促红细胞生成素通过GSDME介导的细胞焦亡抗动脉粥样硬化的作用机制

Atherosclerosis plays a vital role in the pathogenesis of cardiovascular disease, the exact mechanism of which remains underexplored. Pyroptosis, a special phenotype of programmed cell necrosis, was long regarded as caspase-mediated monocyte death in response to certain bacterial insults. Recent report has identified Caspase3/GSDME as a novel pathway leading to cell pyroptosis. It was speculated that pyrotosis might play a vital role in the progression of atherosclerosis, and aggravate plaque rupture, but the exact mechanism remains elusive. In the present study, we developed an EPO knockout zebrafish model with the help of CRISPR/Cas9 technology. Preliminary results showed that EPO knockout zebrafish protects the formation and progression of atherosclerosis by inhibiting pyroptosis via Caspase3 deactivation. Based on previous results，our project aims to further observe the function of pyroptosis in the formation and development of atherosclerosis protected by EPO. By analyzing pyroptosis makers in vascular endothelial cells, we try to explain the mechanism of Caspase3 deactivation and its possible relationship with GSDME cleavage. It is hoped to demonstrate the crucial role of pyroptosis in atherosclerosis protected by EPO and provide novel targets for the prevention and treatment of cardiovascular disease.

细胞焦亡作为炎性程序性细胞坏死，广泛参与动脉粥样硬化（AS）进展。Caspase3活化 GSDME是细胞焦亡的关键通路。既往证实，促红细胞生成素（EPO）在AS中发挥重要保护作用，但具体机制亟待阐明。本人前期研究发现，EPO基因敲除通过激活Caspase3，可能进一步剪切 GSDME引起细胞焦亡；胶原染色、免疫组化及电镜结果提示人AS斑块中存在细胞焦亡现象。本申请拟观察人动脉粥样硬化斑块，构建EPO基因敲除的AS斑马鱼模型，使用CRISPR、Cre-Flox等技术，评价动脉粥样硬化斑块中EPO及细胞焦亡标志物的表达及其与斑块不稳定的相关性；并通过调控Caspase活性及GSDME表达，观察EPO基因敲除引起细胞焦亡的信号通路及调节位点，阐明Caspase3/GSDME通路在EPO抗AS中的作用，探索外源性补充EPO阻断细胞焦亡对AS的保护，为AS的防治提供理论依据及治疗靶点。

细胞焦亡作为炎性程序性细胞坏死，广泛参与动脉粥样硬化进展。Caspase3活化GSDME是细胞焦亡的关键通路。既往证实，促红细胞生成素（EPO）在动脉粥样硬化中发挥重要保护作用，但具体机制亟待阐明。本研究在前期研究基础上，通过构建EPO基因敲除动物模型，进一步探索了细胞焦亡在 EPO 抗动脉粥样硬化发生发展中的作用，明确了Caspase3/GSDME 通路作为关键调控通路的主要机制，阐明了EPO介导糖尿病并发症进展的作用方式、作用区域以及代偿途径。同时通过结合临床群体资料分析，对EPO对动脉粥样硬化的保护作用提供了有价值的新线索，为后续的临床应用与特异性治疗奠定了理论基础。研究成果分别在Advance Science、Nutr Metab Cardiovasc Dis、 Cell Cycle等SCI期刊发表，获得发明专利2项，实用新型专利1项。相关研究成果多次在国际会议交流并获得美国心脏协会颁发的“国际青年研究者奖”。