基于类风湿关节炎骨保护探究杜仲不同药用部位的性效机制

Based on our previous research, it is believed that Eucommia ulmoides can protect bones because it not only unblock meridians with its pungent property to inhibit bone damage caused by inflammation, but also promote bone repair with its sweet and warm properties. This research is designed to investigate the medicinal property difference between the bark, the leaf and the staminate flower of Eucommia ulmoides by observing their effects of inhibiting angiogenesis in synovial membrane, preventing bone damage, regulating bone metabolism and promoting bone repair. The mechanism of bone protection will be investigated through TNFα/NF-κB signaling pathway for the bark, the leaf and the staminate flower of Eucommia ulmoides by associating the traditional Chinese medicinal property theories and the contemporary pharmacological experiments. We will detect the ingredients of the three medicinal parts of Eucommia ulmoides in blood serum and perform pharmacodynamic research for these ingredients to investigate the essence of the sweet and pungent medicinal property of Eucommia ulmoides by associating its medicinal properties, effects and active ingredients. This research can provide theoretic basis for reasonable use of the three medicinal parts of Eucommia ulmoides. It is a good exploration for the substitution of the leaf and the staminate flower for the bark of Eucommia ulmoides, which is helpful for the protection of Chinese herbal resources and the expansion of Chinese herbal sources. This research is also an experimental basis for the future application of Eucommia ulmoides for treating the inflammatory bone damage.

在前期已有研究的基础上认为杜仲既可辛润通络，抑制炎症性骨破坏，又可甘温补骨，促进骨修复，从而起到骨保护作用。本项目拟通过动物实验，观察杜仲不同药用部位杜仲皮、杜仲叶、杜仲雄花提取物抑制类风湿关节炎模型大鼠滑膜血管新生，阻止骨破坏，以及调节骨代谢、促进骨修复的作用，以效测性，探讨杜仲3个不同药用部位的药性差异，将中药传统药性理论与分子信号通路研究相结合，从TNFα/NF-κB通路探究杜仲3个药用部位的性效机制。同时，应用血清药物化学方法检测杜仲皮、杜仲叶、杜仲雄花的入血成分，探求药效物质基础，从性、效、质三方面相结合探究杜仲辛甘的药性本质，为临床合理应用杜仲提供实验依据。杜仲叶与杜仲雄花的药性探索有助于完善杜仲药性理论，为杜仲叶、杜仲雄花代替杜仲皮入药提供有力支撑，并有利于挖掘药用资源，提高杜仲的药用价值和利用效率。同时，本项目也可为后期将杜仲应用于炎症性骨破坏的治疗研究提供实验基础。

研究结果显示，杜仲皮、叶、雄花的药效作用整体表现出相似的趋势。在体内可减轻CIA大鼠关节炎症，减轻骨破坏。体外抑制HFLS-RA的增殖迁移，抑制破骨细胞分化，促进成骨相关因子表达，抑制HUVECs的移行和血管新生因子的表达，抑制TNF-α/NF-κB通路相关蛋白。但杜仲皮、叶、雄花也表现出部分药效的差异。杜仲皮具有较好的骨保护作用，能更好抑制破骨细胞成熟，促进成骨相关蛋白表达；杜仲雄花在炎症抑制方面作用较强，且能促进滑膜炎细胞凋亡；杜仲皮与杜仲叶抑制NF-κB通路的激活、HUVECs迁移的作用强于杜仲雄花，但杜仲雄花能更有效地降低血管新生因子。.杜仲皮和雄花中8种化学成分整体含量高于杜仲叶。杜仲皮、叶、雄花中8种指标化学成分含量及分布差异显著，其中叶和雄花的差异更大。杜仲中桃叶珊瑚苷、京尼平苷酸、京尼平苷、京尼平、绿原酸和松脂醇二葡萄糖苷6种指标成分可以原型入血，并在不同药用部位之间表现出一定的药代动力学谢差异。.桃叶珊瑚苷在杜仲皮、叶、雄花中都有分布，具有抗炎、抑制破骨、促进成骨的作用，可能是杜仲干预RA的药效活性成分之一。.综合分析，杜仲皮应有辛味，有扶正和祛邪的双重作用。杜仲叶与雄花的药效与杜仲皮较为相近，可部分作为杜仲皮的替代品应用。杜仲雄花可能具有更相似的药性与功效，有潜在药用价值，可进一步进行研究开发和利用。.