柴胡多糖对空肠弯曲菌离体再刺激狼疮样小鼠脾细胞作用研究

Effect of Bupleurum polysaccharides on splenocytes from lupus like mouse re-challenged with Campylobacter jejuni-S_(131) in vitro

空肠弯曲菌(Campylobacter jejuni-S_(131),CJ-S_(131))免疫小鼠建立诱导型狼疮样模型。模型鼠脾细胞加入CJ-S_(131)再次刺激以建立离体炎症模型,同时离体给予小叶黑柴胡多糖(Bupleurum smithii var.parvifolium polysaccharides,BPs),研究BPs防治狼疮的作用机制。BALB/c小鼠随机分为正常对照组、佐剂对照组和模型组。在Day 0和Day 14,以CJ-S_(131)免疫小鼠建立狼疮样模型。Day 19取各组脾细胞,再分为空白组、单给药物组(BPs 5、10、20和40μg·m L^(-1))、CJ-S_(131)单刺激组和CJ-S_(131)刺激剂联合药物处理组。ELISA法检测培养48 h上清中总免疫球蛋白G(Ig G)、抗双链DNA(ds DNA)抗体、干扰素-γ(IFN-γ)、白细胞介素^(-1)0(IL^(-1)0)和IL-17的水平,MTT法测定药物对狼疮样小鼠脾细胞增殖活性的影响。结果显示,BPs显著地降低CJ-S_(131)离体再刺激引起的总Ig G、抗ds DNA抗体、IFN-γ和IL-10的高表达。BPs对高表达的IL-17水平和脾细胞活性无明显影响。研究表明,CJ-S_(131)离体再刺激狼疮样小鼠脾细胞可建立离体炎症模型。

Mice were immunized with Campylobacter jejuni-S131（CJ-S131） to establish the lupus-like model. Splenocytes from lupus like mice were challenged with CJ-S131 to induce inflammatory response in vitro. Bupleurum smithii var. parvifolium polysaccharides（BPs） was added in the inflammatory model to observe its underlying mechanisms of action on lupus. BALB/c mice were randomly divided into three groups including normal control group, adjuvant control group and lupus-like model. Mice were immunized on Day 0 and 14 with CJ-S131 to establish lupus-like syndrome, and sacrificed on Day 19. Splenocytes from each group were collected and divided into blank control group, BPs added group（BPs 5, 10, 20, 40 μg·m L^-1）, CJ-S131 stimulated group, and CJ-S131 plus BPs group. The levels of total Ig G, anti-ds DNA antibody, interferon-γ, interleukin-10（IL-10） and IL-17 were quantified by ELISA. The proliferation of splenocytes was determined in the MTT assay. BPs significantly suppressed the high levels of total Ig G, anti-ds DNA antibody, IFN-γ and IL-10 stimulated by CJ-S131 and had no significant effects on increased IL-17 secretion and splenocytes proliferation. The results suggest that re-stimulation of splenocytes with CJ-S131 could establish an inflammatory model in vitro. The effect of BPs on lupus might is related to its inhibition of the production of autoantibodies and associated cytokines.