以TolC蛋白为靶标的大肠杆菌外排泵抑制剂的筛选及耐药干预机制研究

项目名称： 以TolC蛋白为靶标的大肠杆菌外排泵抑制剂的筛选及耐药干预机制研究

项目编号： No.81202451

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 药物学、药理学

项目作者： 陈伶利

作者单位： 湖南中医药大学

项目金额： 23万元

中文摘要： 多重药物外排泵广泛存在于各种病原菌中, 对细菌的药物敏感性以及细菌性感染的临床治疗影响很大，所以外排泵抑制剂是解决细菌耐药问题的重要途径。然而，目前还没有理想的应用于临床的抑制剂产生，找到适用于临床的外排泵抑制剂显得尤为重要。本研究以大肠杆菌为模式菌，以构成外排泵的重要组成成分－外膜蛋白TolC为靶目标，拟通过SELEX技术筛选一组关于TolC的高特异性和亲和性的适配体，利用适配体与靶蛋白结合后能抑制蛋白质活性的特点，开展适配体阻塞大肠杆菌多重外排通道以及抑制耐药的研究；同时结合MALDI-TOF/MS、SPR、纳米磁性粒子分离等技术对抑制剂与外排泵的相互作用机制进行探讨。鉴于适配体的优势和外排泵在多重耐药中的重要作用，此项研究将为外排泵抑制剂的研究提供有力支持，为我国解决细菌耐药性的问题开辟新的思路，同时为我国耐药性的临床干预提供一种特异性强、灵敏度高和易操作的、安全的技术和手段。

中文关键词： 大肠埃希菌；外膜蛋白tolC；外排泵抑制剂；适配体；SELEX

英文摘要： Multiple drug-resistant efflux pump is widely existed in all kinds of pathogens and has great influence on the drug sensitivity of bacteria and the clinical treatment of bacterial infection. Therefore, Efflux pump inhibitors (EPIs) is an important way to solve the problem of bacteria resistance. However, no ideal medicine has been clinically applied in the current researches of EPIs. As a result, it is particularly important to find a clinically EPIs. Escherichia coli is used as the model bacteria and targets to constitute the outer membrane protein TolC which is the major composition of the efflux pump in which plans to use the SELEX technology to get a group of aptamer with high specificity and compatibility and take advantage of the feature that the protein activity can be restrained with the combination of the aptamer and the target protein to study the Escherichia coli multiple efflux passageway blocked by the aptamer. It will be also discussed what is the functional mechanism of inhibitor combined with the technologies of bio-nano-materials, MALDI-TOF/MS and SPR. Considering the advantages of aptamer and importance of efflux pump in multiple drug resistance, this project will not only provide a type of direct and safe technology and method with high sensitivity and specificity to the clinical intervene of

英文关键词： E. coli；Out membrane protein tolC；Efflux pump inhibitor；Aptamer；SELEX analysis