基于信号靶点识别的敏感细胞传感检测呕吐毒素及其降解产物毒性评价体系的建构研究

Currently, the evaluation of mycotoxins degradation technology is on the basis of the reduction of mycotoxins, however, there is few researches on the potential toxicity of the degradation products, it is hard to make correct evaluation on food safety after mycotoxins degradation. The project is based on the deoxynivalenol（DON）target signaling pathways to build new cell sensing model with specific recognition of DON and its degradation products; The effective disciplines of toxicity pathway signal transduction, receptor-ligand interactions, fluorescence signal conversion and cytokines induced expression were developed before and after degradation; Combined with high content screening and electrochemical sensor, a variety of degradation products of multi-target detection parameters was obtained, including the corresponding variation characteristic rules; Based on the cytomics, screening and validation are used to establish the fast accurate evaluation of the potential DON degradation products toxicity system. The results establish a theoretical foundation for the variation of the poisonousness on different degradation conditions for the mycotoxins, which can be applied to further guidance and correct the degradation method, achieve active control of food security and provide a necessary protection during food production, storage and transportation and provide theoretical support for the realization of intelligent in situ, real time, and early monitoring of food chain.

目前评价粮食中真菌毒素消减效果主要是以毒素含量降低为依据，而对降解过程中产物的潜在毒性的监测缺乏足够研究，难以科学评价毒素消减技术应用的安全性。本项目基于呕吐毒素毒性信号通路靶点，构建快速、可直观监测且能够特异性识别呕吐毒素及其降解衍生物毒性作用的新型细胞传感评价模型；研究毒素降解前后对细胞信号通路受体识别、荧光信号转化以及细胞因子诱导表达的影响规律；通过毒性通路中细胞受体-配体相互作用，建立呕吐毒素降解产物与敏感细胞“剂量-时间-效应”关系，结合高内涵筛选和电化学传感，实现对毒素降解产物多靶点参数的同时检测，解析细胞特征信号的变化规律；基于细胞组学，对细胞筛查模型进行修正和验证，建立快速准确评价呕吐毒素及其降解产物细胞潜在毒性的模型体系。研究成果应用于指导和修正毒素消减模式，为粮食的安全生产、储存和加工提供必要保障，对实现食品链中潜在毒素原位、实时、早期监测和预警奠定了坚实的理论基础。

针对粮食加工中真菌毒素消减过程，研究特异性识别DON信号通路关键靶点AP-1 site，以特异性结合位点为毒性通路标志物，构建基于AP-1 site-mCherry信号级联序列的快速、可直观监测，特异性识别DON及其降解产物毒性的新型细胞传感评价新方法；结合纳米介导细胞传感，建立毒素降解产物与敏感细胞“剂量-时间-效应”关系，追踪毒性物质产生的生物学信号；结合转录组和代谢组，研究消减产物对细胞靶向作用的关键靶点和作用机制，发现毒素暴露导致SIRT1、SOD上调引起的细胞氧化应激；通过组学技术，对DON及其降解产物联合诱导的细胞毒性效应研究，发现臭氧降解影响甘氨酸、丝氨酸、苏氨酸代谢、亚油酸代谢，氰氨基酸代谢等多条代谢通路；进一步解析臭氧降解条件下DON降解产物毒性变化规律，建立臭氧降解DON过程的动力学模型，鉴定毒性相关的中间产物，解析降解路径，指导和修正毒素消减模式。研究结果可为粮食安全生产、储存和加工提供必要保障。