从VEGFs/VEGFRs信号转导及血管新生探讨温阳活血解毒组方对动脉粥样硬化易损斑块稳定作用

Cardiocerebreal vascular disases caused by Atherosclerosis have an important position in the reasons of people death. On the basis of our earlier researches, preliminary experiment has been carried out,proving that WenYangHuoXueJieDu prescription has the effect on anti-inflammation, on regulating the function of vascular endothelia, on suppressing intraplaque vascular angiogenesis and on stablizing vulnerable plaque. In this study, ApoE deficency mice model, Chicken chorioallantoic membrane model(CAM) and Human venous endothelial cell biocytoculture (HUVEC) were chosen and a series of experiments, both in vivo and in vitro, were designed. To evaluate the effect of WenYangHuoXueJieDu prescription on curing atherosclerosis, the degrees of inflammatory cytokines infiltration, intraplaque neovacularization, VEGFs/VEGFRs signaling pathway and stablity of atherosclerotic vlunerable plaque were observed. VEGFs/VEGFRs singaling pathway was emphasized to investigate its mechanism of stablizing the vulnerable plaques. Through this study, which bases on the innovative theory of traditional chinese medical pathology and therapeutic methed, we attampt to provide the theoretical and exprimental basis that WenYangHuoXueJieDu Therapeutic Method can cure atherosclerosis, to provide new orientation of preventing cardiovascular diseases by using chinese medicine.

动脉粥样硬化所致的心脑血管疾病在人类死亡原因中占有重要的地位。我们在前期研究基础上，初步预实验证实温阳活血解毒中药复方，具有抗动脉粥样硬化的炎症反应，调节血管内皮功能抑制血管新生，稳定粥样易损斑块的作用。本课题拟通过ApoE-/-小鼠动物模型、鸡胚绒毛尿囊膜模型联合体外人静脉血管内皮细胞培养，观察温阳活血解毒中药复方对动脉粥样硬化内斑块炎症浸润水平、斑块内血管新生程度、VEGFs/VEGFRs通路及斑块稳定性的影响，并着重从VEGFs/VEGFRs分子信号通路探讨其作用机制，为温阳活血解毒法治疗动脉粥样硬化提供理论和实验依据。这一立足于创新的中医病机理论与治疗方法的基础研究，将为中医药防治动脉粥样硬化及稳定易损斑块提供夯实的理论基础，为中医药防治冠心病提供新思路、新方法。

动脉粥样硬化是冠心病的病理基础。动脉粥样硬化易损斑块破裂所导致的急性冠脉综合征严重威胁着人类的生命健康。温阳活血解毒复方在临床治疗冠心病中已经取得了一定的疗效，为进一步阐明其作用机制，本研究主要围绕炎症反应、斑块的稳定性、血管新生及其相关的VEGFs/VEGFRs信号通路分别从体内AS模型小鼠和体外血管内皮细胞两部分探讨温阳活血解毒复方对AS易损斑块的稳定作用。体内AS模型小鼠研究结果显示，温阳活血解毒中药复方能抑制AS模型小鼠血清中炎症因子（MMPs、IL-1、IL-8、TNF-a、HIF-1、VEGF）表达水平，同时能降低斑块的易损指数及斑块内的新生血管密度，但其对相关的VEGFs/VEGFRs信号通路的相关分子的蛋白、基因的表达水平无影响；体外血管内皮细胞研究结果显示，温阳活血解毒复方含药血清对 HUVECs的增殖、迁移及细胞管形成的功能均产生了一定的抑制作用，且能影响VEGFR-2的表达，整体下调VEGFR-2及其下游信号链的表达，特别是phospho-ERKl/2，从而起到抑制血管内皮细胞的增殖、迁移及分化的作用。因此可以推断，温阳活血解毒复方是通过降低AS斑块中的炎症反应、减少斑块内的血管新生，调节VEGFs/VEGFRs信号通路相关信号分子表达等多方面的协同作用起到稳定易损斑块的作用。