基于抑制血管新生的中药豨莶草抗类风湿关节炎药效物质基础和作用机制研究

Rheumatoid arthritis (RA) is a systemic immune disease and very difficult to cure，which is characterized by chronic synovitis and cartilage lesions of the joints. RA patients will have been treated effectively and relieved their pain in essence only by inhibiting or blocking angiogenesis, of which is the key in the pathogenesis of RA. Chinese medicine theories hold that RA belongs to the category of " arthromyodynia" and "dredging the meridians and eliminating arthromyodynia" is the core idea of Traditional Chinese Medicine (TCM) in the clinical treatment of RA. Siegesbeckiae herba has the unique effects in dispelling wind-damp and dredging meridians, and previous research studies showed that the extract of Siegesbeckia glabrescens had a certain inhibitory effect on the secretion of the inflammatory factor IL-6, which is an important upstream factor of the JAK/STAT signaling pathway and may have a regulation on the expression of VEGF. Therefore, we give priority that the effective components in S. glabrescens can reduce the release of inflammatory factors and inhibit the expression of VEGF through the JAK/STAT signaling pathway to achieve the anti-angiogenesis of RA. In this study, MH7A cells and CIA rats were used as the test model of RA angiogenesis to explore the regulation of effective components in S. glabrescens on the expression of inflammatory factors and VEGF-related genes in cells, proteins and whole animals, and ultimately reveal the pharmacodynamic substances and molecular mechanism of inhibiting RA angiogenesis.

类风湿性关节炎（RA）是一种以关节慢性滑膜炎和软骨病变为主的系统免疫性疾病，难以治愈。血管新生是RA病症的关键，只有抑制或阻断RA的血管新生，才能从本质上实现对RA患者的治疗，解除其痛苦。中医理论认为，RA 属于“痹症”范畴，“通经祛痹”是中医治疗RA的核心思想。豨莶草具有祛风除湿、通经络的独特功效，但药效物质基础及作用机制尚不明确。课题组前期研究表明毛梗豨莶的提取物对IL-6的分泌具有抑制作用，而IL-6是JAK/STAT通路的重要上游因子，对VEGF的表达具有一定的调节作用。因此，我们认为毛梗豨莶中的有效成分可通过JAK/STAT通路，减少炎症因子的释放，抑制VEGF的表达，从而达到抗RA血管新生的作用。本课题拟以MH7A和CIA大鼠为RA血管新生模型，在细胞、蛋白和整体动物等多方面探讨有效成分对炎症因子以及VEGF相关基因的表达调控，揭示毛梗豨莶抑制RA血管新生的药效物质和分子机制。

该课题对毛梗豨莶的化学成分及抗RA活性进行较为深入的研究。采用HPLC-DAD/LC-MS化学筛选和活性筛选双重导向，快速发现结构新颖的化合物；通过活性评价和作用机制研究，发现活性先导化合物。从毛梗豨莶中共分离鉴定91个化合物，其中新化合物49个，包含三个结构新颖的二聚体。涵盖了对映-海松烷型二萜类、贝壳杉烷型二萜类、吉玛烷型倍半萜内酯类、愈创木烷型倍半萜等多种类型，丰富了课题组构建的萜类化合物库，为毛梗豨莶的深入开发利用奠定物质基础。对分离的91个单体化合物，采用LPS诱导RAW264.7细胞和CIA小鼠模型进行了抗RA活性及机制研究，并探讨了构效关系。其中二萜类化合物豨莶精醇抗RA作用机制研究发现，它通过抑制IL-6/JAK1,3/STAT3轴激活的炎症反应和JAK1,3/STAT3下调MMP2和MMP9的表达来减轻软骨降解来达到抗RA的作用；吉玛烷型倍半萜内酯类化合物Siegesbeckialide I能够与IKKα/β发生直接作用，导致IKKα/β失活，通过抑制IκBα的磷酸化和降解，在NF-κB通路中表现出显著的抑制作用，两类化合物均有作为抗RA先导化合物进行深入研究的价值。本项目为毛梗豨莶的抗RA药效物质阐明研究奠定基础，为抗RA靶向药物的设计提供依据，在加快抗RA创新药物研究开发方面具有重要的科学意义。研究结果已发表SCI论文7篇，申请专利2项，培养2名博士生，4名硕士生。