NLRP3炎症小体在孕期炎症刺激导致子代动脉粥样硬化中的作用及其机制的研究

Inflammation is throughout the atherogenesis, and the incidence of Atherosclerosis (AS) traced back to inflammation during pregnancy is expected to expand the time window for prevention and treatment of AS. Our previous studies showed that typical pathological alterations represented the early stage of AS, and aortic dysfunction characterized by significantly higher vasoconstriction were both observed in the offspring rats from prenatal inflammatory exposure. Furthermore, the expression of inflammatory factors in aorta was obviously increased, and the NLRP3 inflammasome was exceptionally activated. Accordingly, we speculated that NLRP3 inflammasome activation should be the crucial molecular basis ,which triggers and maintains the non infectious inflammation in aorta and promotes AS. This project aimed to reveal NLRP3 inflammasome variations in offsprings responded to prenatal inflammation on the basis of LDLr-/- mice model induced by prenatal inflammation exposure, especially, effects of the key molecular of NLRP3 inflammasome on AS in offsprings were explored based on the double gene knockout mice models. Additionally, inhibitors, gene transfection, gene knockout technologies, etc, were employed to regulate the mitochondrial activity in vivo and in vitro, and thus the mechanism of NLRP3 activation was further elucidated from the pathway of mtDNA damage. This project would provide experimental and theoretical basis for new molecular targets, and develop new pharmaseutical approach in the treatment of AS.

炎症贯穿于动脉粥样硬化(AS)的始终。将AS发病追溯至孕期炎症刺激有望扩大AS预防和治疗的时间窗。前期研究表明孕期炎症刺激子代大鼠表现出早期AS的典型病理变化，及以血管收缩反应性显著增高为特征的血管功能失调，同时血管组织炎症因子表达显著增高，NLRP3炎症小体异常激活，推测活化的NLRP3炎症小体可能是孕期炎症刺激子代激发和维持非感染性血管炎症、促发AS的重要分子基础。本课题拟建立LDLr-/-小鼠孕期炎症刺激子代AS模型，以NLRP3炎症小体为切入点，应用双基因敲除动物模型等技术揭示孕期炎症刺激子代血管NLRP3炎症小体的变化规律，深入探讨NLRP3炎症小体各关键分子对子代AS形成的影响；采用抑制剂、基因转染、基因敲除等技术调控线粒体的体内、体外活性，从mtDNA损伤的角度揭示NLRP3激活的分子机制。本研究旨在为寻找新的AS有效防治靶点或防治药物提供实验和理论依据。

动脉粥样硬化(atherosclerosis, AS)导致的心血管疾病已成为全球最常见的死因,.传统观念中的危险因素并不足以完全解释AS发生。炎症过程贯穿于AS的始终，将AS发病追溯至孕期炎症刺激有望扩大AS预防和治疗的时间窗。本项目利用孕期脂多糖诱导孕鼠炎症模型，观察子代大鼠AS的发生与血管组织NLRP3炎症小体异常激活之间的关系。研究发现孕期LPS暴露子代大/小鼠血管组织损伤伴随炎症因子高表达，NLRP3炎症小体异常激活，同时线粒体功能障碍；子代叠加DOCA或高脂饮食，血管损伤、局部炎症、线粒体功能障碍及其NLRP3炎症小体活化会进一步加剧。孕期炎症暴露子代高脂喂养出现血脂代谢紊乱，体重异常增加。LPS刺激子代大鼠肝脏脂质代谢基因网络紊乱，高脂喂养使Vldlr和Tm7sf2基因的表达改变更为显著。体外实验通过mtDNA过表达和其抑制剂，证实mtDNA损伤是线粒体功能障碍、导致NLRP3炎症小体异常激活的原因。Apoe-/-caspase-1-/-小鼠孕期炎症刺激子代比apoe-/-小鼠孕期炎症刺激子代动脉粥样硬化病理改变明显减轻。本项目为本项目为基于NLRP3炎症小体等作为分子靶标的新药设计新策略奠定基础。同时，也可为其他成年慢性疾病的研究和防治提供有益借鉴。在该课题资助下，进行了较广泛的学术交流；培养硕士研究生5名；发表SCI论文4篇，中文核心期刊论文4篇；全国学术大会分会报告2次。