药物作用机理分子印迹生物传感器新方法的构建及对中药活性成分丹参酮ⅡA萃取-检测和活血化瘀功效药效学的研究

TanshinoneⅡA, as an active ingredient of traditional Chinese medicine Danshen, has been proven by numerous experiments that it has a positive impact in the treatment for cardiovascular diseases. However, some common testing methods are difficult to meet the strict requirements of detection for the active components of traditional Chinese medicine. Based on the rapid developments and abundant findings in the field of biosensing and traditional Chinese medicine, we propose to establish a comprehensive biosensing system for the effective components of single Chinese medicine. This project uses biosensing technology, innovatively combining molecular imprinting and solid phase extraction, to provide a new system which can be applied in extraction, detection and study in pharmacodynamics of traditional Chinese medicine TanshinoneⅡA. According to the main approaches and targets for platelet aggregation, three biosensing reseach systems will be constructed, namely an electrochemical biosensing research system for thrombus A2 pathway, an optical biosensing research system for ADP induced platelet aggregation, and a latter biological sensing system for the pathway of platelet glycoprotein GPⅡb/Ⅲa complex and its receptor. Overall, the project will present an integrated biosensing system to elaborate the multi-target and multi-channel mechanism of tanshinone ⅡA from the level of modern molecular biology, thus providing a new technology and new method for research of traditional Chinese medicine.

丹参酮ⅡA是中药丹参的活性成分，实验证明其具有改变血液流变性作用，可有效治疗心血管疾病,但用于其研究的专用技术方法尚需改进。在生物传感和中医药领域的研究成果指导下,本项目试图对中药有效成分建立完整的生物传感研究体系。本研究使用生物传感技术，结合分子印迹、固相萃取等方法，将多种生物传感技术应用于中药活性成分的萃取-检测及药效学的研究。结合新技术构建分子印迹生物传感体系实现对丹参酮ⅡA的特异性识别并检测。创新性地与固相萃取技术联用，构建出丹参酮ⅡA萃取-检测体系。针对血小板聚集的主要途径和靶点分别构建出针对血栓素A2途径的电化学生物传感研究体系、基于ADP诱导血小板聚集作用的光学生物传感研究体系以及根据血小板糖蛋白GPⅡb/Ⅲa复合物与受体结合途径的质量型生物传感研究体系。试图从现代分子生物学水平阐述丹参酮ⅡA多途径多靶点的药效机制，为中医药的研究提供新技术新方法。

丹参酮ⅡA作为中药丹参中的有效活性成分，因其具有改变血液流变性的特性，可有效治疗心血管疾病，但用于其研究的专用技术方法尚需改进。本项目在生物传感和中医药领域研究成果指导下，成功构建了完整的丹参酮ⅡA生物传感研究体系。结合分子印迹、固相萃取等方法，将多种生物传感技术应用于中药活性成分的萃取-检测及药效学研究。构建了分子印迹生物传感体系实现对丹参酮ⅡA的特异性识别及检测，具有灵敏度高、稳定性好、检测限低等优势。创新性地与固相萃取技术联用，构建了丹参酮ⅡA萃取-检测体系。针对血小板聚集的主要途径和靶点分别构建了针对血栓素A2途径的电化学生物传感研究体系、基于ADP诱导血小板聚集作用的光学生物传感研究体系以及根据血小板糖蛋白GPⅡb/Ⅲa复合物与受体结合途径的质量型生物传感研究体系。本项目从现代分子生物学水平阐述了丹参酮ⅡA多途径多靶点的药效机制，进一步推动生物传感技术在中医药研究中的应用。