艾灸调控类风湿关节炎免疫异常的PD-1/PD-L1信号机制研究

Moxibustion is effective for rheumatoid arthritis (RA) without toxic side-effects，can efficiently regulates abnormal immune of RA and inhabits synovial inflammation. Over activation of T cells is the first important step in RA abnormal immune. PD-1 is the second signal co-stimulatory molecule which is required in immune pathology key step of RA when T cells are activated.It plays a role of the negative regulation when combined with PD-L1 in the starting step and inhibits the proliferation of T cell secretion. Based on previous research of mechanism of moxibustion for RA, we confirmed moxibustion could effectively control MAPK and other abnormal immune signal transduction pathways of the synovial cells in RA and inhibit proinflammatory cells secreting, and based on the fact that PD-1/PD-L1 pathway regulates the proliferation secreting of T cells negatively, we hypothesized that anti-inflammation and immune regulation of moxibustion for RA may work through regulating PD-1/PD-L1 signal pathway and regulating proliferation and secreting of T cell further. Guided by acupuncture theory, we plan to carry out an experiment using moxibustion on BL23(Shen Shu), ST36(Zu San Li) of animals with experimental RA around the key immunopathology and its target effect. We consider "PD-1/PD-L1 signaling pathway - inflammatory reaction" as the principle line of this research, choosing PD-1,PD-L1 as the entry point of this rechearch, and using siRNA、RT-PCR and other techniques to explore that moxibustion regulate PD-1/PD-L1 signaling immune response mechanism of RA, in order to further reveal the mechanism of Moxibustion in treating RA.

艾灸治疗类风湿关节炎（RA）具有调节免疫异常、抑制滑膜炎症、无毒副作用的优势。T细胞过度活化是RA免疫异常和炎症病理的重要始动环节，PD-1是其活化需要的第二信号共刺激分子，与PD-L1结合后在起始环节发挥负性调控作用从而抑制T细胞增殖分泌。基于前期研究发现艾灸可有效调控RA滑膜细胞MAPK等信号转导通路，抑制促炎细胞分泌炎性因子的基础和PD-1/PD-L1信号通路负性调控T细胞增殖分泌的事实，提出“艾灸可能通过调控PD-1/PD-L1信号通路，抑制T细胞增殖分泌，从而实现免疫调节和抗炎效应”的假说，以针灸理论为指导，艾灸肾俞、足三里治疗实验性RA，围绕免疫病理关键环节及目标效应，以“PD-1/PD-L1信号通路-炎症反应”为研究主线，选择PD-1、PD-L1为切入点，运用siRNA、RT-PCR等技术，探索艾灸调控RA免疫PD-1/PD-L1信号机制，更深入揭示艾灸治疗RA的作用原理。

本课题以中医理论为指导，艾灸肾俞、足三里治疗实验性RA，围绕免疫病理关键环节及目标效应，以“PD-1/PD-L1信号通路-炎症反应”为研究主线，选择PD-1、PD-L1为切入点，运用siRNA、RT-PCR、ELISA、免疫组化、流式细胞检测等方法技术，探索艾灸调控RA免疫PD-1/PD-L1信号机制，深化艾灸治疗RA的原理研究，为临床运用灸法治疗RA提供更丰富的科学依据。研究结论：①艾灸治疗RA具有确切的抗炎效应，且该效应与其调控PD-1表达有关。②艾灸可以调控RA大鼠TCR信号通路相关因子表达，该效应与其调控PD-1表达有关。参与T细胞活化早期的PTK和细胞分子中，与该调控机制关系密切的是p56Lck、CD69和CD25。③艾灸治疗实验性RA抗炎效应的作用机制可能与艾灸调控滑膜组织mPD-1、mPD-L1表达有关。④艾灸对实验性RA大鼠PD-1/PD-L1信号通路具有负性调控作用。