ALDH3A1/4-HNE通路调控内皮细胞铁死亡在动脉粥样硬化发展中的作用机制研究

血管内皮细胞铁死亡过度激活可能加速动脉粥样硬化(AS)发生发展，但具体调控机制尚未明确。申请者前期研究发现：醛脱氢酶ALDH3A1基因敲除导致反应性羰基物质(RCS)如4-羟基壬烯醛(4-HNE)等沉积增加，谷胱甘肽过氧化物酶GPX4耗竭失活及铁死亡过度激活。据此推测，ALDH3A1通过定向代谢清除4-HNE，抑制血管内皮细胞铁死亡而发挥抗AS作用。本研究拟在已有工作基础上，利用CRISPR/Cas9和Cre/loxP等基因编辑技术，建立ALDH3A1、内皮特异性GPX4基因敲除斑马鱼与小鼠模型，同时运用生物信息学，表观遗传学及系统生物学等手段，进一步阐明血管内皮细胞铁死亡对AS发生发展的影响；探明ALDH3A1/4-HNE调控内皮细胞铁死亡的关键分子信号通路,及在AS发生发展中的潜在机制；观察调正细胞铁死亡对内皮损伤的保护和AS进展的抑制作用，为AS防治提供新的潜在干预靶点。

The overactivation of endothelial cell ferroptosis may play an important role in the formation and development of atherosclerosis (AS). However, the specific mechanism remains unclear. Our previous study showed that aldehyde dehydrogenase 3A1(ALDH3A1) knock out led to the accumulation of reactive carbonyl substances (RCS) such as 4-hydroxynonenal (4-HNE), depletion of glutathione peroxidase GPX4, and over-activated ferroptosis. Therefore, we speculate that ALDH3A1 may inhibit endothelial cell ferroptosis through 4-HNE detoxification, and play a protective role in AS. By using gene-editing technologies such as CRISPR/Cas9 and Cre/loxP, this study intends to generate the ALDH3A1 and endothelium-specific GPX4 gene knockout mutants in both zebrafish and mouse model. Through bioinformatics, epigenetics and system biology methods, we aim to further clarify the effect of vascular endothelial cell ferroptosis on the occurrence and development of AS; to explore the key molecular signalling pathway that ALDH3A1/4-HNE regulates endothelial cell ferroptosis, and the underlying mechanism in the occurrence and development of AS; to observe the protective effects of ferroptosis rescue on endothelial injury and AS progression, and provide new potential targets for the prevention and treatment of AS.