ALOX15介导的磷脂过氧化在“痰瘀互结”型冠心病的病理机制研究

" Binding of phlegm and stasis " is the main TCM syndrome of myocardial ischemic coronary heart disease, but its biomedical basis is still unclear.Our previous studies have found that when cardiomyocytes of animals on a high-fat diet showed significant lipid peroxidation under hypoxia, their cell membrane structure was damaged, and the mechanism might be related to the oxidase ALOX15.Therefore, we propose a scientific hypothesis that when cardiomyocytes are hypoxic, the fatty acid binding protein h-fabp recruits ALOX15 to the myocardial cell membrane and activates ALOX15, which induces phospholipid peroxidation and leads to increased sensitivity of cardiomyocytes to oxidative damage.During reperfusion, ROS enhances the interaction between ALOX15 and h-fabp, leading to an oxidative cascade of phospholipids in the myocardial cell membrane, thereby inducing cell death.This project intends to use the advanced technology of phospholipid oxidative metabolomics and recombinant protein to study the mechanism of phospholipid peroxidation by ALOX15 as a breakthrough, research on the biological basis of " Binding of phlegm and stasis " coronary heart disease. This may provide modern biological basis for early diagnosis, prevention and treatment strategies, drug research and new drug development for " Binding of phlegm and stasis " coronary heart disease.

“痰瘀互结”是心肌缺血型冠心病的主要中医证型，但其生物医学基础尚不清楚。我们前期研究发现：高脂饮食动物模型在心肌缺氧情况下，心肌细胞发生明显脂质过氧化，导致心肌细胞膜结构受损，发现其机制可能与氧化酶ALOX15介导的特异性磷脂氧化有关。因此，我们提出科学设想：在心肌细胞缺氧时，脂肪酸结合蛋白H-FABP募集ALOX15到心肌细胞膜并激活ALOX15，诱导磷脂过氧化，造成心肌细胞对氧化性损伤敏感性增加。在再灌注期，累积的ROS增强了ALOX15与H-FABP的相互作用，导致心肌细胞膜磷脂发生氧化级联反应，进而诱发细胞死亡。本项目拟采用磷脂氧化代谢组学和重组蛋白等先进技术，以ALOX15介导的磷脂过氧化作用机理为突破口，对“痰瘀互结”型冠心病的生物学基础进行研究，旨在为“痰瘀互结”型冠心病提供现代生物学证据，有利于其早期诊断、防治策略、药物研究及新药开发。

“痰瘀互结”是冠心病的主要中医证型之一，高血脂是主要危险因素，但其生物医学基础尚不清楚。在建立高脂饮食（High fat-diet，HFD）负荷左冠状动脉前降支（left anterior descending coronary artery，LAD）结扎术的“痰瘀互结”动物模型中发现，HFD增加了LAD结扎术的死亡率。由此，我们提出科学问题：在“痰瘀互结”型CHD中，HFD增加心肌缺血易感性的具体机制是什么？本课题采用多组学以及激光共聚焦成像等多种技术对科学问题进行了系统的研究。研究结果发现HFD可以增加心肌缺血的敏感性，而ALOX15介导的磷脂过氧化是其关键机制。这种损伤在缺血早期损伤就已经出现，并可能贯穿于冠心病缺氧/再灌注的整个疾病过程。此外，当HFD与ALOX15单独存在时并没有对心肌缺血表现出显著的易感性，而只有当HFD与ALOX15共同作用时才会引发心肌缺血的敏感性。由此，我们认为导致这种敏感性的原因可能是HFD为ALOX15提供了氧化底物，而ALXO15为氧化级联反应提供了催化酶，这也可能是“痰瘀互结”型CHD的关键物质基础。最后，采用丹蒌片和温胆片干预建立的“痰瘀互结”型CHD动物模型进行反向验证来更进一步阐明我们的研究设想。