基于RANK与Wnt信号通路研究不同物理因子配伍治疗膝骨关节炎的作用与机制

We proposed several physical agents (low-frequency pulse ultrasound, microwave, low-level laser, low frequency pulsed electromagnetic fields, whole-body vibration therapy, extracorporeal shock wave therapy) as the effective modalities for the treatment of knee osteoarthritis focusing on the main pathological features based on our previous studies. The hypothesis of the project is: the combinations of different physical agents can effectively promote subchondral bone remodeling, cartilage protection and reducing synovitis. RANKL / RANK / OPG signaling pathway and the Wnt / β-catenin signaling pathway plays a key regulatory role in the treatment process. The project intends to adopt a comprehensive method including multi-level (overall, organs, cells, molecules) and multi-technological (high-resolution MRI, microscopic CT, flow cytometry, molecular biology, immunohistochemistry, etc.) ways to explore effects on physical structure and function, daily activities and participation of patients with knee osteoarthritis, as well as to investigate the mechanisms and effects of physical agents on articular cartilage, subchondral bone and synovitis of rabbits with knee osteoarthritis from the aspects of signaling pathways. Finally, we also intend to explore the mechanisms and effects on metabolism and apoptosis of osteoblast and osteoclast from the perspective of signal pathways.

基于不同物理因子（低频脉冲超声，微波，低功率激光，低频脉冲电磁场，全身振动治疗，体外冲击波治疗）针对膝骨关节炎的主要病理特征以及每种物理因子的有效性，同时结合课题组前期研究成果，本项目提出研究假设为：不同物理因子配伍能够有效地促进软骨下骨重塑、软骨保护以及减轻滑膜炎症。RANKL/RANK/OPG 信号通路和Wnt/β-catenin 信号通路在其治疗过程中发挥关键性调节作用。本项目拟采用多层次（整体、器官、细胞、分子）和多技术手段（高分辨 MRI、显微 CT、流式细胞仪、分子生物学、免疫组织化学等）相结合的方法，多角度探讨不同物理因子配伍对膝骨关节炎患者身体结构和功能、活动和参与，以及从信号通路的角度探讨不同物理因子配伍对兔膝骨关节炎的关节软骨、软骨下骨、滑膜炎症的影响与作用机制。最后，从信号通路的角度探讨不同物理因子配伍对成骨细胞和破骨细胞代谢与凋亡作用的分子学机制。

“慢性低度炎症”是膝骨关节炎重要的发病机制之一，其中TNF-α、IL-6及下游所影响的NF-κB、Wnt/β-Cantenin通路是治疗膝骨关节炎重要靶点。本研究通过联合不同参数的低频脉冲电磁场、全身振动疗法发现：1、75 Hz, 1.6/3.8 mT；8 Hz, 3.8 mT低频脉冲电磁场能够有效保护DMM所致小鼠软骨损伤，降低循环炎症因子，改善软骨下骨重建；2、振动频率5/10Hz，振幅4mm，重力加速度0.3g全身振动治疗对DMM所致的小鼠膝骨关节炎模型具有有限的保护作用；3、低频脉冲电磁场主要通过抑制细胞凋亡、TNF-α、IL-6分泌等途径，抑制NF-κB、Wnt通路，激活PPARγ通路，改善软骨及软骨下骨合成-分解代谢失偶联；4、全身振动疗法改善KOA软骨损伤可能与TNF-α、IL-6诱导的信号通路相关；5、脉冲电磁场与全身振动联合治疗对KOA软骨及软骨下骨重建保护具有一定叠加作用；6、8 Hz, 3.8 mT低频脉冲电磁场通过下调TNF-α、IL-6抑制GIOP小鼠骨细胞凋亡，改善骨代谢异常，抑制骨量流失，促进骨形成。通过此研究，本课题团队先创性从炎症信号通路角度探讨不同物理因子配伍对小鼠膝骨关节炎的关节软骨、软骨下骨及对骨代谢的影响与作用机制，为不同参数物理因子配伍联合治疗膝骨关节炎提供了有力生物医学证据，助力其向临床应用。