基于调控TLR4/MYD88/NF-κB信号转导通路探讨经筋微创松解疗法治疗膝骨关节炎的机制

In the research of meridian system, the Meridian-sinew is one of important contents.Clinical and animal experiments have been conducted on the treatment of Meridian-Sinew Release confirming that it can relieve sense of pain, swell and joint dysfunction of patients with knee osteoarthritis (KOA) , lower the levels of IL-1β, TNF-α, MMP-13, inhibit synovial inflammation, reduce articular cartilage destruction. However, we currently know little about the molecular mechanism of Meridian-Sinew release for the treatment of knee osteoarthritis.In the preliminary experiment, we found that the content of NF-κB had significantly decreased in joint fluid of three KOA patients after Meridian-Sinew release therapy. NF-κB plays a pivotal role in the downstream of TLR signaling pathway, which activates the transcription process of inflammation, causes the release of inflammatory factor, destructs the articular cartilage. Thus, we speculated that the Meridian-Sinew release probably works through regulating the TLR4 signaling pathway, inhibiting the downstream factor of MyD88 and NF-κB, blocking the secretion of IL-1β, TNF-α, MMP-13, reduce articular cartilage destruction.To explore the mechanism of Meridian-Sinew release of KOA, we intend to study its influence of TLR4/MYD88/NF-κB signal transduction pathway and chondrocyte proliferation in vitro, using rabbit osteoarthritis model, RT-PCR, Western Blot and other technology.

经筋系统是经络研究的重要内容之一。前期研究证实经筋微创松解疗法可缓解膝骨关节炎（KOA）疼痛、肿胀、功能障碍，降低兔外周血和关节液IL-1β、TNF-α、MMP-13，减轻滑膜炎症，保护关节软骨。预实验发现经筋微创松解治疗后，KOA患者关节液中NF-κB含量明显降低。NF-κB在TLR信号下游通路中起枢纽作用，能激活炎症转录程序，导致炎性因子释放，破坏关节软骨。由此推测经筋微创松解疗法可能通过调控TLR4信号通路，抑制下游MyD88、NF-κB表达，阻断IL-1β、TNF-α、MMP-13分泌，减少软骨破坏。本研究拟采用兔骨关节炎模型，运用RT-PCR、Western Blot等技术，研究经筋微创松解疗法对TLR4/MYD88/NF-κB信号通路及体外软骨细胞增殖的影响，探讨其治疗KOA的作用机制。

基于前述TLR信号转导通路体系和经筋松解治疗对膝骨关节炎关节液NF-κB、IL－1β和TNF－α影响的结果，本研究采用KOA兔动物模型，通过光镜观察软骨HE染色病理切片发现经筋微创松解疗法可改善KOA兔受累关节软骨细胞排列，有利于软骨细胞修复及增殖，减少血管翳生成；运用Real-Time PCR、Western Blot法检测TLR4、MyD88、NF-κB基因和蛋白的表达水平，发现经筋微创松解疗法可有效降低KOA兔软骨中TLR4、MyD88、NF-κB的基因和蛋白的表达，证明了经筋松解治疗作用机制是通过调控TLR4信号转导通路，抑制下游MyD88、NF-κB，阻断炎性细胞因子的分泌，减少软骨破坏，从理论上揭示经筋微创松解对KOA的治疗作用机制，促进经筋微创松解疗法治疗KOA的推广应用，弘扬祖国医学中的传统外治方法。