防治动脉粥样硬化的中药：沙生蜡菊花降脂和抗炎活性成分的分析和鉴定

Atherosclerosis (AS) serious harm to human health. At present, the main treatment of AS is mainly Western medicine which have serious great side effects and can not cure the diease. Furthermore, these medicine may bring great damage on the body when long-term used. The essence of AS is the inflammatory lesions which participated by lipid composition. Therefore, the prevention and treatment of AS would be more effective if the medicine both have the effective of lipid-lowering and anti-inflammatory. Chinese medicine have the advantages of weak side effects and wide filter range. So far, there have no single herbs which both have the lipid-lowering and anti-inflammatory efficacy could be used for the prevention and treatment of AS. C-reactive protein (CRP) is the most strongest inflammatory markers in the inflammation process of AS. The pathway of CRP/CD32-NF-κB(p65)-VCAM-1 involved in the development of inflammation, and promote the formation of AS. We have found the lipid-lowering fraction of Flos Helichrysum arenarium. with anti-AS activity (HA) could reduce the NF-κB levels which induced by CRP for the first time, and this result suggesting that the HA may prevent AS by the dual role of lipid-lowering and anti-inflammatory. This project intends to take the CRP/CD32-NF-κB (p65)-VCAM-1 pathway as a target, and study the constituents in the HA, and investigated the regulation of HA and single compounds in the pathway. And then, clarify the material basis and mechanism of lipid-lowering and anti-inflammatory activity. This research would enhance pepole's understanding of the relationship between AS and the pathway above-mentioned, furthermore, provide new targets for the study and development of drug which could prevent AS.

动脉粥样硬化(AS)严重危害人类健康，目前其治疗以西药为主，缺点是副作用大，不能治愈，长期服用对机体损害大。AS本质是脂质参与的炎症性病变，因此降脂和抗炎并用防治效果会更佳。中药副作用较小，筛选范围广，值得探讨，但目前还没有兼具降脂和抗炎功效的单味中药防治AS。C反应蛋白(CRP)是AS炎症反应中最强的炎性标记物，且CRP/CD32-NF-κB(p65)-VCAM-1通路参与炎症的发生发展，可促进AS形成。我们首次发现沙生蜡菊花降脂部位（HA）具有抗AS活性，并能降低CRP诱导的核转录因子NF-κB水平，这提示HA可通过降脂和抗炎双重作用防治AS。本项目以上述通路为靶点，研究沙生蜡菊花降脂部位的化学成分，考察降脂、抗AS部位及单体化合物对该通路的调节作用，进而阐明沙生蜡菊花降脂和抗炎活性的物质基础和作用机制。此研究可增进人们对上述通路与AS形成之间关系的认识，为防治AS药物的研发提供新思路

脂质沉积和炎症反应是动脉粥样硬化形成的主要机制，本课题研究发现沙生蜡菊花醇提取物具有降脂和抗炎活性，推测其具有抗动脉粥样硬化作用。为阐明其抗动脉粥样硬化活性的物质基础及其作用机制，本项目对沙生蜡菊花降脂活性提取物的化学成分进行了系统研究，并从降脂和抗炎两方面研究单体化合物的抗动脉粥样硬化作用机制。项目采用常规提取、分离、结构鉴定技术，从沙生蜡菊花醇提取物中共分离、鉴定了72个化合物，其中21个新化合物，38个属内首次分离的化合物，进而明确了该活性部位的主要物质基础为黄酮和多酚类化合物。在此基础上，项目采用大鼠腹主动脉成功建立了可用于筛选抗动脉粥样硬化药物的血管环模型，该模型具有方便、快捷的优点，弥补了动物模型周期长、价格贵的缺点，分别根据血管形态学和其释放的炎症因子水平对模型进行评价，在动脉粥样硬化血管环模型中可见明显的血管损伤，并且出现血管内皮增生，此外通过酶联免疫反应检测损伤血管中炎症因子CRP、VEGF、NF-κB、JNK和P38均出现正相关、高表达现象，表明血管损伤即出现炎症反应。项目研究了沙生蜡菊花中的黄酮、黄酮醇、二氢黄酮类化合物对血管平滑肌细胞增殖、动脉粥样硬化血管环模型形态学、模型血管中炎症因子CRP、VEGF、NF-κB、JNK和P38表达以及化合物对LPS诱导的巨噬细胞释放NO的影响，来阐明沙生蜡菊花抗动脉粥样硬化机制。根据实验结果发现化合物Nariutin, Naringin, Eriodictyol, Luteolin, Galuteolin, Astragalin, Kaempferol的抗炎活性均强于阳性对照药心脑康，其中Eriodictyol，Astragalin，Kaempferol的抗炎活性强于其他化合物。同时该7个化合物均表现出较强的抑制LPS诱导的巨噬细胞释放NO的作用。其中Astragalin 不但能够有效抑制血管平滑肌的增生，还能降低血管环模型中CRP、VEGF、MAPK（JNK2）的表达，而从构效关系分析可推测黄酮类化合物的活性强于二氢黄酮类化合物的活性。此外，沙生蜡菊花中的主成分Astragalin表现出具有较好的降脂活性，由此表明Astragalin作为抗动脉粥样硬化先导化合物具有较好的开发价值。本课题的研究进一步证实了沙生蜡菊花具有防治动脉粥样硬化活性，其抗活性物质基础为黄酮类化合物，其作用机制为降脂和抗炎。