Luminex液相蛋白芯片用于研究着床窗期子宫内膜和早孕蜕膜基质细胞的细胞因子的表达谱

Expression of cytokines in endometrial stromal cell of implantation window and decidua stromal cell of early pregnancy by Luminex multiplex analysis

目的探索着床窗期子宫内膜和早孕期蜕膜基质细胞的细胞因子谱表达及其对胚胎着床和早期妊娠的可能影响。方法获取着床窗期子宫内膜基质细胞(ESC)和早孕期蜕膜基质细胞(DSC)进行体外培养,采用Luminex液相蛋白芯片技术检测细胞培养上清液中15种细胞因子:肿瘤坏死因子(TNF-α)、干扰素-γ(IFN-γ)、白细胞介素(IL)-1β、IL-2、IL-4、IL-6、IL-8、IL-10、IL-12、颗粒-巨噬细胞集落刺激因子(GM-CSF)、颗粒细胞集落刺激因子(G-CSF)、巨噬细胞炎症蛋白-1α(MIP-1α)、单核细胞趋化蛋白因子-1(MCP-1)、受调节激活的正常T细胞排泌的因子(RANTES)和γ干扰素诱导蛋白10(IP-10)的含量变化,比较两者表达的差异。结果DSC分泌多种细胞因子,与ESC相比,其中TNF-α、IFN-γ、IL-1β、IL-6、IL-8、IL-10、IP-10、GM-CSF、G-CSF、MIP-1α、RANTES表达下调,有极显著差异(P<0.001);IL-4表达上调,有显著差异(P<0.01);MCP-1、IL-2和IL-12表达无明显变化。不同时期表达的细胞因子之间存在复杂的相关关系。结论从着床窗期子宫内膜到早孕期蜕膜基质细胞表达的细胞因子谱的水平发生变化,可能对调控胚胎着床和维持早期妊娠起着重要作用。

Objective： To investigate the expression profile of cytokines in endometrial stromal cells of implantation window and decidual stromal cells of early pregnancy. Methods： Purified endometrial stromal cells （ESC） from human endometrium of implantation window and decidual stromal cells （DSC） from decidua of early pregnancy were cultured in vitro and the culture media were collected. The concentrations of 15 cytokines in the culture media were detected by the Luminex xMAP system. Results： The Levels of TNF-α, IFN-γ, IL-1β, IL-6, IL-8, IL-10, IP-10, GM-CSF, G-CSF, MIP-1α, RANTES in culture medium of DSC were significantly lower than those in the culture media of ESC, while IL-4 were significantly higher. The levels of MCP-1, IL-2, IL-12 did not show significant difference between DSC and ESC. There were complicated relationships among different kinds of cytokines. Conclusions： Some kinds of cytokines in the endometrium were up-regulated and some were down-regulated from stage of implantation window to the stage of early pregnancy. The temporal and cellular expression patterns of the cytokines in this transition indicate that the cytokines could be the key regulators of human implantation and pregnancy establishment.