基于NF-κB、AP-1信号探讨苗药金乌健骨方调控类风湿关节炎滑膜成纤维细胞增殖和凋亡机制

The joint synovial tissue is the target organ of rheumatoid arthritis (RA). There was fully research evidence that the signal of nuclear factor kappa B（NF-κB）and activator protein 1（AP-1）are the main "switch" that syonvial cells start pro-inflammatory cytokine. Activities for the signal pathway regulated the expression of inflammative gene and promoted synovial cells proliferation，bone destruction. Controlling the non-normal activation of NF-κB and AP-1 signal pathway and down-regulating the expression of inflammative gene may become new treatment targets with RA. Based on the Traditional Chinese Medicine(TCM) difficulty and bottleneck of RA treatment, we thought "deficiency of kidney and stagnation of pathogen" is key factors in clinic practice of TCM according to its process of occurrence, development and evolution with RA. Under the guidance of this theory, we taked GuiZhou Miao Nationality herbs mainly to compose JinWu JianGu recipe. On the basis of previous clinical and experimental research, we will establish the animal and cell models to study NF-κB and AP-1 signal genes on synergism of proliferation and apoptosis of synovial cells with RA. On that basis, observed the effect of JinWu JianGu recipe on NF-κB signal genes, P65/P50, IκBα protein and AP-1 signal（c-Jun、c-Fos）with synovial cells. It may promote Chinese herbal compound targeting therapy lay the foundation for molecular research of fields with RA.

关节滑膜是类风湿关节炎（RA）的靶器官。有充分研究证据显示核周因子κB（NF-κB）和激活蛋白1（AP-1）信号是滑膜细胞启动致炎因子的重要"开关"，其信号途径的激活调节炎性基因的表达，促进RA滑膜成纤维细胞（RA-SF）增殖和骨质破坏。抑制NF-κB和AP-1信号途径的非正常激活，下调炎性基因的表达，可能成为RA治疗的新靶点。基于RA治疗的中医难点和瓶颈，根据其发生、发展和演变过程，课题组提出RA发病的中医"肾虚邪郁"理论，结合苗族医学"痹毒"理论，以贵州苗药为主组成金乌健骨方，在前期研究基础上，建立RA动物和细胞模型，研究NF-κB和AP-1信号基因相互补充协同调控RA-SF分化、增殖和凋亡的机制。在此基础上观察苗药金乌健骨方对滑膜细胞NF-κB信号基因、二聚体P65/P50、抑制蛋白α（IκBα）及对AP-1信号（c-Jun、c-Fos）的影响，为推动民族药靶向治疗RA研究奠定基础。

有充分研究证据显示核周因子κB（NF-κB）和激活蛋白1（AP-1）信号是类风湿关节炎（RA）滑膜细胞启动致炎因子的重要"开关"，其信号途径的激活调节炎性基因的表达，促进RA滑膜成纤维细胞（RA-SF）增殖和骨质破坏。抑制NF-κB和AP-1信号途径的非正常激活，下调炎性基因的表达，可能成为RA治疗的新靶点。本课题以苗医药理论为指导，提出类风湿关节炎（RA）发病的苗医“痹毒”理论，根据苗医学“毒以去为法”的治疗原则，应用苗药组成金乌健骨方，结合课题组前期研究基础，从动物实验和细胞试验两方面阐释了NF-κB、AP-1信号在类风湿关节炎发病机制中的作用及苗药金乌健骨方的调控机制，为研发苗药金乌健骨方提供了新的药理作用及机制。研究结果显示NF-κB、AP-1信号及其相关基因在CIA模型大鼠关节滑膜及人RA滑膜细胞呈高表达，苗药金乌健骨方能够有效抑制CIA模型大鼠关节滑模细胞NF-κB、AP-1信号的蛋白活性，下调致炎因子TNF-α、IL-1、IL-1β、IL-6、IL-17的分泌，减轻关节滑膜炎症。在细胞试验研究中，苗药金乌健骨方含药血清同样能够降低NF-κB、AP-1信号的蛋白活性及相关基因的表达，促进滑膜细胞的凋亡。. 以上研究结果表明以苗医“痹毒”理论为指导下组成的苗药金乌健骨方能够通过降低CIA模型大鼠及RA患者关节滑膜细胞NF-κB、AP-1信号蛋白活性及其相关基因的表达，抑制RA关节滑膜炎症，为苗医药金乌健骨方治疗RA阐释了药理机制，为开发苗药新药奠定了良好基础。项目研究发表学术论文20篇，其中核心期刊14篇，参加学术交流12人次，培养硕士研究生5人，贵州省省管专家1名，贵州省千层次人才3名。