基于代谢组学的冠心舒通胶囊防治冠心病的药效物质基础和作用机制研究

Guanxinshutong (GXST) capsules have a certain and positive therapeutic effect against coronary disease. However their therapeutic basis and the action mechanism are not very clear. Using the method of syndrome differentiation and treatment in Traditional Chinese Medicine (TCM), GXST capsules will be studied to evaluate the effects based on metabonomics research strategy, by using different coronary disease pathology animal models with pharmacological indexes and biochemistry measurement and combining the results of serum pharmacology research. The action mechanism of GXST capsules will be investigated by the method of pharmacokinetics and pharmacodynamics. The fingerprints of GXST capsules in vitro and vivo will be established with analysis techniques such as LC-MS, and the therapeutic basis will be characterized together with serum pharmacology, pharmacokinetics and pharmacodynamics. Meanwhile, the metabonomics studies of GXST capsules will be carried out. The biomarkers closely related with the action mechanism of GXST capsules will be found through analyzing the feature changes of the established metabolic profiles of serum and urine of different groups. In a word ,embarking on concept of the holism for TCM theories, the therapeutic basis, compatibility and action mechanism of GXST capsules will be illustrated by the way of combinations of in vitro and in vivo study, chemical data and pharmacological data, metabolomics data and pathological physiology along with biochemistry indexes. The research will provide a theoretical support for the clinical application and spread of GXST capsules, and will explore a new way with holism, efficiency and systematicness aiming at TCMs.

冠心舒通胶囊临床治疗冠心病疗效确切，但其药效物质基础和作用机制不甚明了。故本课题以冠心舒通胶囊为研究对象，以其功能主治为依据，基于代谢组学研究策略，构建冠心病病理大鼠动物模型，进行药理和生化指标检测，结合中药血清药理学结果，科学评价其防治冠心病的药效；同时考察药效物质药动学行为，药效-药动相结合，对冠心舒通胶囊进行药效动力学研究，阐述其作用机制；采用LC-MS等先进分析手段，建立冠心舒通胶囊的体外和体内指纹图谱，并与药物动力学和药效动力学相关联，确定其药效物质群；对冠心舒通胶囊进行代谢组学研究，针对有效的冠心病病理大鼠模型，建立并分析其血清和尿液的"代谢谱"，找寻与作用机理密切相关的生物标志物；对体内外指纹图谱、生化药理指标、药物动力学和药效动力学行为以及代谢组学的大量数据信息进行统计分析，系统评价其功效内涵，阐明其发挥防治冠心病药效的物质基础，揭示其作用机制,为其临床应用与推广提供依据。

本课题以冠心舒通胶囊为研究对象，以其功能主治为依据，基于代谢组学研究策略，构建了动脉粥样硬化、急性血瘀和急性血栓大鼠动物模型，同时进行药理和生化指标检测，结合中药血清药理学结果，表明了冠心舒通胶囊对冠心病发作不同阶段防治有明显作用。通过测定16批冠心舒通胶囊样品，建立了冠心舒通胶囊HPLC指纹图谱，并通过对照品对共有峰进行指认，标定了45个共有色谱峰，34个归属到各药材，考察了全方与各组成药材指纹图谱的相关性，同时采用HPLC法测定了其中13种主要化学成分的含量。建立了冠心舒通胶囊在大鼠体内的UFLC血清指纹图谱，采用UHPLC-Q-TOF-MS技术对冠心舒通胶囊的体外和体内化学成分进行分离鉴定，在胶囊中共检测到73个色谱峰，在健康大鼠血浆中共检测到44种入血化学成分：包括23种原形化合物和21种代谢物。建立了同时测定大鼠灌胃给予冠心舒通胶囊后血浆中8种相关成分的UPLC-MS/MS方法，并对其在健康大鼠和动脉粥样硬化症大鼠体内的药动学行为进行比较；采用UFLC-MS/MS技术，对冠心舒通胶囊中9种活性成分在健康大鼠和急性血瘀大鼠体内的药动学进行了对比研究，建立了同时测定大鼠血浆中 5种酚酸类和4种二萜类化学成分的超高效液相色谱-三重四极杆串联质谱法，并对冠心舒通胶囊全方和各缺味药材组方，分别进行了单复方药动学对比研究。建立了同时测定大鼠血浆中丁香酚等4种挥发性化学成分含量的GC-MS法，并对分别灌胃冠心舒通胶囊全方和单味药材后的大鼠体内药动学行为进行了对比研究。应用代谢组学方法对动脉粥样硬化和急性血瘀模型大鼠的血浆、尿液进行了分析，并对冠心舒通胶囊针对冠心病的的治疗及预防作用机制进行了探讨；采用主成分分析法对原始图谱数据进行分析，得到得分图和载荷图，并且分别在高血脂模型大鼠和急性血瘀模型大鼠的尿液及血浆中共鉴定出生物标志物。本课题基本完成了课题的预期目标，为冠心舒通胶囊的研究开发和临床应用提供了实验依据。