类风湿关节炎NKp44细胞协同MDSCs诱导OCs分化及中药“骨灵丸”对其干预机制研究

类风湿关节炎（RA）是一种常见自身免疫疾病，常导致关节骨破坏。申请者前期承担的国家和广东省自然基金研究表明：RANKL-RANK-OPG系统调控破骨细胞（OCs）活化是RA骨破坏的主要环节，中药骨灵丸对RA有良好护骨疗效，但相关免疫细胞（MDSCs和NKp44/NKp46+NK细胞）对该系统的调节机制及骨灵丸的护骨内涵尚有待深入研究。在以上工作基础上，本项目拟：①建立RA MDSCs和NKp44细胞共培体，分析两者作用下OCs分化规律；②建立剔除MDSCs和/或NKp46+NK细胞的RA动物模型CIA，研究两者通过OPG系统调控OCs促CIA骨破坏的机制；③运用骨灵丸干预CIA，进一步丰富其通过NKp44细胞协同MDSCs诱导OCs分化的护骨作用内涵。总体目标是阐明MDSCs/NKp44细胞调控OCs骨破坏机制及丰富骨灵丸护骨作用内涵，为揭示RA骨破坏免疫调控机制和新治疗药物研发奠定基础。

Rheumatoid arthritis (RA) is a common autoimmune disease and often leads to the bone destruction in the joint. The applicant previously presided studies subsidized by the Natural Science Foundation of China and Guangdong Province had concluded that the regulation of RANKL-RANK-OPG on osteoclast (OCs) was the main mechanism during the bone destruction in patients with RA. Traditional Chinese medicine guling pellet has shown their therapeutic effect of bone protect in RA. But the regulatory mechanisms of guling pellet to RANKL-RANK-OPG system and immune cells (including MDSCs and NKp44 cells or NKp46+NK cells) are yet to be studied. Therefore, based on the preliminary research foundation, we design the experiment below. First, separate the MDSCs and NKp44 cells from RA patients, and then establish the co-culture system of those cells in order to explicit the interaction mechanism in process of OCs differentiation. Second, establish the animal model of RA, collagen-induced arthritis (CIA) model, and then deplete these two cells in order to confirm the role of those cells in bone destruction in the CIA model. Third, intervene the CIA model with guling pellet and clarify its bone protect mechanism during the therapy of RA. The aim of this study is to clarify the mechanism of MDSCs and NKp44 cells in RA bone destruction and to confirm the bone protect mechanism of guling pellet. All of those studies may provide the foundation for the clarification of immune regulation mechanism in RA bone destruction and the development of new therapeutic drug.