苯并三氮唑及其衍生物的分子作用位点甄选和作用机制研究

Benzotriazole and its derivatives (BTRs) have became an emerging contaminant for their high-production volume, widely detected in environments and potential adverse effects on organisms. At present, most toxicity studies on BTRs were based on the observation of impairments occurred on individual and organizational levels, which were detected after acute exposure or short-term and high-does exposure. Moreover, there are data gaps in regard to their endocrine disrupting effect at the molecular levels and the range of toxic mechanisms underlying BTRs exposures are not completely characterized. As a result, new evidences on molecular toxicity and toxic mechanisms are needed. In this research, zebrafish is chosed as model organism and will be exposed to BTRs for whole life cycle in low dose. After exposure, liver proteins will be analyzed by phosphoproteomics and the signal networks will be established. In addition, molecular targets will be screened using QPCR and western blot. Based on these, toxic effects and mechanisms on mRNA and protein levels will be preliminarily characterized. In combination of biochemical indices and histopathology observations, toxic mode of action and mechanisms are conducted to be investigated. This study will provide molecular toxicology for BTRs hazard prediction.

苯并三氮唑及其衍生物(benzotriazole and its derivatives, BTRs)由于生产量大，分布广泛以及对生物体具有潜在毒性成为一类新型污染物。目前对于BTRs的毒性研究多为急性毒性和短期高剂量暴露下的个体和组织水平上的表型指标变化，而且关于其内分泌干扰效应的作用模式仍不明确，需要在细胞和分子水平上开展BTRs的分子毒性效应和毒性机制研究。本课题以斑马鱼为模式生物，对其进行BTRs的低剂量长期暴露，以磷酸化蛋白组学技术为基础，构建BTRs对鱼类的毒性作用网络，结合基因表达和western blot免疫印迹技术甄选BTRs对鱼类的分子作用位点，从蛋白水平和基因水平上确定BTRs对鱼类的影响，初步阐明BTRs的分子毒理学机制。结合生化指标的测定和组织病理学观察，共同探讨BTRs的毒理作用模式和作用机制，为预测BTRs的危害性提供分子毒理学基础。

苯并三氮唑及其衍生物(benzotriazole and its derivatives, BTRs) 是一类具有优良的金属防腐蚀功能和光稳定功能的化学品，广泛应用于飞机防冻剂、塑料添加剂、护肤品等工业产品和个人护理品中，由于生产量大，分布广泛以及对生物体具有潜在毒性成为一类新型污染物。为了探明BTRs对鱼类的毒性效应和作用机制，研究首先以稀有鮈鲫为模式生物，用0.05, 0.5和5 mg/L BT对其进行28天暴露，以蛋白组学技术为基础，证明了BT可能通过干扰氧化应激、神经传导等生物学过程，对鱼类产生神经毒性和肝脏毒性；为了进一步研究其衍生物的毒性效应，选取典型苯并三氮唑紫外稳定剂（Benzotriazole ultraviolet stabilizers, BUVSs）UV-234，UV-326，UV-329和UV-P对斑马鱼幼鱼进行暴露，通过转录组测试发现，四种BUVSs均可影响幼鱼脑组织中与免疫通路相关的基因表达；最后，选取UV-234和UV-320两种典型BUVSs对斑马鱼胚胎进行暴露，发现二者的作用模式存在差异：UV-234通过抑制线粒体呼吸产生氧化应激从而导致斑马鱼胚胎延迟孵化，产生发育毒性；而UV-320则通过刺激线粒体呼吸导致大量活性氧生成，并间接作用于细胞炎症反应趋化因子cxcl-C1c和il8，产生免疫毒性。综上，BTRs可对鱼类的产生发育毒性和神经毒性，然而不同结构的化合物其作用机制存在差异，需要进一步研究。本项目研究为BTRs的毒性效应和作用机制提供了新的视角，为风险评价和化学品管理提供理论依据和技术支撑。