基于Nrf2-ARE/NF-κB通路研究广东紫珠的抗炎药效物质及分子作用机制

Oxidative stress is a very important part of inflammatory response，so reducing oxidative stress level has become the important adjuvant means in the treatment of inflammatory disease. Although western medicine is remarkable to treat inflammation, Long-term use can cause serious adverse reactions. Therefore, natural anti-inflammatory drugs research is of great significance. Previous research has shown that phenylethanoid glycosides and flavonoids are the main ingredients of Callicarpa Kwangtungensis. Now five phenylethanoid glycosides have been isolated from the one anti-inflammatory effective parts of C. Kwangtungensis, and two compounds have shown good anti-oxidant and anti-inflammatory activity . Based on this, anti-oxidative stress is regarded as the key point in this project according to the structure characteristics of the main chemical compositions of C. Kwangtungensis, and the anti-inflammatory active ingredients of this species will be systematically isolated according to anti-inflammatory activity as indicator. Furthermore, the anti-inflammatory activity of the active ingredients will be investigated in vivo and in vitro via Nrf2-ARE anti-oxidative stress signaling pathway and NF-κB inflammatory signaling pathway. In addition, multiple target synergy anti-inflammatory effects of active ingredients are deeply discussed by the means of molecular docking and molecular dynamics simulation. The anti-inflammatory pharmacodynamic materials， structure-activity relationships, and molecular mechanisms of action will be illustrated guided by anti-oxidative stress. The accomplishment of this project will provide experimental and theoretical basis for the deeper use of C. Kwangtungensis in the treatment of inflammatory diseases in the future, which will further provide a beneficial exploration of medicinal plants about the genus Callicarpa in the treatment of inflammation from the point of antioxidation.

氧化应激是炎症反应重要组成部分，通过降低机体氧化应激水平来减弱炎症反应是当今医学领域的研究热点。长期使用西药治疗炎症可引发严重不良反应，因此天然抗炎药物研发具有重要意义。前期研究发现，苯乙醇苷和黄酮类成分是广东紫珠主要化学成分类型。目前从广东紫珠1个抗炎有效部位分离得到5个苯乙醇苷类成分，2个化合物显示了良好的抗氧化和抗炎活性。基于此，本项目根据广东紫珠主要化学成分的结构特征，选择抗氧化应激为切入点，以抗炎活性为指标，分离纯化广东紫珠抗炎活性成分，通过Nrf2-ARE和NF-κB信号通路探讨活性成分在体内外的抗炎活性，结合分子对接、分子动力学模拟等技术研究活性成分的多靶点抗炎作用，阐明以抗氧化应激为导向的广东紫珠抗炎药效物质、构效关系和分子作用机制。项目的完成将为广东紫珠在炎症疾病治疗方面的深层次开发利用提供科学依据，同时也为从抗氧化角度探讨紫珠属药用植物在炎症方面的治疗提供有益探索。

清热解毒中药广东紫珠在临床上对宫颈炎、上呼吸道感染、肺炎等具有显著疗效，氧化应激是炎症反应重要组成部分，通过降低机体氧化应激水平来减弱炎症反应是当今医学领域的研究热点。小鼠耳肿胀实验、小鼠腹膜毛细血管通透性实验、小鼠棉球所致肉芽肿3种炎症动物模型和血清NO抑制实验表明，广东紫珠醇提物对急、慢性炎症均有显著的抗炎疗效；从广东紫珠抗炎活性部位共分离鉴定得到42个化合物，主要结构类型分别为苯乙醇苷、黄酮和萜类。其中，黄酮和萜类是中低极性部位的主要化学成分；苯乙醇苷是中高极性部位的主要化学成分，在该药材中含量尤为丰富，可看做广东紫珠代表性成分；选择其中分离得到的6个苯乙醇苷对LPS诱导的RAW264.7小鼠巨噬细胞炎症模型进行研究，结果表明药物组对炎症因子TNF-α、IL-6、NO和活性氧ROS的产生均有明显抑制作用，呈一定量效关系；选择与炎症密切相关的氧化应激通路Keap1-Nrf2，进一步探讨2个苯乙醇苷类成分对Keap1、Nrf2及其下游抗氧化蛋白HO-1和NQO-1表达的影响，结果表明药物组可通过激活Keap1-Nrf2通路促进HO-1和NQO-1蛋白表达，从而抑制炎症因子释放；选择氧化应激为切入点，以人宫颈癌Hela细胞为模型，从细胞凋亡、周期阻滞及活性氧释放等方面初步研究了广东紫珠中苯乙醇苷和黄酮类成分在宫颈炎癌转化治疗过程中的治疗作用；采用光谱技术和理论模拟等方法分析了苯乙醇苷类成分在体外与人血清白蛋白的结合能力和微观结合模式，为探讨该类成分在生命体内的储存、运输、吸收、分布及代谢提供了实验依据。项目的完成为从抗氧化角度研究广东紫珠在炎症和宫颈癌治疗方面的作用提供了实验支持和理论依据。.已发表期刊论文5篇，会议论文1篇，另有1篇文章投稿到Journal of Ethnopharmacology（已接收），还有1篇论文正在整理中；申请专利1项；培养硕士研究生3名，其中1名已毕业。