Kv1.3通过MAPK/NF-κB途径调控巨噬细胞活化在血管再狭窄中的作用研究

Prevention and treatment of restenosis (RS) after percutaneous coronary intervention is an important project in the field of cardiovascular research. Macrophage migration and secretion of inflammatory cytokines play an important role in the occurrence of RS. Our previous study confirmed that the expression level of potassium channel Kv1.3 was significantly elevated in patients with acute coronary syndrome. We made a further study and found that Kv1.3 regulate the migration of macrophages probably through ERK pathway. However, the role of macrophage activation in the process of RS has not been reported. Here, we plan to make in vivo observation and in vitro mechanism study to clarify the role of Kv1.3 on the relegation of macrophage activation, confirm the regulating effect of Kv1.3 in MAPK/NF-κB pathways and explore the related mechanisms as well as determine the impact of Kv 1.3 on the macrophage activation and RS occurrence with the combined method of pharmacology and gene interference from molecular, cellular, and animal level. This project will contribute to the further understanding of the molecular mechanism of RS occurrence, and provide new strategies and intervention targets for the prevention and treatment of RS.

经皮冠脉介入术后再狭窄（RS）的防治是心血管研究领域的重要课题。巨噬细胞迁移和炎症因子分泌是导致RS发生的重要环节。本课题组前期研究证实钾通道Kv1.3表达水平在急性冠脉综合征患者中显著升高；进一步通过体外过表达和基因沉默Kv1.3通道，发现Kv1.3可能通过ERK信号通路调控巨噬细胞的迁移活动。然而Kv1.3调控巨噬细胞活化在RS过程中所发挥的作用及其机制国内外未见报道。本研究拟采用药理学与基因干预相结合、在体现象观察与离体机制研究相结合的研究模式，在整体动物、细胞和分子水平上综合验证Kv1.3对巨噬细胞活化的调节作用，探讨Kv1.3调控MAPK/NF-κB信号通路的机制，确定Kv1.3对MAPK/NF-κB的调节作用对巨噬细胞活化及RS发生的影响。课题的实施有助于进一步理解RS发生的分子机制，并有望为RS的防治提供新的策略和干预靶点。

钾通道Kv1.3是由6个α螺旋组成的4组蛋白质亚单位6次跨膜形成单孔道通道，不仅能够调节K+流动，而且通过蛋白-蛋白之间的相互作用参与细胞信号传递。既往研究提示Kv1.3参与巨噬细胞炎症活动，但是Kv1.3在血管再狭窄中的作用尚未明确。我们前期研究证实Kv1.3钾通道能够影响巨噬细胞的迁移活动，进而增加炎症细胞浸润，而这是促进血管再狭窄的重要原因。因此，我们提出如下假说：Kv1.3钾通道通过促进巨噬细胞活化，加重动脉内膜损伤，最终加重RS的发生发展。为了验证假说，我们从体内及体外水平进行深入研究。体内实验中，我们通过培育Kv1.3-/-ApoE-/-双敲基因小鼠，同时构建Kv1.3过表达慢病毒转染ApoE-/-小鼠，并对小鼠左侧颈总动脉完全结扎构建血管再狭窄模型，通过双向调节Kv1.3，观察其对血管再狭窄的影响。体外实验中，过表达和基因沉默Kv1.3探讨Kv1.3对巨噬细胞功能影响及作用机制。我们发现小鼠血管再狭窄模型中，Kv1.3加重血管再狭窄，而基因沉默Kv1.3后血管再狭窄降低；Kv1.3加重血管再狭窄的机制为通过调节MAPKs-NF-κΒ信号转导通路促进巨噬细胞趋化和炎症分泌活动。