NLRP3炎性复合体相关网络在血管稳态与重构中的作用与机制研究

NLRP3 inflammasome is found to be capable of sensing the change of homeostasis and act as a sensor of metabolic stresses for vascular homeostasis disorder and vascular remodeling. In previous work, we found that the genetic network constituted with the NLRP3 inflammasome and its down-stream genes may be the core of the system which maintenance the vascular homeostasis and remodeling. Risk factors activate the NLRP3 inflammasome related network in cells which form the main components of vascular microenvironment, then the activated NLRP3 inflammasome related network affects the cells in blood and the wall of vessel and play role in the vascular homeostasis and disorder and vascular remodeling. In present study, we will construct the inflammasome related network by using bioinformatics methods and analysis the difference expression of network members in cells of vascular microenvironment in vitro. To illuminate the biological mechanism of NLRP3 inflammasome related network in the development of vascular homeostasis disorder and vascular remodeling, we will observe the effect of the NLRP3 inflammasome on the vascular homeostasis in vitro/vivo and study the role of key factors in the network. Furthermore, we will screen the up- and down- stream genes of the NLRP3 inflammasome and complete the network, construct the point (genes)-line (gene-gene epistatic interaction) –network (gene interaction network)-system (vascular homeostasis and remodeling regulation system) model, identify the core elements that maintenance the vascular homeostasis and that induce the early vascular lesions, clarify the biological effects of NLRP3 inflammasome during the development of vascular homeostasis disorder and identify potential diagnostic markers and intervention targets.

NLRP3炎性复合体是感知内环境变化、介导血管系统自稳态失衡和血管重构的“传感器”。前期工作中，我们发现调控、编码NLRP3炎性复合体的基因与其下游基因共同构成的网络，可能是维持血管稳态与重构的关键节点。危险因子可激活构成血管稳态微环境主要细胞中的NLRP3炎性复合体相关网络，通过影响这些细胞的行为参与血管稳态维持、失衡及血管重构。本研究以NLRP3炎性复合体相关网络为切入点，分析其在构成血管稳态主要细胞中的差异；研究其在血管稳态从平衡到失衡过程中作用及关键分子，明确其介导血管稳态与重构的生物学过程；筛选NLRP3炎性复合体的上、下游相关基因，构建调控血管稳态与重构的点（基因）-线（基因-基因相互作用）-网（基因互作网络）-系统（血管稳态与重构调控系统）模型，并鉴定维持血管稳态、或在早期血管病变中发挥作用的核心元件。以期揭示血管稳态失衡与血管重构的生物学机制，发现疾病早期诊断和治疗靶标。

NLRP3炎性复合体是固有免疫应答的重要组成部分，也是感知机体代谢紊乱引起的内环境变化、介导血管系统自稳态失衡和血管重构的“传感器”。为研究NLRP3炎性复合体及其基因调控网络在相关疾病发病机制中的作用，我们应用RNAi技术在生理和ox-LDL刺激的病理条件下构建其效应蛋白低表达的细胞模型，通过全基因组表达谱芯片检测发现了63个受炎性复合体调控的差异基因，并在ApoE-/-小鼠中明确了与动脉粥样硬化相关的18个下游基因；发现2个随疾病进程加重而表达逐渐变化的基因，通过转录调控区分析与细胞学实验发现转录因子AP-2a和PPARG可参与调控上述基因表达，炎性复合体效应蛋白Caspase-1可以通过剪切AP-2a和PPARG，降低其与调控区的结合活性，进而调控下游目标基因的表达；确定了1个与ApoE表达密切相关的基因和1个与高脂饮食密切相关的基因，目前正在对上述基因导致动脉粥样硬化的机制与干预作用进行研究；通过基因组学研究发现一种可用于临床的降低血清LDL的分泌性蛋白。上述研究结果已达预期目标，在国际期刊发表4篇学术论文，申请1项国内专利，为深入研究NLRP3炎性复合体及其相关网络在维持血管稳态和参与动脉粥样硬化进程的分子机制提供了基础。