艾灸调节类风湿关节炎模型大鼠TH17/Treg 细胞失衡及Galectin-9 介导的机制研究

The mechanism of Moxibustion treating rheumatoid arthritis is related with regulating immune cells imbalance. Th1/Th2 and Th17/Treg cell are imbalanced in RA. Previous research showed that Th1/Th2 imbalance in RA can be regulated bu Moxibustion,but the effect of Moxibustion on Th17/Treg is still unknown. So we hypotheses that correcting Th17/Treg imbalance is one of the treating mechanism of RA by Moxibustion.To study the effects of moxibustion on Th17/Treg imbalance and the regulation mechanism, so that to prove the hypothesis, we will set up experimental RA like rats model, observe the histopathology variety of joints, Th17/Treg numbers and functional change in peripheral blood and synovium of joints, and also the mRNA and protein expression in Galectin-9 mediated Gal-9/tim-3 and CARMA1/NF-κB pathway.Meanwhile, by blocking Galectin-9, tim-3, and CARMA1 by antibody or siRNA interference to testify the hypothesis referred above. Our research will be helpful for illustrating the mechanism of moxibustion treatment on RA by reguation of immune cells, and improve the clinical treatment offered by Moxibustion.

既往研究证实：艾灸具有“温经蠲痹”作用，能有效改善类风湿关节炎（RA）患者临床症状和相关理化指标。Th1/Th2、Th17/Treg 细胞失衡是RA显著的免疫学改变，纠正其失衡能抑制关节炎症反应。艾灸治疗RA的机制与调节Th1/Th2 失衡有关，而是否与Th17/Treg 失衡有关尚不明了。我们推测：调节Th17/Treg细胞失衡可能是艾灸治疗RA的新机制。在此基础上，本研究将建立RA 大鼠模型并予艾灸治疗，观察关节组织病理变化、外周血及关节滑膜Th17/Treg 细胞数量、分化及Galectin-9 介导的Galectin-9/Tim-3、TCR/CARMA1/NF-kB 通路相关信号分子表达，同时通过特异性抗体阻断和RNA 干扰技术进一步明确其作用，探讨艾灸对Th17/Treg 细胞失衡的影响及调控机制。研究将从调节免疫细胞失衡角度为揭示艾灸“温经蠲痹”治疗RA的机制提供新的实验依据。

艾灸疗法“温经蠲痹”，能显著缓解类风湿关节炎临床症状、改善关节炎症，但其抑制关节炎症反应的机制尚未完全明确。Th17/Treg细胞失衡是RA中显著的免疫学改变，纠正其失衡能抑制关节炎症，有利于疾病的转归。研究显示艾灸能通过调节RA中Th1/Th2细胞失衡而发挥疗效，却对后者的作用尚不明了，而这很可能是艾灸治疗RA的新机制。其调控可能是通过Galectin-9 介导的Tim-3依赖的Galectin-9/ Tim-3信号途径和非Tim-3依赖的TCR/CARMA1/NF-κB信号途径。.本研究按时间完成了RA实验动物模型建立、艾灸治疗、关节形态分析和组织病理学观察；观察Th17/Treg细胞失衡的变化；抗体阻断Galectin-9；流式检测Th17/Treg细胞分化、T细胞分离；RORγt、Foxp3、CARMA1、NF-κB mRNA及蛋白表达。研究结果明确了艾灸“足三里、肾俞”穴能改善胶原诱导的关节炎模型（CIA）小鼠关节红肿，促进抑炎细胞因子分泌、抑制致炎因子分泌，逆转关节滑膜的炎性改变；其机制可能与促进Galectin-9分泌、上调Foxp3//RORγt/CARMA1/NF-κB基因与蛋白表达、调节Th17/Treg细胞失衡有关。Galectin-9可能是介导Th17/Treg细胞再平衡的物质之一。这将为艾灸治疗RA的临床应用提供指导。