Corilagin干预动脉粥样硬化易损斑块作用及分子机制研究

To study the effect and mechanism of drug on intervening vulnerable plaque is of great significance for clinical preventing and curing acute coronary syndrome（ACS）. Our previous researches showed that Corilagin, an active ingredient from Yunnan featured medicinal plant Phyllanthus, had obvious effects against the experimental thrombosis and atherosclerosis and showed a potential effect on stabilizing vulnerable plaque. Based on these results, the atherosclerotic model of high fatty diet combined with balloon injury in miniature swine will be duplicated to evaluate the effects of Corilagin on atherosclerotic vulnerable plaques and its dose-effect relationships; in vitro, macrophages model are prepared and the intervening links of key genes involved in vulnerable plaque development will be investigated both in animal model and cell model; the modulation of Corilagin on CD40-CD40L signal pathway will be also determined in atherosclerotic vulnerable plaques and the cultured cells through the detection of key signaling molecules. The interventional effects on atherosclerotic vulnerable plaques of Corilagin will be explored systematically at these different levels such as model animals, vulnerable plaques, cells and molecules, in order to provide more scientific bases and experimental evidences for curing ACS with Corilagin.

研究药物干预易损斑块的作用及机制对临床防治急性冠脉综合征有重要意义。项目组前期研究发现，源于云南特色药用植物叶下珠的活性成分-Corilagin具有良好的抗血栓和抗动脉粥样硬化作用，并显示出潜在的稳定斑块作用。在此基础上，拟复制小型猪易损斑块模型，研究不同剂量的Corilagin治疗易损斑块的药理作用及其剂量-效应关系；在整体动物和体外培养的巨噬细胞模型上，寻找干预易损斑块形成发展的关键基因及其相互关系，研究Corilagin对CD40-CD40L信号通路关键因子的调控作用机制。以期从整体动物、易损斑块组织、细胞和分子水平系统研究Corilagin治疗易损斑块的作用，探索其细胞分子生物学机制，为将Corilagin研究成为自主创新和云南特色植物来源的治疗急性冠脉综合征药物提供进一步的理论和实验依据。

项目组前期研究发现，源于云南特色药用植物叶下珠的活性成分-Corilagin具有良好的抗血栓和抗动脉粥样硬化作用，并显示出潜在的稳定斑块作用。在此基础上，拟复制小型猪易损斑块模型，研究不同剂量的Corilagin治疗易损斑块的药理作用及其剂量-效应关系；在整体动物和体外培养的巨噬细胞模型上，寻找干预易损斑块形成发展的关键基因及其相互关系，研究Corilagin对CD40-CD40L信号通路关键因子的调控作用机制。结果表明，在整体动物模型上，Corilagin对小型猪实验性AS有明显的治疗作用，可降低小型猪血清中TC、TG、LDL-C、Apo B、Lp（a）水平，升高血清中HDL-C、Apo E水平； Corilagin能减少易损斑块脂质沉积和炎症反应，减轻动脉血管的病变程度、抑制胶原纤维的增生。Corilagin可降低小型猪AS模型颈动脉中膜厚度，减少易损斑块的数量，降低易损斑块面积，增加易损斑块稳定性，降低易损斑块中巨噬细胞的数量。Corilagin能明显下调AS斑块中TNF-α、NF-κB、MMP-1, 2, 9蛋白质和mRNA的表达。在细胞模型上，Corilagin能抑制巨噬细胞增殖，可将巨噬细胞阻滞于S期从而抑制其有丝分裂；Corilagin可明显减轻巨噬细胞泡沫化程度，降低巨噬细胞内脂质含量；Corilagin可抑制巨噬细胞的NF-κB核转位；Corilagin可明显下调巨噬细胞中TNF-α、NF-κB、MMP-1, -2, -9蛋白质和mRNA的表达。分子机制上，Corilagin通过靶向CD40-CD40L信号通路调CD40-CD40L/TRAF-2/NF-κB，明显下调斑块组织和巨噬细胞中CD40、CD40L、TRAF-2和NF-κB的蛋白及mRNA表达，Corilagin抗AS、抗炎及抑制巨噬细胞泡沫化的机制可能与调控CD40-CD40L/TRAF-2/NF-κB信号通路密切相关。研究结果进一步揭示了Corilagin干预治疗AS易损斑块的药理学基础及分子机制。