目的探讨原因不明复发性流产(unexplained recurrent spontaneous abortion,URSA)和先兆流产患者外周血单核细胞上PD-1和Tim-3的表达及其细胞因子的分泌情况,为先兆流产和URSA发病机制研究提供理论依据。方法随机选取29例健康早孕妇女作为正常早孕组(A组),37例仅有少量阴道出血的早孕妇女为先兆流产组(B组),22例URSA早孕患者为URSA组(C组);应用密度梯度离心法分离外周血单个核细胞,然后采用流式细胞分析技术检测3组孕妇外周血中CD14+单核细胞上Tim-3、PD-1的表达水平;ELISA法检测外周血中IL-4、IL-12、IFN-γ的含量。结果与正常早孕组相比,先兆流产组和URSA组外周血单核细胞上Tim-3、PD-1表达降低,外周血IL-4表达降低,URSA组低于先兆流产组(P〈0.05);IFN-γ、IL-12表达增高,URSA组高于先兆流产组(P〈0.05);相关性分析提示先兆流产组及URSA组外周血单核细胞上Tim-3与PD-1之间呈正相关;Tim-3、PD-1与IL-12呈负相关,IL-12与IL-4呈负相关,与IFN-γ呈正相关。结论外周血单核细胞上Tim-3、PD-1可能通过调节IL-12产生间接调节Th1/Th2平衡参与先兆流产和URSA的发生。
This study was designed to investigate the expression of PD-1 and Tim-3 on monocytes in peripheral blood and the secretion of relative cytokines, and to provide theoretical basis for evaluating the pathogenesis of threatened abortion and unexplained recurrent spontaneous abortion (URSA). Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral vein blood of 29 cases of normal early pregnancy (group A), 37 cases of threatened abortion (group B) and 22 cases of URSA (group C) with density gradient centrifugation. Flow cytometry was used to detect the frequency of Tim-3 and PD-1 expression on CD14^+ monoeytes in peripheral blood of the three groups; ELISA was used to detect the concentration of IL-4, IL-12 and IFN-γ in the peripheral blood of three groups. SPSS 17.0 analysis showed that compared with normal early pregnancy group, Tim-3 and PD-1 expressed on monocytes were down-regulated and the level of IL-4 was decreased in peripheral blood of threatened abortion group and URSA group, and those of URSA group were lower than those of threatened abortion group (P〈0.05); while the concentration of IFN-γ and IL-12 were up-regulated, and those of URSA group were higher than those of threatened abortion group (P〈0.05). The correlation analysis suggested that Tim-3 was positively correlated with PD-1, Tim-3 and PD-1 were negatively correlated with IL-12 (P〈0.05), IL-12 was negatively correlated correlated with IFN-γ with IL-4 but positively in threatened abortion and URSA groups. These findings suggest that PD-1 and Tim-3 on monocytes in peripheral blood can regulate Th1/Th2 balance by regulating IL-12 secretion and then participate in the occurrence of threatened abortion and URSA.