灵仙新苷上调胶原性关节炎大鼠肠道相关淋巴组织调节性T细胞发挥免疫抑制作用的体外实验研究

Objectives: To explore the immunosuppressive effects of Clematichinenoside AR (AR) involving regulatory T cells (Treg cells) derived from gut associated lymphoid tissues (GALT) of rats with collagen induced arthritis (CIA) in vitro. Methods: After the CIA model rats being established, lymphocytes derived from Peyer's patches (PPs) and mesenteric lymph nodes (MLNs) were isolated and cultured under sterile conditions, and cells were incubated with various concentrations of AR (1 μM, 10 μM, 100 μM) in vitro. Levels of IL-10 and TGF-β1 were measured by ELISA, lymphocyte proliferation was detected using Cell Counting Kit-8 (CCK-8), percentages of CD4+CD25+Foxp3+ T regulatory cells were determined by flow cytometry, and expression levels of Foxp3 protein were investigated by western blot analysis. Results: In vitro treatment with AR markedly increased the levels of IL-10 and TGF-β1 secreted from ConA-activated PPs lymphocytes obtained from CIA rats. Furthermore, AR treatment inhibited ConA-activated T cell proliferation, up-regulated the percentages of CD4+ CD25+ Foxp3+ Treg cells among CD4+ T cells and the expression levels of Foxp3 protein in ConA-activated MLNs lymphocytes obtained from CIA rats significantly. Conclusions: In conclusion, these results suggested AR might exert its immunosuppressive effects by up-regulating CD4+CD25+Foxp3+ Treg cells in GALT obtained from CIA rats in vitro.