteomic approaches were integrated to elucidate ammonia toxicity in marine fishes. This study is of great value in better understanding the mechanisms of ammonia toxicity in marine fishes and in practical aspects of aquaculture.
氨既是一种剧毒的环境污染物，也是氨排泄型硬骨鱼产生的主要含氮废物。尽管氨对鱼类的急性毒性作用已得到广泛研究，但其毒性的生化机制尚未得到全面了解。在本研究中，我们对氨暴露组（分别用1.2和2.6 mmol/L氯化铵处理96小时）和对照组的海洋青鳉（Oryzias melastigma）进行了比较蛋白质组学和代谢组学分析，以确定氨应激下代谢物和蛋白质谱的变化。代谢反应包括氨暴露后多种氨基酸、碳水化合物（葡萄糖和糖原）、能量代谢产物（三磷酸腺苷和肌酐）以及其他代谢物（胆碱和磷酸胆碱）的变化，表明氨主要导致能量代谢和氨基酸代谢紊乱。基于二维电泳的蛋白质组学研究鉴定出23种变化的蛋白质，它们参与神经系统、运动系统、细胞骨架组装、免疫应激、氧化应激和细胞凋亡信号转导。这些结果表明，氨不仅诱导氧化应激、免疫应激、细胞损伤和凋亡，还影响海洋青鳉的运动能力和中枢神经系统。这是首次将代谢组学和蛋白质组学方法结合起来阐明海洋鱼类的氨毒性。本研究对于更好地理解海洋鱼类氨毒性机制以及水产养殖实际应用具有重要价值。
