肝细胞线粒体内双活性氧多通道探针研究

Reactive oxygen species hydrogen peroxide and peroxynitrite produced through oxygen metabolism of life are closely related to a variety of biological activity and disease. They are involved in different biological process and interact with each other. It is of great significance to find ideal probes to detect and discriminate these species for the study of the life process, diagnosis of diseases and drug screening. That produced in the liver tissue has become one of the important biomarkers for the assessment of drug-induced hepatotoxicity by the U.S. Food and Drug Administration. In this project, a platform of self-assembly is constructed, where amphiphilic polymers are used as carriers to capsulate hydrophobic excited state intramolecular proton transfer (ESIPT) molecules in their hydrophobic cavities. The platform has the capability of targeted delivery. Through the introduction of coumarin fluorophore that can turn-on response to hydrogen peroxide, the simultaneous detection and differentiation of hydrogen peroxide and peroxynitrite with multi channel response and signal amplification would be realized. The assembly targeted on mitochondria of hepatic cells is further used to detect the hydrogen peroxide and peroxynitrite induced by drugs including acetaminophen and isoniazid to monitor in vivo hepatotoxicity caused by drugs. The project will further broaden the application of the amphiphilic polymers and ESIPT molecules in the biological field, and provide new theoretical guidance and scientific basis for the simultaneous detection and differentiation of two kinds of biomolecules with multi channel response, study of life activities, disease diagnosis, and drug screening.

生命体有氧代谢过程中产生的活性氧物质过氧化氢和过氧亚硝基与多种生物活动和疾病密切相关，虽然它们参与不同的生物过程但相互作用，寻找能检测和区分这些物质的探针，对于生命过程研究、疾病诊断和药物筛选具有重要的指导意义。在肝组织产生的活性氧物质已成为美国食品药品监督管理局评估药物肝毒性的重要指标。本申请将两亲高分子作为载体，通过包裹疏水性激发态分子内质子转移（ESIPT）分子形成组装体。通过引入过氧化氢诱导的香豆素，可实现多通道、信号放大、同时检测和区分过氧化氢和过氧亚硝基。将该组装体靶向定位肝细胞线粒体，可进一步用于扑热息痛、异烟肼等药物诱导肝组织产生的过氧化氢和过氧亚硝基检测，评估药物的肝毒性。本项目的研究将拓宽两亲高分子和ESIPT分子在生物领域的应用范围，为多通道同时检测两种生物分子、生命活动研究、疾病诊断和药物筛选等提供理论指导和科学依据。

生命体有氧代谢过程中产生的活性氧物质过氧亚硝基与多种生物活动和疾病密切相关，虽然它们参与不同的生物过程但相互作用，寻找能检测和区分这些物质的探针，对于生命过程研究、疾病诊断和药物筛选具有重要的指导意义。在肝组织产生的活性氧物质已成为美国食品药品监督管理局评估药物肝毒性的重要指标。本项目合成了一系列激发态分子内质子转移（ESIPT）分子，并构建功能分子组装体，实现了多通道、信号放大、细胞定位、能同时检测和区分两种物质等功能。探索取代基对ESIPT 分子荧光及检测的影响，研究其构效关系，为设计更多性能良好的基于ESIPT 机理的荧光传感器用于两种物质同时检测提供理论指导；通过与不同取代的苯硼酸组装，实现了信号放大用于ONOO-的检测和细胞成像，药物肝毒性评估、药物筛选等提供前期的理论指导。