基于水杨酸蓄积-花生四烯酸代谢网络的丹参、三七逆转阿司匹林抵抗作用机制研究

Thrombotic events still occur in aspirin-treated patients with coronary artery disease. Danshen, Sanqi clinically reversed AR, but the mechanism is unknown. Based on the early confirmation of relevance between salicylic acid accumulation (SA) and AR, this study would investigate the mechanism of AR induced by SA and reversed by danshen and sanqi ingredients focus on arachidonic acid metabolic network (AMN). Three different AR model were established on rat for the following study. The progress of SA induced by pathophysiological factors will be explored from the perspective of drug enzymes and transporters. Intravascular eicosanoids and key proteins in AMN will be investigated under SA state，followed by analyzing the correlation between concentration of salicylic acid together with eicosanoids in plasma and platelet function in the use of artificial neural network model. These are used to establish a scientific evaluation system to reveal the process of SA disrupting AMN to induce AR. Based on the above system, influence on SA and AMN caused by Danshen and Sanqi component will be detected in vivo. By selecting representative composition，the reversal mechanism of AR by traditional Chinese medicine ingredients will be researched in vitro from COX-1 COX-2 pathway. This study is expected to clarify the relationship between SA-AMN and AR as well as the reversal mechanism of Danshen and Sanqi, forming a theory of AR occurrence and treatment system, in help to improve the clinical diagnosis of AR and integrated administration of western medicine and traditional Chinese medicine.

阿司匹林抵抗（AR）严重威胁冠心病患者生命安全。丹参、三七临床可逆转AR，但机制未明。本项目在前期证实水杨酸蓄积（SA）与AR相关性的基础上，拟围绕花生四烯酸代谢网络（AMN）研究SA引起AR以及丹参、三七成分逆转AR的作用机制：利用三种AR大鼠模型，从代谢酶和转运体的角度研究病理生理因素诱发SA的机制；考察SA状态下血管内花生酸类物质群及其代谢网络中关键蛋白的水平，采用人工神经网络模型分析血浆水杨酸和花生酸类物质浓度与血小板功能的相关性，揭示SA扰乱AMN从而引发AR的过程，建立科学评估体系；基于上述体系研究丹参、三七组分在体内改善SA和调控AMN的作用，并优选代表性成分，在体外从COX-1和COX-2两条途径研究中药成分逆转AR的机制。本研究有望阐明SA-AMN与AR的关系及丹参、三七的逆转作用，形成一套AR发生和治疗的理论体系，为改进临床AR的诊断、分型和中西药合用方案提供参考。

阿司匹林抵抗（AR）严重威胁冠心病患者生命安全。丹参、三七临床可逆转AR，但机制未明。本项目在前期证实水杨酸（SA）蓄积与AR相关性的基础上，围绕花生四烯酸代谢网络（AMN）研究SA引起AR以及丹参、三七成分逆转AR的作用机制。. 研究发现，酸中毒模型和LPS致炎模型的老年雌性大鼠长期给予AS后均造成AR，合用丹参、三七组分均可有效逆转AR的发生。对上述 AR 大鼠监测其体内SA代谢、排泄行为的变化以及相应肝药酶、肾脏转运体的表达水平，发现AR大鼠体内的SA和SUA发生了明显的蓄积，且SA肝脏代谢和肾脏排泄途径均出现了明显的阻滞，合用丹参、三七组分均可明显改善SA和SUA的蓄积，促进SA的代谢转化和肾脏排泄。. 考察AR状态下血管内花生酸类物质群及其代谢网络中关键蛋白的水平，采用人工神经网络模型分析血浆SA和花生酸类物质浓度与血小板功能的相关性，建立了科学评估体系，方法特异性高、重现性好，为患者早期筛查AR风险以及研究中药逆转AR的机制提供了有效的工具。基于上述体系揭示了丹参、三七组分对体内花生酸类物质群水平及AA代谢网络中关键蛋白的调控作用，为开发预防和治疗AR的新靶标拓宽了思路。. 基于血小板功能和信息学工具在体外筛选有效逆转AR的中药代表性成分，进一步从COX-1和COX-2两条途径研究AR发生以及中药逆转AR的机制，评价网络中各个环节的贡献度。研究发现丹参、三七中的丹参酮IIA、丹酚酸A、人参皂苷Rg1可能是在体外对逆转AR贡献度最高的中药成分，而COX-1酶的乙酰化反应、PPARγ、TXAS和NF-κB是AS发挥抗血小板作用以及中药逆转AR的关键节点。. 本研究初步揭示了水杨酸-花生四烯酸代谢网络与AR的关系以及丹参、三七的逆转作用机制，利用信息学工具进行数据挖掘和分析，形成了一套AR发生与逆转的理论体系和诊治方法，为改进临床AR的诊断、分型和中西药合用方案提供参考。