具有靶向功能松香基比率荧光探针的设计合成及对细胞色素P450亚类2J2检测性能研究

At present, there are still many problems about the fluorescence detection probes for cytochrome P450 2J2, such as selectivity, sensitivity and single function. These hardly meet the requirements of practical applications. This project plans to design and prepare a bifunctional ratiometric fluorescent nanoprobe containing nucleic acid aptamers, rosin monomers, hydrophobic carbon dots, and functional fluorescent dye molecules by means of utilizing living radical polymerization, nano-precipitation technology and functionalization technology of aptamers. This fluorescent probe will detect the endogenous CYP2J2 in acute lymphatic leukemia cells based on fluorescence resonance energy transfer (FRET). Combined with UV-Vis and FS, the fluorescence responses of this probe upon exposure to CYP2J2 with different ratios of donor and receptor will be investigated. The association between spectral parameters and the system structure wil be confirmed. The mechanism of this ratiometric sensing system will be elucidated. The targeted mechanism will be illustrated by investigating the cytotoxicity, cellular intake level and fluorescence imaging of probes. We will explore the feasibility of determining the existence state of CYP2J2 in acute lymphatic leukemia cells. This study will provide the new evidence for the design and synthesis of rosin-based fluorescent probes, and basis for the research of the biological function of CYP2J2 in acute lymphoblastic leukemia cells.

目前CYP2J2荧光探针在选择性、灵敏性方面的局限性，且功能单一化，无法满足应用要求。本项目拟通过活性自由基聚合、纳米沉淀技术及适配体修饰技术等手段，将核酸适配体、松香单体、疏水性碳点和高选择性识别能力的功能化荧光染料单体有机结合，构筑双功能荧光纳米探针，利用荧光共振能量转移原理，实现对急性淋巴白血病细胞内CYP2J2高选择性、高灵敏度比率检测。综合各种手段优化体系中供受体的比例和能量转移行为，结合紫外-可见吸收光谱、荧光光谱等方法研究不同供受体比例下的荧光响应特征，确定光谱特性参数与体系结构间的构效关系；阐明松香基聚合物的CYP2J2荧光比率检测机制；考察探针的细胞毒性、细胞摄入水平和荧光成像，探索急性淋巴白血病细胞内CYP2J2的水平和时空分布，揭示探针对肿瘤细胞的靶向识别规律，将为松香基荧光探针的设计合成方面提供有益参考，也可为急性淋巴白血病细胞中CYP2J2的生物功能提供新的依据。

生物分子、离子等物质含量在生命体系和环境保护方面发挥着重要的作用，对它们的精准检测对疾病早期诊断和污染物识别具有重要意义。本项目以官能团修饰、薄膜水化组装、表面修饰、溶剂热等策略，构建一系列具有生物活性分子、离子特异性识别的荧光（纳米）探针和电化学发光传感器，系统深入地探索了荧光探针中反应活性基团与生物活性分子、离子之间的反应关系，荧光材料配比及浓度与荧光强度变化的关系以及能量转移效率优化等问题，并将它们用于水/细胞中生物活性分子、离子的高性能荧光/电化学发光检测和生物成像。本项目所取得的成果为光学材料的设计合成与生物成像、电化学发光传感应用研究等方面提供新思路。从项目立项至今，已先后在Dyes and Pigments、Microchemical Journal、Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy、Analytical and Bioanalytical Chemistry、Chemosensors、分析化学等国际国内学术期刊发表相关工作6篇学术论文。申请国家发明专利2件，并且有望在环境监测、医疗诊断和光学农业等进行实际应用。