药物性肾损伤原位早期监测近红外纳米探针构建及其荧光可视化研究

Kidney is an important excretory organ of the human body and is often damaged by the toxic effect of drug or its metabolic product in the process of filtration, reabsorption and excretion. At the early stage, the process of the drug-induced renal injury is reversible. The renal function will be recovered if the injury is discovered and intervened at early stage, or the injury will be irreversible. Therefore, the establishment of early monitoring technology for the drug-induced renal injury is of great importance for the prevention of drug toxic and side effects and the improvement of success rate of new drug research. To overcome the drawbacks of low sensitivity, weak reliability and traumatic injury of the current detection methods, this research will develop new strategy to monitor the drug-induced renal injury at early stage which is based on fluorescent probe and imaging technology. This research will construct fast response near-infrared fluorescent nanoprobe to detect the ROS increase, hypoxia, pH change and calcium overload as the marker events at the early stage. The development of noninvasive, in situ and visualized in vivo imaging monitoring technology will increase the sensitivity, specificity and reliability of the methods. The research will also evaluate the performance of nanoprobe in living cells and in vivo, and optimize the working conditions and imaging parameters to establish the quantitative relation of “fluorescent properties of nanoprobe-occurrence degree of marker events-classification of drug-induced renal injury”, which will provide new technologies and methods for preclinical drug toxicity prediction and early diagnosis and treatment of drug-induced renal injury.

肾脏是人体重要的排泄器官，药物或其代谢产物在滤过、重吸收、排泄过程中容易引发毒性反应，造成药物性肾损伤。其过程早期为可逆反应，若能早期发现及干预，肾脏功能可逐渐恢复，否则易导致不可逆损伤，因此建立药物性肾损伤早期监测技术对避免药物毒副作用、提高新药研发成功率至关重要。针对当前肾损伤检测方法存在的灵敏度低、可靠性差、创伤性等问题，本项目拟基于荧光探针成像技术发展药物性肾损伤早期监测新策略：以药物性肾损伤早期标志性事件—ROS升高、乏氧、pH改变、钙稳态失衡为切入点，构建快速响应近红外纳米荧光探针，发展无侵害活体原位成像可视化监测技术，从而提高方法灵敏度、特异性及可靠性。在细胞及活体水平评价纳米探针应用性能，优化纳米探针最佳使用条件及成像参数，建立“探针荧光性质-事件发生程度-药物性肾损伤分级”三者之间的量化关系，为临床前药物毒性预测、药物性肾损伤早期诊断与治疗提供新技术与新方法。

肾脏作为机体不可或缺的排泄器官，在体内执行清除废物，维持电解质、酸碱平衡以及调节内分泌等一系列生理功能。由于大多数药物及其代谢产物通过肾脏排出体外，且肾脏血流丰富、代谢活性高，因此极易诱发药物性肾损伤（Drug-induced kidney injury，DKI）。随着药物种类的增加，新药的不断涌现以及部分中草药的不当使用，DKI的发生率逐年增加，已成为药物研发领域关注的焦点。本研究围绕肾损伤相关的活性物种与分子事件发展了一系列早期诊断新方法、功能成像探针与诊疗一体化体系，重点以DKI发生发展过程中细胞产生的活性分子（活性氧、酶）、生物标志物（NGAL、miRNA、CysC、MMP）、整体尿蛋白为分析对象，构建了若干种集肾细胞损伤早期检测、发展进程监测、肾损伤类型识别及中药保护作用评价的分子、纳米探针及荧光阵列传感器。阐明了分子探针与生物靶标的响应机理，确证了新方法相较于传统血肌酐、尿素氮检测方法的优越性，实现了肾损伤的快速体内外检测；基于荧光阵列传感技术建立了抗肿瘤药、氨基糖苷类抗生素、质子泵抑制剂以及非甾体抗炎药等临床50余种常见肾毒性药物的细胞损伤指纹图谱，为药物肾毒性评价与机制研究提供了快速分析方法；基于多标志物体外检测体系评价了中药炮制过程对何首乌的减毒效应、黄葵胶囊对肾间质纤维化的调控作用。此外，基于分子探针和脂质体技术建立了心肌乏氧成像检测平台，以功能探针获取了心肌损伤过程中硝基还原酶等生物活性分子的时空变化信息，研究了中药制剂生脉散对心肌乏氧的改善作用，为中医药现代化研究提供了高效分析工具及有益信息。利用青蒿素放大氧化损伤策略与分子探针技术构建了肿瘤精准诊疗一体化系统，为疾病早期诊断与高效治疗提供了新技术和新方法。本项目发表高质量研究论文20篇，申请中国发明专利16件，授权中国发明专利1件，培养博士研究生2名，硕士研究生10名。