穿龙薯蓣皂苷对CIA鼠CD4+T细胞亚群平衡的影响及其调控机制研究

CD4+T cells play a critical role in the Rheumatoid Arthritis(RA), but the mutual dynamic of its subsets' function in RA and the relationship between the changes of the subsets and the RA's development is still not clear. This study is based on the latest research results about CD4+T cell subsets and the datas of Project from National Natural Science Foundation of China completed by our team last year. We plan to establish the CIA mouse models and observe the variation of Thl/Th2, Th17/Treg, their specific transcription factors, the proteins in their signal transduction pathway and their related cytokines by means of flow cytometry, western blot, real time-PCR and TransAMTM etc.in order to deeply discuss the role of Th1/Th2, Th17/Treg in RA. We also study the effects and regulatory mechanisms of dioscin of Rhizoma Dioscreae Nipponicae on the balance of Th1/Th2, Th17/Treg cells in the RA. This project can promote the understanding of RA immunological mechanisms in theory, and reveal the therapeutic mechanisms of dioscin of Rhizoma Dioscreae Nipponicae on RA. It provide a foundation for the choice of new effective therapeutic targets in RA and the further development and application of dioscin of Rhizoma Dioscreae Nipponicae.

CD4+T细胞在RA发病中起关键作用，其亚群在RA中的相互消长规律及与RA病程的关系尚未完全明了。本研究在CD4+T细胞亚群最新研究成果及去年底结题的国家基金项目基础上，拟以CIA鼠模型为研究对象，采用流式细胞术、western blot、实时荧光定量PCR、TransAMTM等方法，观察Thl/Th2、Th17/Treg及特异性转录因子、相关信号通路、相关细胞因子的变化规律，深入探讨Thl/Th2和Th17/Treg在RA中的作用及穿龙薯蓣皂苷对其的影响及调控机制。本项目的实施可以从理论上推动对RA的免疫学机制的阐释，揭示穿龙薯蓣皂苷治疗RA的机制，为治疗RA新的有效靶点的选择及穿山龙的进一步开发应用奠定基础。

CD4+T细胞在RA发病中起关键作用，但已有的研究报道在RA的病程的早、中、晚阶段Thl、Th17数量及相关细胞因子的表达存在矛盾。本课题组已有研究结果表明穿龙薯蓣皂苷具有很好的抗炎和免疫调节作用。本项研究以CIA小鼠为研究对象，通过体内外实验，采用流式细胞术、western blot、实时荧光定量PCR、细胞分选技术等方法，动态观察Thl/Th2、Th17/Treg及其相关细胞因子、信号转导子和转录激活因子、特异性转录因子等变化规律及与疾病严重程度的关系和穿龙薯蓣皂苷对其的影响。结果表明在CIA小鼠的病程中Thl细胞及其相关因子在CIA小鼠关节炎症早期起主要作用， Thl7细胞及其相关因子主要在中晚期逐渐发挥作用。CIA小鼠病程进展过程中Thl、Th17消长规律与CD4+T细胞相关的信号传导子和转录激活因子（STAT蛋白）及其特异性转录因子（T-bet、GATA3、RORγt、Foxp3）的活化及抑制有关。穿龙薯蓣皂苷能通过调节Thl、Th2、Th17、Treg信号转导子和转录激活因子及特异转录因子进而影响Thl、Th2、Th17、Treg的分化活化及相关细胞因子的分泌，从而起到对RA的有效治疗作用，初步阐明了穿龙薯蓣皂苷治疗RA的免疫学机制。