Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 08826, Republic of Korea.
School of Applied Biosciences, Kyungpook National University, Daegu, 41566, Republic of Korea.
J Hazard Mater. 2020 May 5;389:122056. doi: 10.1016/j.jhazmat.2020.122056. Epub 2020 Jan 15.
Endosulfan sulfate is a major oxidative metabolite of the chlorinated insecticide endosulfan. In this study, a targeted metabolomics approach was used to investigate the toxic mechanisms of endosulfan sulfate in adult zebrafish using the multiple reaction monitoring mode of a GC-MS/MS. The LC of endosulfan sulfate in adult zebrafish was determined and then zebrafish were exposed to endosulfan sulfate at one-tenth the LC (0.1LC) or the LC for 24 and 48 h. After exposure, the fish were extracted, derivatized and analyzed by GC-MS/MS for 379 metabolites to identify 170 metabolites. Three experimental groups (control, 0.1LC and LC) were clearly separated in PLS-DA score plots. Based on the VIP, ANOVA, and fold change results, 40 metabolites were selected as biomarkers. Metabolic pathways associated with those metabolites were identified using MetaboAnalyst 4.0 as follows: aminoacyl-tRNA biosynthesis, valine/leucine/isoleucine biosynthesis, citrate cycle, glycerolipid metabolism, and arginine/proline metabolism. Gene expression studies confirmed the activation of citrate cycle and glycerolipids metabolism. MDA levels of the exposed group significantly increased in oxidative toxicity assay tests. Such significant perturbations of important metabolites within key biochemical pathways must result in biologically hazardous effects in zebrafish.
硫丹硫酸盐是氯化杀虫剂硫丹的主要氧化代谢物。在这项研究中，使用 GC-MS/MS 的多重反应监测模式，采用靶向代谢组学方法研究了硫丹硫酸盐对成年斑马鱼的毒性机制。测定了成年斑马鱼中硫丹硫酸盐的 LC 值，然后将斑马鱼暴露于 LC 的十分之一（0.1LC）或 LC 下 24 和 48 小时。暴露后，通过 GC-MS/MS 对鱼进行提取、衍生化和分析，以鉴定 379 种代谢物中的 170 种代谢物。PLS-DA 得分图清楚地区分了三个实验组（对照组、0.1LC 和 LC）。根据 VIP、ANOVA 和倍数变化结果，选择了 40 种代谢物作为生物标志物。使用 MetaboAnalyst 4.0 确定与这些代谢物相关的代谢途径如下：氨酰-tRNA 生物合成、缬氨酸/亮氨酸/异亮氨酸生物合成、柠檬酸循环、甘油脂质代谢和精氨酸/脯氨酸代谢。基因表达研究证实了柠檬酸循环和甘油脂质代谢的激活。在氧化毒性测定试验中，暴露组的 MDA 水平显著增加。这些关键生物化学途径中重要代谢物的显著扰动，必然会对斑马鱼产生生物危害性影响。
