膜联蛋白A1调控炎性衰老在血管老化中的作用及机制

Inflammatory aging is one of the important mechanisms of vascular aging. The anti-inflammatory factor Annexin A1 (ANXA1) is associated with many vascular aging-related diseases such as atherosclerosis and diabetes. ANXA1 participates in the pathophysiological process of inflammation by binding to FPR2 and regulating NF-κB, which participates in the occurrence and development of inflammation-related diseases. However, the role of ANXA1 in the development of vascular aging is still unknown. Our previous study found that the expression of ANXA1 in plasma of healthy elderly, aged rat aorta or senescent endothelial cells was significantly lower than that of young adults, young rats and young adults, respectively.Thus, we propose the following scientific hypothesis: ANXA1 regulates the NF-κB signaling pathway by binding to FPR2 and inhibits the occurrence of inflammation, thereby delaying endothelial cell senescence and vascular aging. To verify this hypothesis, we conducted the following study.Fistly, we investigated the aging-related change in the plasma of ANXA1 and analysis the association between ANXA1 and age. Next, we knocked down the expression of ANXA1 in mice and HUVECs to investigate the process of endothelial cell senescence and vascular aging. Lastly, to reveal the mechanism, we treated ANXA1 knock down mice and HUVECs with hrANXA1 and the inhibitors of FPR2 and NF-κB. This study is of great significance for the provision of new theories and new targets for delaying vascular aging.

炎性衰老是血管老化的重要机制之一。抗炎因子膜联蛋白A1（ANXA1）与许多血管老化相关疾病如动脉粥样硬化、糖尿病有关，且可与FPR2结合调控NF-κB参与炎症的病理生理过程并参与炎症相关疾病的发生与发展，但是否与血管老化相关未知。我们在前期研究发现健康老年人血浆、老年大鼠主动脉和衰老内皮细胞ANXA1表达下降的基础上，结合上述认识，提出“ANXA1通过与FPR2结合调控NF-κB信号通路，抑制炎症发生，从而延缓内皮细胞衰老和血管老化”的科学假说。为验证上述假说，我们拟进行健康人血浆ANXA1增龄性变化研究，用ANXA1敲除小鼠和CRISPR/Cas9技术构建的ANXA1敲除人脐静脉内皮细胞及对敲除ANXA1基因的小鼠和内皮细胞分别予外源性ANXA1、FPR2抑制剂和NF-κB抑制剂等干预，以研究ANXA1在血管老化和内皮细胞衰老中的作用及其机制。本项目有望为延缓血管老化提供新理论和新靶点。

炎症与衰老关系研究是当前热点与重点，ANXA1是重要的抗炎物质，与许多心血管疾病相关，但与衰老关系尚未知。本项目探讨ANXA1对血管衰老作用及其机制，取得以下重要结果:1.健康老年人血浆ANXA1水平低于健康中青年人；2.老年小鼠主动脉ANXA1表达少于年轻小鼠；3.68周龄ANXA1敲除小鼠提前出现血管老化相关表型，如脉压增大、主动脉中膜和外膜面积增加、主动脉SA-β-Gal阳性表达及p21表达增多；4.ANXA1在HUVECs应激性衰老和复制性衰老过程中表达减少；5.敲低ANXA1的HUVECs表现出细胞衰老特征、功能受损，转录组学kEGG信号通路集中在细胞衰老通路，敲低ANXA1后通过补充回复ANXA1，可逆转细胞衰老现象，细胞功能出现改善,炎症因子表达下调，NF-κB转录因子表达下调。本项目研究成果已授权国家发明专利2项；发表论文4篇，其中中科院1区和2区文章各1篇；辅助培养硕士研究生1名（转博）。本项目成果为血管老化及其相关心脑血管疾病的防治提供新思路和新靶点。