傣药豆豉姜中新颖二苄基丁烷型木脂素基于MEK1和Cathepsin K双靶点的抗RA作用机制研究

Rheumatoid arthritis (RA) is currently one of the most serious diseases to human health, and so far there is no efficacious remedy on RA. Synovitis and bone absorption are the main factors leading to the destruction of cartilage and bone in patients with RA. Dai medicine Litsea cubeba root has been found to have the effect of anti-RA based on the previous research, and the new type of dibenzyl butane lignans represented by the compound LCA are discovered to have significant anti-inflammatory and tartrate resistant acid phosphatase (TRAP) activities for the first time. Molecular docking showed that MEK1 and Cathepsin K may be its potential targets, suggesting that the ingredients could exert the double effect of anti-inflammatory and inhibiting bone resorption by regulating MAPK pathway and osteoclast function, and the new dual target mechanism of Litsea cubeba root is different from the traditional anti-RA drugs. The project are planning to: 1. bioactivity guided isolate the characteristic lignans of Litsea cubeba root and mass-produce the target active compound; 2. to apply various cell models and animal models to verify its in vivo. and in vitro. activities, and evaluate its efficacy systematically; 3. preliminary elucidate the mechanism of target compounds playing the role of anti-RA through the intervention of MEK1 and Cathepsin K combined with the surface plasmon resonance, cell heat transfer technology, RNA interference technology as well as other scientific methods and technologies. The implementation of the project will lay the foundation for the development and utilization of the Dai medicine Litsea cubeba root with anti-RA effect.

类风湿性关节炎（RA）是当前严重危害人类健康的疾病之一，目前临床上尚无特效药。滑膜炎症和骨吸收是导致RA患者软骨及骨质破坏的主要因素。前期研究发现傣药豆豉姜具有抗RA作用，且首次发现以LCA为代表的新颖二苄基丁烷型木脂素类成分具有显著的抗炎和抗酒石酸酸性磷酸酶（TRAP）活性，分子对接显示MEK1和Cathepsin K为其潜在作用靶标，提示该类成分可能通过调控MAPK通路和破骨细胞功能来发挥抗炎和抑制骨吸收的双重作用，具有异于传统抗RA药物的新的双靶点作用机制。本项目计划：1.对豆豉姜中的特征性木脂素进行筛选和制备；2、应用多种细胞模型及整体动物模型对其进行体内外活性验证，系统评价其药效；3、结合表面等离子体共振、细胞热转移、RNA干扰等技术，初步阐明目标化合物通过干预MEK1和Cathepsin K发挥抗RA 作用的机制。本项目的实施完成将为抗RA民族药豆豉姜的开发利用奠定基础。

类风湿性关节炎（RA）是当前严重危害人类健康的疾病之一，目前临床上尚无特效药。滑膜炎症和骨吸收是导致RA患者软骨及骨质破坏的主要因素。前期研究发现傣药豆豉姜具有抗RA作用，且首次发现新颖的二苄基丁烷型木脂素类成分具有显著的抗炎和抗酒石酸酸性磷酸酶（TRAP）活性，且该类成分可能通过调控MAPK通路和破骨细胞功能来发挥抗炎和抑制骨吸收的双重作用，具有异于传统抗RA药物的新的双靶点作用机制。本项目已按计划完成：1.对傣药豆豉姜进行活性导向分离，确定以LCA为代表的二苄基丁烷型木脂素为其抗RA活性成分。根据LCA活性位点进行结构修饰，得到30余种LCA衍生物；并应用全合成技术成功合成LCA；2.利用RAW264.7细胞、破骨细胞及成纤维滑膜细胞进行体外抗RA评价，采用耳肿胀、气囊滑膜炎棉、球肉芽肿和佐剂性关节炎模型动物进行体内评价，分别在抗急性炎症、慢性炎症、抗滑膜增生及骨质破坏方面，系统的证实了LCA及其类似物的体内外抗RA药效；3.发现LCA及LCA1通过抑制TNF-α诱导的滑膜细胞中Raf、ERK和p90RSK的磷酸化以及降低p-MEK和p-MSK1的水平来阻碍 MEK - ERK信号途径的激活，抑制Akt和MAPK信号，下调NFATc1和MMP-9，防止RANKL诱导的破骨细胞分化，从分子水平揭示LCA及其类似物抗RA机理。