基于代谢组学方法研究通塞脉微丸治疗缺血性中风的机制

项目名称： 基于代谢组学方法研究通塞脉微丸治疗缺血性中风的机制

项目编号： No.30873401

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 文红梅

作者单位： 南京中医药大学

项目金额： 30万元

中文摘要： 本课题以通塞脉微丸为对象，结合色质联用和代谢组学技术，从整体观角度研究通塞脉微丸治疗缺血性脑中风的作用。 ①#24314;立通塞脉微丸色谱指纹图谱，标示出38个共有峰；建立通塞脉微丸多成分测定方法；以LC/ QTOF MS技术分析鉴定出其吸收入血成分共23个，整体控制通塞脉微丸质量。 ②#22312;此基础上，建立电凝中动脉致大鼠缺血性中风模型和线栓法复制大鼠中动脉栓塞模型，从病理指标、生化指标和代谢组学方法三方面对缺血性中风模型进行系统性研究，伴随脑缺血缺氧的发生，确定代谢组中异常变化的潜在生物标记物(电凝法27个，线栓法15个)涉及能量代谢障碍、脂质代谢和糖代谢紊乱、氨基酸代谢异常等，均与缺血性中风发生密切相关。 ③#37319;集假手术组、模型组、通塞脉组和阳性药组等动物中与缺血性中风相应的体液和组织（血浆、脑组织和脑脊液）进行代谢组学研究，发现通塞脉微丸可使造模后大鼠血浆及脑组织中异常变化的代谢物水平有不同程度的恢复，而造模前后大鼠脑脊液的成分相对稳定。 研究结果阐明通塞脉微丸治疗缺血性中风的作用通过对脂质代谢、糖代谢和氨基酸代谢等异常代谢途径的整体调节而发挥作用，为中药的药效作用整体评价及作用机制的探讨提供新方法。

中文关键词： 通塞脉微丸；质量；缺血性中风；代谢组学；作用机制

英文摘要： Ischemic Cerebral Apoplexy (ICA) is one of the most popular diseases in elder people around the world which takes severely damages to humans' health. Tongsaimai pellet is a traditional Chinese medicine preparation which was commonly used for treat ICA. In this study, the therapeutic effect of Tongsaimai pellet was studied by chromatography/mass spectrometry technology and metabonomics method. First, the quality of Tongsaimai Pellet was researched. Chromatographic fingerprints of Tongsaimai pellet were established and 38 common peaks were marked by UPLC method. Multiple components in Tongsaimai pellet were determined simultaneously by HPLC and UPLC. Using LC/QTOF MS technology, 23 components were identified in rat serum. Secondly, two rat models of ICA caused by electric coagulation on middle cerebral artery and model of middle cerebral artery occlusion with Suture Method were evaluated though metabonomics method comparing with pathological parameters and biochemical indicators. During cerebral ischemia and anoxia, potential biomarkers (27 in Electrocoagulation Method, and 15 in Suture Method ) with abnormal changes in metabolites were related to energy metabolism failure, lipid and glucose metabolism disorder, and amino acid metabolism disorder, which were all closely related to ischemic cerebral apoplexy. Thirdly, endogenous metabolits of bio-fluids (blood, brain tissue and cerebrospinal fluid) from sham-operation group，model group, Tongsaimai pellet group and positive control group in rats were determined with GC-MS using metabonomics method. Changed metabolities in blood and brain tissue could be certainly recovered by Tongsaimai pellets, while cerebrospinal fluid composition was relatively stable after modeling. Tongsaimai pellets can treat ICA by overall regulation of abnormal metabolism, such as lipid metabolism, glucose metabolism, amino acid metabolism, etc. These complementary relations can explain the mechanism of Tongsaimai pellets to the ICA modeled animal in an entireness way and provided a new pathway and methodology for pharmacological effects evaluation and mechanism research of TCMs.

英文关键词： Tongsaimai pellet; Quality; Ischemic Cerebral Apoplyexy; Metabonomics; mechanism of action