基于质谱的细胞代谢组学方法及其在新型有机污染物的毒性评估中的应用

The study on the risk assessment and human beings exposure of the emerging chemicals are the hotspot in the environmental research in recent years．In vitro cytotoxicity evaluation by induced pluripotent stem cells, as an alternative of traditional toxicity evaluation using animal models, has been widely studied and applied.　AS an emerging filed, cell metabonomics is defined as the set of small molecule metabolites in intracellular and reflect the response to enviromental change. and it has become a promising analytical tools for toxicity assessment. In this study, a sensitive method to analysis the intracellular metabolites using UPLC/QTrap MS, UPLC/QTof MS and UPLC/QQQ MS will be established; Emerging organic pollutants will be exposed to induced pluripotent stem cells, whose exposure dose equals to the dose in enviroment,which will be used to evaluate the cell metabolites disorders.Integration of data processing methods, bioinformatics and cell toxicity will be adopted for studying the pathway and toxic mechanism of pollutants, which will provide data support for environmental control and risk assessment system．

新型有机污染物的研究是目前国际上环境化学研究的热点领域，因其环境污染引起的人体健康问题是我国环境风险评估的重要组成部分。体外细胞模型诱导多能干细胞作为传统动物模型毒性评价的替代方法正在得到越来越多的研究和应用。细胞代谢组通过定性、定量地描述胞内调控的最终产物，表现生物系统对环境改变的最终应答，已成为一个有前途的毒理学评价的分析工具。本研究拟采用多种串联质谱技术相结合，建立一套适用于细胞内代谢物检测的高通量、高灵敏分析方法，选择人体诱导多能干细胞为暴露模型，将等同于环境剂量的新型污染物的标准品暴露于细胞中，探索新型污染物对细胞代谢物的干扰作用；结合统计学、生物信息学及细胞毒性实验，定位污染物影响的代谢路径，揭示毒性作用机制，为污染物的风险识别和实施污染控制提供数据支持。

氯代多环芳烃Cl-PAHs已成为一类新型的高风险有机污染物，对生态环境和人类健康构成了潜在威胁。然而其的毒性效应及代谢毒理学研究十分有限，限制了对其进行生态风险和人体健康风险评估。本项目建立了“全靶向”细胞代谢组分析方法与最大限度地提取极性以及非极性代谢物的细胞样品前处理方法，以人体正常肝细胞LO2为模式细胞，基于发展的"全靶向"分析代谢组学研究策略，研究了不同Cl-PAHs单体暴露所引起的生物代谢改变。结果表明即使环境剂量的Cl-PAHs也会引起细胞代谢的显著改变（以代谢影响水平指数为评价指标）;Cl-PAHs暴露可引起LO2细胞活性降低; Cl-PAHs暴露对代谢通路的干扰与其母体PAHs并非完全一致。