化痰活血解毒方通过MAPKs信号通路抑制支架内新生动脉粥样硬化形成的机制研究

Percutaneous coronary intervention with stenting is the most widely performed procedure for the treatment of symptomatic coronary disease. Although drug-eluting stents (DES) have minimized the limitations of bare-metal stents (BMS), preventing the incidence of ISR in long-term after PCI is still a hot and difficult spot in the coronary intervention．Recent studies showed that in-stent neoatherosclerosis (ISNA) after DES was an important reason for the occurrence of late adverse cardiovascular events. The plaque rupture, intimal injury, eluting drug and stent struts target vessel after coronary artery DES implantation can induce and aggravate local inflammation reaction, which have important infection for activating of lipid laden macrophages gathered around stent struts, changing of smooth muscle cell proliferation, migration and phenotype, and promoting the formation of ISNA. The MAPK signal pathways are always running through the entire process of ISNA. "Sputum, silt, poison" is the etiology of atherosclerotic, pathogenesis of ISNA is the phlegm and blood stasis, heat coagulation. Huatan Huoxue Jiedu drugs can prevent the formation of ISNA by reducing inflammation through MAPK signaling pathway.Experiment is planed to perform in vivo and vitro to prove the hypothesis of Huatan Huoxue Jiedu drugs preventing ISNA.In vivo,we make rabbit model of atherosclerosis and DES implanting in abdominal artery and then to observe and study smooth muscle celland macrophagocyteby organ-cell-molecular level.In vitro,we culture smooth muscle celland macrophagocytewithHuatan Huoxue Jiedu drugsto explor the detailed mechanism of Huatan Huoxue Jiedu drugshow effecting the ISNA after stent implanting.

冠心病药物涂层支架（DES）置入后支架内再狭窄（ISR）可致严重心脏事件。研究显示支架内新生动脉粥样硬化（ISNA）是ISR的主要原因之一。DES置入后靶血管斑块破裂、内膜损伤、支架小梁及药物平台诱发血管持续的炎症反应，激活富含脂质的巨噬细胞在支架小梁周围聚集，平滑肌细胞增殖、迁移和表型改变，促使ISNA的形成；MAPKs信号通路贯穿其中。化痰活血解毒方具有抗炎、抗动脉粥样硬化的作用，但其对ISNA的作用及其机制不清。我们推测化痰活血解毒方可能通过MAPKs信号通路抑制ISNA。为此，本研究通过兔腹主动脉粥样硬化DES置入模型，以ISNA为靶点，从器官-组织-分子水平，观察化痰活血解毒方对ISNA的影响；利用巨噬细胞和平滑肌细胞，探索化痰活血解毒方通过MAPKs信号通路抑制ISNA的作用机制。为提高冠心病介入术后远期疗效提供理论依据和实验基础。

目前经皮冠脉支架置入术是治疗冠心病的主要手段之一，但研究发现DES置入后1年后的支架内的再狭窄率明显升高，病理学和影像学显示支架内新生动脉粥样硬化斑块（ISNA）是DES置入后再狭窄及支架内血栓的主要原因。研究认为支架置入处持续的炎症反应及涂层的抗组织增生药物损伤血管的愈合是导致ISNA的重要原因之一。MAPKs参与血管的炎症反应、血管的损伤后的修复，可能是ISNA形成的信号通路之一。中医认为ISR的主要病机是痰浊血瘀、热毒凝聚。化痰活血解毒治疗是预防支架内再狭窄的重要治则和治法。本研究通过动物实验过建立兔腹主动脉粥样硬化狭窄DES置入后再狭窄模型，观察化痰活血解读方干预24周后的病理形态学、影像学、血清炎症水平、凝血功能和信号通路相关蛋白的表达，明确化痰活血解毒方抗支架内新生动脉粥样硬化斑块作用。通过平滑肌细胞和巨噬细胞的实验，探索化痰活血解毒方通过MAPKs通路抑制平滑肌细胞和巨噬细胞功能而发挥其抗支架内再狭窄的作用机制。研究结果发现DES置入后24周，支架内管腔狭窄、新生内膜的显著增厚、发生新生的动脉粥样硬化斑块。化痰活血解读方能减少DES置入后血管的再狭窄率，减少新生内膜的厚度和面积，降低血清的炎症因子水平。化痰活血解毒方能显著抑制平滑肌细胞表型的转换，能减少巨噬细胞的募集，调节MAPKs相关通路蛋白的表达。此外，化痰活血解毒方能抑制LPS可诱导RASMCs细胞增殖和迁移，减少其凋亡，且能下调MAPKs信号通路中p-Erk1/2、p-p38 MAPK及p-JNK蛋白的表达水平。化痰活血解毒方也能抑制巨噬细胞的泡沫化水平，降低炎症反应水平。故推测化痰活血解毒方能够通过MAPKs信号通路调节平滑肌细胞和抑制巨噬细胞功能，发挥预防DES置入后支架内狭窄的作用机制。本研究为预防支架后再狭窄提供更多的治疗措施手段，为中医药在冠心病的治疗中的作用提供更多的理论证据和支持。