Wnt/β-catenin 信号通路与TLR4串话在动脉粥样硬化单核细胞中的作用及护心康的影响

Inflammation plays an important role in the occurrence of atherosclerosis,and monocyte is the most important inflammatory cells of them. Our research shows that TLR4 expression is increased in atherosclerotic plaques , and exacerbateds inflammatory response . Huxinkang can inhibit the expression of TLR4, which play a role in inhibiting vascular inflammation . TLR4 and Wnt appear in the atherosclerotic plaque, and the expression was all increased, suggesting the presence of crosstalk between them , but its internal relations and the mechanism is not clear. So, how the internal mechanism of crosstalk between the TLR4 signaling pathway and Wnt/β-catenin works, and how this mechanism in the inflammation of atherosclerosis acts. And what is the linkage between the role of Huxinkang in reducing the expression of TLR4 and this crosstalk. To clear these problems, this study intends to use transgenic animal model of atherosclerosis, by the technologysuch as specific antagonist,co-immunoprecipitation,real-time PCR, researching the role of crosstalk between Wnt/β-catenin and TLR4 in atherosclerosis monocytes,and the mechanism of Huxinkang in inhibiting the vessel wall inflammation.

炎症在动脉粥样硬化发生中起着重要的作用，单核细胞是其中最重要的炎症细胞。我们的研究表明，TLR4在动脉粥样硬化斑块中表达升高，加剧了炎症反应，护心康能抑制TLR4水平，从而起到抑制血管炎症的作用。TLR4和Wnt同时出现在动脉粥样硬化斑块中，且表达均升高，提示二者存在串话，但其内在联系和机制尚不明确。那么，TLR4和 Wnt/β-catenin信号通路之间串话的内在机制如何，该机制在动脉粥样硬化炎症中的作用如何，护心康降低TLR4的作用是否与二者的串话有关呢？为明确上述问题，本课题拟运用转基因动物复制动脉粥样硬化模型，通过特异性拮抗、免疫共沉淀、实时PCR等技术，研究TLR4和 Wnt/β-ca的串话在动脉粥样硬化单核细胞中的作用，以及护心康抑制血管壁炎症的机理。

本研究通过高脂饲料喂养复制TLR4基因敲除小鼠动脉粥样硬化模型及ox-LDL刺激单核细胞复制动脉粥样硬化细胞模型。采用病理组织学、特异性拮抗、免疫共沉淀、实时PCR等技术，研究了动脉粥样硬化发生时TLR4和Wnt/β-catenin的相互作用以及护心康对动脉粥样硬化的干预效应。通过研究发现，在动脉粥样硬化发生时TLR4和Wnt/β-catenin信号通路相关因子基因和蛋白表达升高，二者之间存在着相互作用和相互影响。护心康可以有效降低动脉粥样硬化发生时TLR4和Wnt/β-catenin信号通路相关因子的表达，从而起到治疗动脉粥样硬化的的作用。该研究丰富了中医活血祛痰及益气养阴治法的科学依据。通过研究，揭示了动脉粥样硬化的发病机制和调节靶点；阐明了护心康抗动脉粥样硬化的作用与Wnt/β-catenin和TLR4串话的关系，为动脉粥样硬化的防治和中医药研究提供新的思路和实验依据。项目研究过程中，发表学术论文9篇，参加国内学术会议5次，培养硕士生 7名，申报发明专利1项。