基于“体内过程-靶细胞结合”和“在线活性分析”的泻心汤抗动脉粥样硬化效应物质基础研究

Xie-xin Decoction has a significant effect for atherosclerosis in clinic. However, its active ingredients responsible for the beneficial effect to atherosclerosis remains unknown. Which has become the major obstacle for the clinical application and further pharmaceutical research of Xie-xin Decoction. This study will apply the rapid and convenient methods based on the combination of modern analytical techniques and pharmacological study to investigate the active ingredients of Xie-xin Decoction against atherosclerosis. Considering the synergistic effect of traditional Chinese medicine which are multi-component and multi-target in different pharmacological processes, “active ingredients online-screening”, “serum pharmaceutical chemistry” and “processes in vivo - target cell binding” will be used to study the correlations between chemical components and the main pharmacological processes, such as the protection of vascular endothelial cells, inhibiting proliferation of vascular smooth muscle cell, preventing macrophage-to-foam cell transformation, anti-inflammatory and antioxidant activity. From our study, it will preliminary clarify the active ingredients of Xie-xin Decoction against atherosclerosis, and the correlations between chemical components and the main pharmacological process. This study will provide a basis for clinical application and further research of Xie-xin Decoction. It will also provide a support for finding active ingredients and formula optimization of the traditional medicine.

泻心汤临床用于热毒型动脉粥样硬化具有显著疗效，然而其效应物质基础不明确，成为阻碍其应用和进一步研究开发的瓶颈。本项目基于现代分析技术和药理评价相结合的研究思路，采用符合复方作用特点且快速简便的方法，研究泻心汤抗动脉粥样硬化的效应物质基础。针对泻心汤抗动脉粥样硬化的主要药理作用环节和中药复方多组分、多靶点的作用特点，拟采用“在线活性测定”、“血清药物化学”和“体内成分-靶细胞结合”等技术相结合，从保护血管内皮细胞、抑制血管平滑肌细胞增殖、抑制巨噬细胞泡沫化、抗炎、抗氧化等防治动脉粥样硬化的主要药理作用环节，探讨泻心汤化学成分与药效的相关性。综合分析初步阐明泻心汤抗动脉粥样硬化的效应物质基础，解释特征化学成分与具体药理作用环节之间的关系。通过本项目，为泻心汤的临床应用和进一步研究提供依据，也为中药复方效应物质基础研究、处方解析与优化提供借鉴。

本项目采用HPLC-Q-TOF-MS/MS对泻心汤全方的化学成分进行定性分析，初步鉴定出40个化学成分，基本明确了泻心汤的化学组成；采用HPLC-ABTS-DAD-Q-TOF/MS联用在线分析泻心汤抗氧化活性成分，共分析鉴定出15个具有抗氧化活性的成分，初步阐明了泻心汤的抗氧化效应物质基础；采用血清药物化学的研究方法对泻心汤活性成分进行分析，共鉴定出21个入血成分，为阐明泻心汤的效应物质基础提供了依据；采用“靶细胞结合-液/质联用”技术，筛选泻心汤中与内皮细胞、平滑肌细胞和巨噬细胞结合的成分，最终确定9个化合物作为泻心汤有效成分组合；建立内皮细胞损伤模型、血管平滑肌细胞异常增殖模型、巨噬细胞泡沫化模型及斑马鱼动脉粥样硬化模型，在保护内皮细胞、抑制平滑肌细胞异常增殖、抑制巨噬细胞泡沫化、调节脂质水平、抗氧化等抗动脉粥样硬化主要药理作用环节比较泻心汤有效成分组合与全方的效应，从细胞水平和动物水平对泻心汤有效成分组合进行药效学评价，结果发现泻心汤有效成分组合在以上抗动脉粥样硬化关键药理作用环节的效应强度达到全方的80%以上，较好的代表全方发挥抗AS的作用，可认为是泻心汤抗AS的效应物质基础。