4-氯-3-甲基苯酚(PCMC)暴露对成年斑马鱼内源性物质的影响

建立了4-氯-3-甲基苯酚（PCMC）的水生生物斑马鱼暴露毒理学模型,研究了斑马鱼PCMC暴露后内源性代谢物的变化.同时,在斑马鱼PCMC半数致死浓度LC₅₀的基础上,以1/10 LC₅₀为暴露浓度进行代谢组学分析.最后,经超高效液相色谱串联三重四级杆飞行时间质谱（UHPLC-Triple-TOF-MS）检测,结合主成分分析（PCA）和正交偏最小二乘法判别分析（OPLS-DA）进行代谢谱分析.结果发现,空白对照组和暴露组在代谢水平上有明显差异.进一步分析后筛选出9种上调物质和11种下调物质作为潜在生物标志物,主要包括氨基酸、嘌呤、糖类和脂质.根据差异代谢物生理功能分析发现,PCMC主要通过影响嘌呤代谢、甘油磷脂代谢、淀粉和蔗糖代谢、组氨酸代谢、丙酮酸代谢及苯丙氨酸、酪氨酸和色氨酸的生物合成,对斑马鱼的心血管系统、神经系统、蛋白代谢和能量代谢等方面产生毒性效应.本研究建立的代谢组学方法能全面评价氯酚类消毒剂对水生生物的毒性效应,为进一步阐明氯酚类消毒剂的毒性机制提供了理论基础.

The effect of 4-chloro-3-methylphenol(PCMC) on endogenous substances in adult zebrafish(Danio rerio)

In this paper, 4-chloro-3-methylphenol (PCMC) was applied for zebrafish to study the changes in endogenous metabolites. According to the lethal concentration 50% (LC₅₀) of zebrafish PCMC exposure, the concentration of 10% LC₅₀ was performed for metabolomics analysis. Ultra-high performance liquid chromatography coupled with triple time of flight mass spectrometer (UHPLC-Triple-TOF-MS) was used to analyze the effects of PCMC on endogenous substances in zebrafish. The principal components analysis (PCA) and supervised orthogonal partial lease squares discriminant method (OPLS-DA) were used to analyze the metabolic profile. The results showed that metabolic differences were significant in PCMC exposure compared with the blank control group. 9 up-regulated substances and 11 down-regulated substances were selected as potential biomarkers which included amino acids, purines, sugars and lipid. Based on the physiological function of different metabolites, the toxin effect of PCMC on cardiovascular, nervous, protein metabolism and energy metabolism were involved in purine metabolism, glycerophospholipid metabolism, starch and sucrose metabolism, histidine metabolism, pyruvate metabolism, phenylalanine, tyrosine and tryptophan biosynthesis. The established metabolomic method can comprehensively evaluate the toxic effects of chlorophenol disinfectants on aquatic organisms, and provides theoretical basis for further elucidating the toxicity mechanism of chlorophenol disinfectants.