基于血清药物化学和网络药理学方法研究榅桲果实总黄酮抗动脉粥样硬化活性成分及作用靶点

Atherosclerosis is a kind of vascular degenerative diseases caused by comprehensive reasons such as lipid metabolism disorders, inflammation, oxidative stress, endothelial injury, which is also one of the major pathogenesis of cardiovascular diseases. The fruits, leaves, roots and seeds of Cydonia Oblonga Miller have long been used to prevent or treat cardiovascular diseases in traditional Uyghur Medicine. In our previous studies, we demonstrated that the extracts of Cydonia Oblonga Miller fruits have antihyperlipidemic, antithrombotic, anti-inflammatory, vascular endothelial protection and antioxidant effects, and flavonoids might be the main active components. Therefore, it is speculated that flavonoids of Cydonia Oblonga Miller has anti-atherosclerotic effect by different ways and the effective components could have several targets in different pathways, which might regulate and connect with each others like a network. In this project we plan to screen the anti-atherosclerotic effective components by serum pharmacochemistry and comprehensive assessment of different indexes. The tagets and signal pathways will be predicted by Network pharmacology to explore the mechanism. This study should provide experimental evidence for the clinical application of Cydonia Oblonga Miller, our results will provide novel experimental and mechanistic data for future development of Cydonia Oblonga Miller and establish a platform of Network pharmacology for research and development of other traditional Uyghur Medicine.

动脉粥样硬化是由于脂质代谢紊乱、炎症反应、氧化应激、内皮损伤等多种因素综合作用引起的血管退行性病变，是导致心血管疾病的重要病理基础。榅桲是传统维吾尔医学防治心血管疾病常用药物，其果实、叶、根、种子均可入药。本项目组前期研究发现新疆榅桲果实提取物具有调血脂、抗栓、抗炎、血管内皮保护和抗氧化作用，并且黄酮类化合物可能为主要有效组分，推测榅桲总黄酮可能通过多条途径发挥抗动脉粥样硬化的作用，并且可能包含若干活性成分的多靶点的整合调节作用。因此本项目拟采用血清药物化学和多药效指标综合评价相结合的方法，筛选榅桲果实总黄酮中抗动脉粥样硬化的活性成分，并利用网络药理学方法预测其可能的作用靶点及其作用网络，进一步探讨所调控的主要分子信号通路，阐明其作用机制。本项目研究结果可为榅桲临床用药提供科学依据，也为榅桲的开发利用奠定一定工作基础，更为维吾尔药材的研究开发建立网络药理学研究平台。

动脉粥样硬化（atherosclerosis，AS）是因脂质代谢紊乱、炎症反应等因素引起的主动脉慢性进行性病变，是多种心血管疾病的病理基础。研究证实榅桲提取物具有降低血压、调节血脂、抗氧化应激等功效，但榅桲抗AS作用机制和物质基础尚未明确。本研究采用UHPLC-Q-TOF-MS/MS技术分析鉴定榅桲提取物化学成分，通过血清药物化学的方法确定其原型入血成分；通过网络药理学方法构建榅桲入血成分-作用靶点-蛋白互作网络，信号通路富集分析，预测榅桲入血成分抗AS作用机制；建立AS动物模型确定榅桲抗AS药理作用，从调血脂、抗氧化、抗炎症反应、血管保护功能等多个方面深入阐明其作用机制。用代谢组学的角度研究榅桲总黄酮对动脉粥样硬化apoE-/-小鼠血清和尿液内源性代谢物的影响，并探讨其改善动脉粥样硬化作用的内在机制。血清药物化学研究结果表明，榅桲提取物鉴定出72种化学成分；原型入血移行成分有14种。网络药理学预测结果表明，抗AS主要靶点为IGF1R、EGFR、AKT1、PIK3CB、GSK3β、IL6、MAPK3、JAK2、PIK3CA等基因；抗AS主要信号通路为PI3K/AKT、Metabolic、VEGF、HIF-1、MAPK等信号通路。代谢组学及生物学验证结果表明，榅桲对AS动物模型具有一定抗AS作用，可降低动脉壁厚度，减少斑块；降低主动脉内膜厚度，减少平滑肌细胞增生、钙化、紊乱等病理过程；调节脂代谢紊乱、抗氧化应激、抗炎、保护血管内皮功能的作用，其作用可能与下调PI3K/Akt/GSK-3β信号通路，发挥抗炎、抗凋亡、调节能量代谢功能有关。