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[目的]探讨内毒素血症Rig-Ⅰ表达调控的机制。[方法](1)DNA-pull down、2D-DIGE结合生物质谱技术筛选分离、鉴定对照和内毒素血症小鼠肝组织核蛋白与Rig-Ⅰ基因启动子结合的差异蛋白质;(2)q PCR和细胞免疫荧光检测LPS刺激RAW264.7细胞hnRNP A3 mRNA表达及细胞内定位。[结果](1)筛选鉴定得到hnRNP A3等7种与内毒素血症Rig-Ⅰ基因启动子结合的蛋白质;(2)LPS刺激下hnRNP A3 mRNA显著升高(P<0.01),且具有明显的细胞核定位。[结论]共有7种蛋白质参与内毒素血症Rig-Ⅰ基因表达调控,为进一步Rig-Ⅰ表达调控的研究奠定了良好基础。
[Objective] To investigate the mechanism of Rig-Ⅰ expression regulation in endotoxemia. [Methods] (1) DNA-pull down and 2D-DIGE combined with bio-mass spectrometry were used to screen and identify the differential proteins that bind to the Rig-Ⅰ gene promoter in the control and endotoxemia mouse liver tissues. (2) The mRNA and protein expression of hnRNP A3 in LPS-stimulated RAW264.7 cells were detected by PCR and immunofluorescence. [Results] (1) Screening identified seven kinds of hnRNP A3 and other endotoxemia Rig-Ⅰ gene promoter binding protein; (2) LPS stimulated hnRNP A3 mRNA was significantly increased (P <0.01), and with significant The nuclear localization. [Conclusion] A total of seven proteins were involved in the regulation of Rig-Ⅰ gene expression in endotoxemia, which laid a good foundation for the further study on the regulation of Rig-Ⅰ expression.