FcγRIIIA介导冠心病血瘀证的发病机制及川芎赤芍有效部位的干预效应研究

In previous study, we constructed the differential gene expression profile of coronary heart disease (CHD) with blood stasis syndrome by oligonucleotide gene microarray, which findings suggested the relevance of inflammation and immune responses to CHD with blood stasis syndrome at DNA level. These screened target genes were verificated by real-time reverse transcription polymerase chain reaction, including FcγRIIIA. Moreover, the regulatory role of FcγRIIIA was found in CHD with blood stasis syndrome by signal-net signal transduction network..In the present study, we will carry out the clinical correlation identification and source verification of target molecule FcγRIIIA. Basing on the results of validation, we will construct the cell system of human peripheral blood mononuclear cell-endothelial cell to explore the effect of the target molecule FcγRIIIA in monocyte-endothelial cell adhesion. Besides, we will make atherosclerosis model in apoE-knockout mouse to observe the effect of FcγRIIIA in atherosclerosis plaque stability and inflammatory cytokines expression. The chuanxiong and paeoniae radix effective parts was administrated in cell and mouse experiments to evaluate the effcts of traditonal medicine with promoting blood and removing stasis in anti-endothelial damage, lessening of inflammation and stablizing plaque, and to explore the role of target molecule FcγRIIIA in anti-atherosclerosis. Through these experiments, the present study will investigate the possible anti-atherosclerotic mechanism and effective targets of traditonal medicine with promoting blood and removing stasis.

课题组前期利用全基因组芯片技术，构建了冠心病血瘀证差异基因表达谱，从核酸水平揭示了冠心病血瘀证与炎症免疫反应的相关性，并对筛选出的目标基因（包括FcγRIIIA）进行了初步验证。并通过Signal-Net信号转导网络的构建，分析发现FcγRIIIA在冠心病血瘀证疾病过程中具有调控作用。.本研究在对目标分子FcγRIIIA进行临床相关性验证及来源鉴定的基础上，通过构建人外周血单核细胞-内皮细胞实验体系，探讨目标分子FcγRIIIA在单核-内皮细胞黏附中的作用，并通过构建apoE基因敲除小鼠动脉粥样硬化（AS）模型，观察FcγRIIIA表达对AS斑块稳定性以及炎症因子表达的影响；并通过观察中药的干预效应，评价活血化瘀中药川芎赤芍有效部位抗内皮损伤、减轻炎症反应、稳定斑块等抗动脉粥样硬化作用，及目标分子FcγRIIIA所扮演的"角色"，探讨活血化瘀中药抗动脉粥样硬化的作用机制及可能的作用靶点。

课题组前期利用全基因组芯片技术，构建了冠心病血瘀证差异基因表达谱，从核酸水平揭示了冠心病血瘀证与炎症免疫反应的相关性，并对筛选出的目标基因（包括FcγRIIIA）进行了初步验证。并通过Signal-Net信号转导网络的构建，分析发现FcγRIIIA在冠心病血瘀证疾病过程中具有调控作用。.本研究在对目标分子FcγRIIIA进行临床相关性验证及来源鉴定的基础上，通过构建人外周血单核细胞-内皮细胞实验体系，探讨目标分子FcγRIIIA在单核-内皮细胞黏附中的作用，并通过构建apoE基因敲除小鼠动脉粥样硬化（AS）模型，观察FcγRIIIA表达对AS斑块稳定性以及炎症因子表达的影响；并通过观察中药的干预效应，评价活血化瘀中药川芎赤芍有效部位抗动脉粥样硬化作用，及目标分子FcγRIIIA所扮演的角色。.研究结果发现，FcγRIIIA与冠心病血瘀证具有相关性，可能在冠心病血瘀证形成过程中发挥重要作用；冠心病血瘀证患者单核细胞处于活化状态，体内炎症水平特别是单核细胞来源的细胞因子处于较高水平；冠心病血瘀证患者高水平血清M-CSF能刺激单核细胞CD14+CD16+高表达，进而促进单核细胞CD14+CD16+表面CD54表达升高并产生大量TNF-α和IL-1前炎症因子，从细胞来源方面再次鉴定了目标分子FcγRIIIA与冠心病血瘀证密切相关；重组人M-CSF在体外能刺激人外周血单核细胞CD14+CD16+表达，IVIG能抑制该作用， 单核细胞CD14+CD16+能介导人外周血单核-脐静脉内皮细胞粘附；FcγRIIIA介导主动脉粥样硬化模型小鼠炎症相关因子释放增加，一定程度上影响小鼠主动脉粥样硬化斑块的稳定性；活血化瘀中药赤芍川芎有效部位能减轻炎症反应、稳定斑块、改善主动脉粥样硬化，其抗动脉动脉粥样硬化的作用靶点与FcγRIIIA有关。本研究基于冠心病血瘀证差异基因功能分析，探究活血化瘀中药抗动脉粥样硬化的作用机制及可能的作用靶点，为方证对应药物筛选和中药作用机制研究奠定基础。