复方中药单体涂层支架对微型猪冠状动脉内膜增生过程中TLR4-MyD88信号通路的影响

Aiming at the risks of 10％～20％restenosis and thrombosis in the drug eluting stent after which were implanted in coronary artery, we prepared the stents have been coated with Paclitaxel and Hirudin which come from Taxus Chinensis and leech respectively, the herbs that effects is for detoxicating, expelling wind-evil and removing extravasated blood, according to the theory of "Stirring of endogenous lateral wind"on atherosclerosis. Using the miniature swine coronary artery stent model, we prepare to study the inhibition of the stent to the injury , inflammation and endarterium hyperplssia of artery. At the same time, test the activation and expression of TLR4-MyD88 signaling relating genes. By using the methods of the MyD88 plasmid transfer in and MyD88 gene knockout (siRNA) in primary culture human coronary artery smooth muscle cell(HCASMC) respectively, then TLR4 agonist was used to activate the two group HCASMC, then the correlations were studied between the activation of TLR4-MyD88 signaling pathway relating downstream molecules and the proliferation and migration of coronary artery smooth muscle cells. The effect of Paclitaxel and Hirudin to the activated HCASMC and related indices were observed. It would be investigated that the effectiveness and molecular mechanism of the stents coated with Paclitaxel and Hirudin to prevent the restenosis and thrombosis in the drug eluting stent after PCI.

针对目前药物涂层支架仍有10％～20％ 的再狭窄率以及支架血栓形成的风险，我们依据动脉粥样硬化"络风内动"学说，选择具有解毒祛风、活血破瘀作用的红豆杉和水蛭，用其单体成份紫杉醇和水蛭素。在既往研究基础上, 制备成紫杉醇水蛭素涂层支架。采用微型猪冠状动脉支架置入模型，研究该支架置入对诱导血管损伤、炎症以及内膜增生过程的抑制作用，测定此过程中Toll样受体-4（TLR4)介导的MyD88信号通路相关信号基因的活化和表达。另用原代培养人冠状动脉平滑肌细胞（HCASMC)，分别在HCASMC内转染MyD88质粒和敲除MyD88基因，然后用TLR4激动剂诱导活化，明确TLR4-MyD88信号通路下游关联分子在不同条件下的活化状态与平滑肌细胞增殖、迁移等功能反应的相关性。观察紫杉醇水蛭素对上述活化模型及相关指标的影响，研究紫杉醇水蛭素涂层支架防治PCI术后再狭窄和支架血栓形成的有效性及分子机制。

针对目前药物涂层支架仍有10％～20％ 的再狭窄率以及支架血栓形成的风险，我们依据动脉粥样硬化“络风内动”学说，选择具有解毒祛风、活血破瘀作用的红豆杉和水蛭，用其单体成份紫杉醇和水蛭素。在既往研究基础上, 制备成紫杉醇水蛭素涂层支架。采用微型猪冠状动脉支架置入模型，研究该支架置入对诱导血管损伤、炎症以及内膜增生过程的抑制作用，测定此过程中Toll样受体-4（TLR4)介导的MyD88信号通路相关信号基因的活化和表达。另用原代培养人冠状动脉平滑肌细胞（HCASMC)，分别在HCASMC内转染MyD88质粒和敲除MyD88基因，然后用TLR4激动剂诱导活化，明确TLR4-MyD88信号通路下游关联分子在不同条件下的活化状态与平滑肌细胞增殖、迁移等功能反应的相关性。观察紫杉醇水蛭素对上述活化模型及相关指标的影响，研究紫杉醇水蛭素涂层支架防治PCI术后再狭窄和支架血栓形成的有效性及分子机制。研究结果表明：1.络风宁0号涂层复合物通过抑制血管局部炎症反应而起到防治PCI术后再狭窄的作用；2.络风宁0号涂层复合物通过抑制TLR4-MyD88-NF-κB信号通路而起到抗炎作用；3.络风宁0号涂层复合物通过作用于TLR4-MyD88-NF-κB通路中MyD88节点，促进MyD88蛋白在蛋白酶体途径降解，从而下调TLR4、NF-κB p65活性，减弱LPS启动的以IL-1β、IL-6、TNF-α为代表的炎症反应。