基于Notch/Treg信号通路研究中药新风胶囊改善佐剂关节炎大鼠肺功能的机制

Rheumatoid arthritis (RA) can not only lead to joint inflammation but also induce other damage. The decline of lung function is the primary lesions outside the joint damage in RA. For the lack of effective treatment means, it is necessary to reduce RA lung function system in-depth study. Chronic inflammation stimulation induced Notch express imbalances, which leads to the regulatory T cells (Treg) regulating immune dysfunction. The change of RA cytokine network cause immune complex deposition in the lung tissue to stimulate, eventually reduced lung function. It may be one of the RA reduced lung function mechanism. From regulating the Notch pathway for Treg regulate balance, thus improve RA lung function. Our research group according to the " treat arthralgia from the spleen " theory, propose the hypothesis that the Xinfeng capsule can intervene in the Notch pathway, regulate the expression of Treg and cytokine network, thus improving lung function of adjuvant arthritis (AA) rat. Through the in vivo and in vitro experiments, this study research that Xinfeng capsule can regulate the expression of Notch pathway and Treg, which reduce the immune complex deposition and inflammatory reaction, eventually improve lung function. It provides a experimental basis for the Chinese medicine improvement of RA lung function.

类风湿关节炎（RA）除关节炎症外，还可发生关节外病变，肺功能降低是RA关节外损伤首要病变，对其治疗缺乏有效的手段，有必要对RA肺功能降低进行系统深入研究。RA肺功能降低机制之一可能是慢性炎症刺激诱导Notch通路表达失衡，导致调节性T细胞（Treg）调节免疫功能紊乱，RA细胞因子网络改变，致使免疫复合物沉积对肺组织刺激，最终出现肺功能降低。基于以上机制，从调控Notch通路入手，寻求Treg调节平衡而改善RA肺功能。本课题组根据“从脾治痹”理论，提出采用“益气健脾通络”中药复方新风胶囊干预Notch通路，调控Treg表达和细胞因子网络，从而改善佐剂关节炎(AA)大鼠肺功能的假说。本研究通过在体、体外实验，研究新风胶囊通过对Notch通路和Treg的调节，降低免疫复合物沉积和炎症反应，最终改善肺功能。这将为中医药改善RA肺功能提供实验依据。

类风湿关节炎(RA)除关节病变外，还可引起其他脏器损伤。肺脏含有丰富的结缔组织和血管而成为RA关节外病变的首要靶器官。RA肺病变具有起病隐匿、病情进展快、病死率高的特点，成为RA首要死因。RA肺功能降低早于临床表现及胸部影像学异常。对RA肺功能降低进行研究尤为重要。Notch通路传导失衡导致调控Treg不足，促使免疫调节功能紊乱，导致RA肺功能降低。中医药在治疗RA肺病变中发挥重要作用。采用“益气健脾通络法”而拟定中药新风胶囊（XFC）治疗RA,疗效安全确切。新风胶囊能明显改善RA患者关节肿胀等病变，提高肺功能参数，改善RA干咳、咳痰、胸闷、气短等肺部症状体征。因此，从Notch信号通路、Treg角度研究XFC改善RA肺功能具有重要意义。. 本研究通过观察XFC对AA大鼠关节、肺功能变化、肺组织形态学的影响， XFC对AA大鼠血清细胞因子网络谱的影响，XFC对AA大鼠外周血Treg、肺组织CD4/CD25/FoxP3的影响，XFC对AA大鼠外周血CD4+Notch1+、CD4+Notch1+、CD4+Notch4+T细胞的影响，XFC对AA大鼠滑膜、肺组织Notch通路的影响。并研究XFC含药血清对肺间质成纤维细胞Notch通路、Treg调控作用机制研究。结果发现，XFC能升高IL-10、IL-35表达，降低IL-6、IL-17。XFC能降低CD4+CD25+ Notch3+Treg表达。免疫组化显示，XFC能降低CD4表达，升高FoxP3表达；Realtime-PCR法检测Notch通路相关基因发现。XFC降低Notch1、delta1和HES1的mRNA表达。Western Blot法检测Notch通路相关蛋白发现。XFC组降低Jagged 1、Jagged 2、Notch 4蛋白表达。XFC组能降低Ⅱ型肺泡上皮细胞Notch1、Notch3、Jagged1、HES1。说明XFC能升高外周血Treg细胞水平，抑制Notch/Treg信号通路过度激活，抑制免疫炎症反应，改善肺功能。. 本研究采用XFC治疗RA肺功能降低并探讨其作用机制，从而为开辟一条适合中医药治疗RA肺功能降低提供新途径。