内毒素血症Rig-Ⅰ表达调控的机制研究

Regulation mechanism for retinoic acid-inducible gene-Ⅰ in endotoxemia

[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 explore the regulation mechanism of retinoic acid-inducible gene-Ⅰ in endotoxemia. [Methods]（1） Binding proteins that interact with Rig-Ⅰ promoter in the nuclear extracts from liver between control and endotoxemic mice were screened,isolated and identified by DNA-pull down assay,2D difference gel electrophoresis（ 2D-DIGE） analysis and mass spectrum technology（ MS）;（2） hnRNP A3 mRNA expression and subcellular location of hnRNP A3 after LPS stimulation in RAW264. 7 cells were detected by quantitative real-time PCR and cell immunofluorescence respectively. [Results]（1） Seven differential proteins were screened and identified that binding with Rig-Ⅰ promoter in endotoxemia;（2） LPS induced hnRNP A3 mRNA expression（ P 〈0. 01）,and hnRNP A3 was found to be located in the nuclei. [Conclusion] Seven proteins were found to be involved in Rig-Ⅰ gene regulation in endotoxemia,which lay good foundation for further understanding the regulation mechanism of Rig-Ⅰ gene expression.