基于LasR受体在线筛选中药黄芩中细菌生物膜抑制活性成分的研究

项目名称： 基于LasR受体在线筛选中药黄芩中细菌生物膜抑制活性成分的研究

项目编号： No.81503031

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 郭嘉亮

作者单位： 暨南大学

项目金额： 17.9万元

中文摘要： 细菌生物膜（BF）是导致耐药行为的重要原因。群体感应抑制剂（QSI）通过拮抗细菌自身的群感系统而抑制BF的形成进而避免耐药性产生，是研发新型抗菌药物的重要突破口。中药是获得QSI的重要来源，具有低毒价廉、不易引起耐药等优势。然而，现有的筛选策略和方法存在着耗时低效、费用高昂等不足，严重制约了从中药复杂成分中发现QSI。前期，我们发现中药黄芩可以抑制铜绿假单胞菌（PA）的BF形成。本研究拟首次基于PA群感系统中与BF形成密切相关的重要受体蛋白LasR，建立LasR亲和整体色谱­纳流液相色谱­质谱二维在线筛选方法，寻找中药黄芩中具有抑制BF活性的新型QSI，并通过体外BF抑制活性试验验证，结合QSI选择器筛选法和计算机辅助手段进行相互验证。本项目旨在提供一种“筛选、分离、鉴别”一体化的中药QSI研究方法，为揭示中药抗耐药菌的科学内涵，开发安全、有效的抗耐药菌中药新药提供科学依据。

中文关键词： 群体感应抑制剂；亲和整体色谱；细菌生物膜；LasR；受体；黄芩

英文摘要： The formation of biofilm (BF) is one of important reasons for bacterial resistance behavior. Quorum sensing inhibitors (QSIs), which could interfere with the quorum sensing system and therefore inhibit BF formation without causing resistance, represent an important breakthrough in the development of new antibacterial drugs. Traditional Chinese medicine (TCM) as an important potential source of QSIs, exhibits some advantages such as low toxicity, low cost, and low tendency to induce resistance. However, current screening strategies and methods have some shortcomings, such as time consumption, low efficiency, high cost, etc., which seriously restricts the possibility of obtaining novel QSIs from TCM. In a previous study, a well-known TCM, Scutellaria baicalensis Georgi, has proved to inhibit BF formation. In this research, the QS receptor protein LasR, which is closely related to Pseudomonas aeruginosa BF formation, will be immobilized onto a monolithic column for the first time. In order to screen BF inhibitors from Scutellaria baicalensis Georgi extracts, a platform, which combines LasR-affinity monolith chromatography (LasR-AMC), RP-HPLC, and mass spectrometry (MS) technologies, will be established. The BF inhibition activity of the screened active ingredients will be verified by means of an in vitro activity test. In addition, a comparative analysis between AMC and other screening methods, such as QSI selectors and computer-aided drug design (CADD), will be carried out. This research will be useful to elucidate the scientific connotation of TCM in the fight against drug-resistant bacteria. Moreover, it can not only provide an effective screening tool, but also a significant basis for the development of novel, safe, and effective TCM drugs against resistant bacteria.

英文关键词： Quorum sensing inhibitor ;Affinity monolithic chromatography;Bacterial biofilm;LasR receptor;Scutellaria baicalensis Georgi