百草枯中毒后代谢特征变化及生物标志性物的筛选

The high rate of paraquat poisoning induced-death, which is more dangerous than organophosphate pesticides, has become a social problem. However, the body's ability to detoxify after paraquat poisoning is unkown, which affects not only the understanding of paraquat toxicity, but also the design of clinical treatment after paraquat poisoning. It has become one of the urgent need to be solved clinical problems. This project intends to use metabolomics method, modern spectral analysis techniques, and cocktail assay to assess drug-related metabolic enzymes’ activity, combined the way of filtering biomarkers from the blood, urine and other biological samples ingredient after paraquat poisoning, and the relation between these biomarkers and intake, duration of action of paraquat to establish metabolomics mode in rats. Combined with pathological analysis, to establish the detoxification of the model based on this metabolomics to explore changes of metabolic capacity in rats after paraquat poisoning. Providing new ideas of accurate medication for clinical and forensic identification of clinical poisoning reference for paraquat toxicology, and this will also help in promoting the development of toxicology and emergency work after poisoning.

百草枯中毒致死率极高，它的危害性超过有机磷农药，已成为一项社会问题。但人们对百草枯中毒后机体解毒能力的改变规律还不了解，这不仅影响到对百草枯毒理的认识，也关系到百草枯中毒后临床救治方案的设计，已成为临床迫切需要解决的难题之一。本项目拟借用代谢组学的思路，现代波谱分析技术，及Cocktail法检测与药物相关代谢酶活力的改变的方法；并从血液、尿液和其他生物样品中筛选百草枯中毒的标志成分，及这些生物标志物与百草枯摄入量、百草枯作用时间的关系，建立百草枯中毒大鼠的代谢组学模式。同时结合病理分析，建立基于此代谢组学的大鼠百草枯中毒后机体解毒能力的模式，探讨大鼠百草枯中毒后代谢能力变化规律。为临床准确用药与法医临床毒化鉴定提供参考依据，为百草枯的毒理研究提供新的思路，同时，也有助于推动我国毒理研究和中毒后急救工作的发展。

本研究建立了采用梯度洗脱液相色谱-串联质谱法检测(UPLC-MS/MS)法对血浆探针药物非那西丁、甲苯磺丁脲、美托洛尔、咪达唑仑、奥美拉唑和安非他酮浓度进行测定，可以用到大鼠中毒模型中评价大鼠P450酶的活力。在这项研究中，我们开发了一个基于GC/MS的血清代谢组学方法评价急性百草枯中毒对大鼠的影响。经过模式识别分析，包括主成分分析和偏最小二乘法-判别分析显示急性百草枯中毒诱导代谢絮乱，与对照组相比硬脂酸，L-丝氨酸，苏氨酸，L-缬氨酸，和甘油在急性百草枯中毒组(36 mg/kg)增加，而棕榈酸，D-半乳糖，癸酸下降。这些发现提供了一个系统的概述对百草枯染毒和代谢组学了解百草枯的毒性机制。我们也开发了一个尿液代谢组学方法，基于GC/MS探讨急性百草枯中毒对大鼠的影响。经过模式识别分析，包括偏最小二乘法-判别分析和主成分分析表明急性百草枯中毒诱导代谢絮乱。与对照组相比，急性百草枯中毒组(36 mg/kg)中棕榈酸增加，而琥珀酸，戊二酸，阿卓糖酸的水平下降。基于这些尿代谢组学数据，支持向量机（SVM）应用于识别代谢组学变化百草枯组与对照组分类精度比较高。总之，代谢组学方法与支持向量机相结合可以作为百草枯中毒一个有用的诊断工具。