磺酰氯类物质衍生化的BPs-EM-NT定量靶向代谢组学UHPLC-MS/MS平台构建及其应用

Research on the effects of environmental pollutants on metabolic pathway by metabonomics is one of the hot spots in the field of public health. Bisphenol A (BPA) is a recognized environmental endocrine estrogen disruptor, while recent studies showed that BPA analogues, BPs, also played a similar role, with quasi or anti estrogen effects. BPs may interfere with the metabolic balance and transformation of estrogen metabolites (EM) and neurotransmitters (NT), which may be closely associated with a variety of estrogen-related diseases and nervous system diseases. Thus, the present study is aimed to establish a high-throughput and high sensitivity targeted metabolomics UHPLC-MS/MS platform to quantify sulfonyl chloride-derived BPs-EM-NT substances. The derivatization reagents and conditions will be optimized by screening sulfonyl chloride compounds, and then BPs, EM and NT from biological samples will be derivatized, and the processing methods on the detection of the free and total amount of these substances in biological samples will be evaluated so as to optimized chromatographic mass spectrometry conditions. The biological samples will come from patients with depression, with the assay aforementioned, we will measure and evaluate the three types of substances to explore the interference mechanism of BPs in estrogen and neurotransmitter metabolism. Thererfore, the platform will provide important avenues and supports for the BPs exposure monitoring, the possible interactions between estrogens and neurotransmitters, and neurological or endocrine system-related diseases.

应用代谢组学方法研究环境污染物对机体代谢通路的影响是公共卫生领域研究的新热点之一。双酚A（BPA）是公认的环境类雌激素干扰物，研究发现其类似物BPs作为替代物也能发挥类似BPA的拟/抗雌激素作用。BPs进入机体干扰体内雌激素（EM）和神经递质（NT）的代谢平衡和转化，这可能与多种雌激素相关疾病和神经系统疾病密切相关。本课题拟建立高通量高灵敏度的磺酰氯类物质衍生化的BPs-EM-NT定量靶向代谢组学UHPLC-MS/MS平台。通过筛选磺酰氯类物质确定BPs、EM和NT最佳衍生化试剂及条件，比较研究三类物质总量与游离态的样品前处理方法（SPE和LLME），优化色谱质谱测定条件，进行方法学评价。收集抑郁症患者生物样本，用以上平台对三类物质进行测定与评价，寻找BPs在抑郁病中对雌激素和神经递质代谢的干扰机制。该平台的建立可为BPs暴露监测及对机体的影响、雌激素和神经递质交互作用等研究提供基础支持。

近年来双酚类物质（BPs）的环境毒性引起人们的重视。BPs对神经系统的扰乱作用和机制还未见深入研究。本项目建立了高通量、高灵敏度的磺酰氯类物质衍生化的神经递质（NT）、雌激素（EM）和双酚类物质（BPs）定量靶向代谢组学UHPLC-MS/MS平台。通过结构探究及质谱解析确定NT、EM和BPs丹磺酰氯衍生化产物的母离子及子离子；通过筛选磺酰氯类衍生化试剂确定了最佳衍生化试剂为丹磺酰氯，确定了衍生化反应发生的最佳pH值、温度、时间和浓度；建立了血清、脑组织、尿液、肠组织中神经递质UHPLC-MS-MS靶向代谢组学检测平台，并应用此平台研究了BPs（包括BPA、BPS、BPF）暴露亚慢性染毒模型小鼠神经递质代谢平衡的变化情况；研究了围产期暴露BPA引起子代小鼠神经递质代谢平衡紊乱机制；研究了抑郁样小鼠不同脑区及不同肠段神经递质的分布与表达情况；研究了BPF在非酒精性脂肪肝样改变中的作用。因此，本项目中定量靶向代谢组学BPs-EM-NT的UHPLC-MS/MS检测平台的建立为深化BPs暴露监测和对机体的影响、雌激素和神经递质之间可能的交互作用以及神经、内分泌系统相关疾病等的研究提供了重要基础支持。