浸润性T淋巴细胞表达IRF-7对骨性关节炎微环境的调控作用与补肾活血中药干预的研究

The incidence of osteoarthritis (OA) is closely related to the body microenvironment.Its local inflammatory response prompted synovial deterioration, bone injury aggravation and pain increase. T lymphocytes maintain the immune balance by regulatory role of antigen persenting cells. OA patients activated T lymphocytes which infiltrate into synovial fluid, and play the function of immune inflammation, so that arthritis occurs and its deteriorated degree is increased. The subject is based on the abnormal activation of infiltrating T lymphocytes in OA patients. With the role of IFN-alpha/beta induecd by self-immunine, infiltrating T lymphocytes were activated and expressed key interferon regulatory factor 7.Inflammatory factor signaling pathways phosphorylated IRF - 7, and activated the NF-κβwhich prompted expression of interleukin 6,so mediated by degradation of microenvironment in OA.We will research abnormal activation of infiltrating T lymphocytes and express the IRF - 7 and analyze its phosphorylation to activate NF-κβsignaling. So we confirm activation of IL - 6 expression to activate Jagged - Notch signal, resulting from regulating environmental change and degradation.We will research nourishing kidney and activating blood recipe which inhibit IRF - 7 expression and the role of its molecular mechanism,so inhibit chondrocyte worsening course. Therefore it confirms molecular mechanism of reversal role of the traditional Chinese medicine and theory of TCM prevention in OA.

骨性关节炎（OA）的发病与机体微环境密切相关，局部炎症反应可促使滑液增加、骨损伤加重和疼痛增强。T淋巴细胞受机体抗原递呈细胞调控，骨关节炎患者T淋巴细胞被激活侵入到关节中，发挥着免疫炎症功能，而高度浸润T淋巴细胞释放大量炎症因子，促进关节炎发生和恶化程度增加。本课题基于OA患者中浸润性T淋巴细胞异常激活，受自身免疫诱导产生的IFN-α/β作用，激活浸润性T淋巴细胞表达关键的干扰素调节因子7，炎症因子信号通路磷酸化IRF-7，活化NF-κβ促使白介素6表达，从而介导骨性关节炎微环境恶化的这一假说，研究OA内浸润性T淋巴细胞活化并表达IRF-7，分析其磷酸化激活NF-κβ信号，从而明确IL-6表达活化而激活Jagged-Notch信号导致微环境改变和恶化，研究补肾活血中药抑制IRF-7表达及对其分子机制的作用，抑制这一微环境恶化进程，有利于明确该中药逆转OA的分子机制，为中药防治OA提供基础。

骨性关节炎（OA）的发病与机体免疫微环境密切相关，局部炎症反应可促使滑液增加、骨损伤加重和疼痛增强。T淋巴细胞受机体抗原递呈细胞调控，OA患者T淋巴细胞被激活侵入到关节中，发挥着免疫炎症功能，而高度浸润T淋巴细胞释放大量炎症因子，促进关节炎发生和恶化程度增加。本项目研究发现，OA动物模型的关节软骨发生了不同程度的破坏，同时还伴有滑膜组织增生肥厚，炎性细胞浸润等病理变化；而且p38 MAPK和NF-κB信号通路的活性明显被激活，MMP-13、IL-1β、IL-6水平也高表达，表明说明过度激活的免疫炎性反应可能是导致关节软骨破坏的重要因素。在OA关节软骨中与免疫相关的TLR4，MyD88，IRF-7蛋白表达水平均上调，进一步通过IRF7病毒转染T淋巴细胞（IRF7-Tc）与人软骨细胞共培养第后发现软骨细胞的生长受到抑制，并出现细胞凋亡或死亡，表明T淋巴细胞中IRF-7过表达可抑制软骨细胞生长，促进了其凋亡或死亡，可能与其介导的免疫炎性反应有关。在IRF7-Tc与软骨细胞共培养后，结果发现软骨细胞的IRF7、STAT3 、MMP-13 mRNA和炎性因子IL-1β表达水平明显上升了，并通过动物体内验证了IRF7过表达可促使STAT3的激活，促进MMP-13表达、并能促使IL-1β分泌而导致软骨细胞凋亡现象发生。补肾活血中药能增加OA模型的体重和脾脏指数，使造模降低的胸腺指数恢复至正常水平，并可减轻OA关节软骨结构破坏的范围和严重程度，滑膜炎症水平，并具有促进软骨细胞增生的作用，表明补肾活血中药缓解OA炎症反应与其调节机体免疫器官的功能密切相关。在分子机制研究方面，补肾活血中药可通过下调IRF-7蛋白的表达，抑制TLR4，MyD88蛋白的异常表达，进一步抑制p38 MAPK和NF-κB信号通路的活性，使IL-6、MMP-13分泌水平下降，阻断OA免疫炎性微环境的发生，进而缓解炎症并延缓关节软骨破坏。因此阐明补肾活血中药抑制OA机体内免疫炎性微环境恶化的分子机制，为中药逆转OA提供新的实验依据，也为防治OA提供新的思路。