lncRNA-MEG3/miR-181/INPP5E调控网络在运动训练促进骨性关节炎康复中的作用及机制

Osteoarthritis (OA) is the most common joint disease that causes pain and dysfunction. Exercise training is effective in OA rehabilitation but the mechanism is unknown. It has recently been found that exercise training can promote autophagy of cells, and the occurrence of OA is closely related to autophagy. LncRNA is involved in the regulation of autophagy and abnormal expression in OA. Whether exercise training can affect cell autophagy by lncRNA is still unknown. The expression of lncRNA-MEG3 was inhibited and the autophagy was induced by low intensity stress stimulation in inflammatory chondrocytes. The chip and bioinformatics analysis showed that miR-181 related to lncRNA-MEG3 was up-regulated after stress stimulation and may affect autophagy related gene INPP5E expression. In this study, we will use body weight-supported exercise training and investigate the mechanism of LncRNA-MEG3/miR-181/INPP5E regulatory network by using gene expression regulation, luciferase reporter gene assay, RNA pull-down and RIP. Detect whether exercise training could affect the rehabilitation of OA and the potential mechanism through this regulatory network. This project will provide a new experimental and theoretical basis for further understanding the pathogenesis of OA and optimizing its treatment strategy.

骨性关节炎（OA）是引起疼痛和功能障碍的最常见关节疾病，运动训练对OA康复有效但机制未明。近来发现，运动训练可以促进细胞自噬，而OA的发生与自噬密切相关，lncRNA参与自噬的调控并在OA中异常表达，运动训练是否可以通过lncRNA影响细胞自噬尚未清楚。课题组前期发现，模拟运动训练力学信号的低强度应力刺激能抑制lncRNA-MEG3的表达，并促进炎症软骨细胞的自噬；芯片及生物信息分析发现与lncRNA-MEG3功能相关的miR-181在应力刺激后表达上调，并可能影响自噬相关INPP5E表达。本研究拟以减重运动训练为干预手段，运用基因表达调控、荧光素酶报告基因、RNA pull-down、RIP等技术探究LncRNA-MEG3/miR-181/INPP5E的网络调控机制，并探讨运动训练是否通过该调控网络影响OA的康复及可能机制，为深入了解OA的发病机制及优化其治疗策略提供新的实验和理论依据。

骨性关节炎是导致肢体肿痛及身体残障的退行性关节疾病，不仅严重影响患者的生活质量，也给社会造成巨大的经济负担，已成为一个全球性的公共卫生健康问题。目前针对OA的治疗方法以控制疼痛及病程终末期关节置换为主，尚缺乏治愈或阻断其病理进展的有效手段。WHO提出运动训练可作为防治OA的重要方式之一，近来发现，运动训练可以促进细胞自噬，而OA的发生也与自噬密切相关，lncRNA参与自噬的调控并在OA中异常表达，本课题通过建立软骨细胞炎症模型和创伤性骨关节炎模型，分别给予机械应力刺激和跑台训练，应用Western Blot、RT-PCR、基因沉默、荧光染色、流失细胞学等技术，探讨了机械应力刺激对软骨细胞增殖、凋亡、自噬、LncRNA MEG3等的影响以及减重跑台训练对创伤性骨关节炎大鼠关节软骨和软骨下骨的影响，结果表明：①机械应力刺激通过影响 LncRNA MEG3 的表达， 调控炎症环境下软骨细胞的自噬水平从而延缓软骨细胞的退变。②机械应力刺激以强度依赖的方式（1000μstrain、 2500μstrain、 4000μstrain）影响软骨细胞的增殖、表型和活力以及调控初级纤毛的发生率、长度以及自噬的表达， 此外 INPP5E 参与机械应力介导的初级纤毛和自噬的交互调控。③减重跑台训练减缓了创伤性骨关节炎大鼠关节软骨的退变和软骨下骨的骨质流失。初步阐明适度应力运动训练可以通过lncRNA-MEG3 相关的调控机制影响软骨细胞自噬，从而维持软骨细胞内稳态，防止细胞外基质进行性破坏和软骨退变，最终改善OA。该课题的研究结果对深入了解OA病理进展及康复的分子机理具有重要意义，为寻找OA治疗的有效靶点及进一步优化OA治疗策略提供了理论基础。