深度测序技术分析布鲁菌感染巨噬细胞RAW264.7早期转录组学变化

Deep Sequenceing-based Early Transcriptome Analysis of the RAW264.7 Cell with the Brucella Infection

本研究试图寻找参与布鲁菌胞内感染相关的宿主相关基因,为从感染宿主角度阐述布鲁菌的致病机制奠定基础。布鲁菌感染小鼠巨噬细胞后,利用数字基因表达谱技术筛选小鼠巨噬细胞感染布鲁菌16M株的差异表达基因,并利用荧光定量PCR对差异表达基因进行验证。差异表达基因经GO Term、KEGG分析,识别感染后显著富集的信号通路。在感染后4h,筛选出差异表达基因3 576个,其中58%的基因表现上调。并且NOD凋亡信号通路、溶酶体信号通路、NOD受体信号通路、FcγR-介导的吞噬通路、p53信号通路、内质网蛋白处理相关通路被显著富集。利用数字基因表达谱技术成功分析巨噬细胞感染布鲁菌后转录组学变化,为布鲁菌致病机制的逐步阐述奠定基础。

This experiment was designed to screen the genes of host involved Brucella intracellular infection and lay a foundation for elaborating the pathogenic mechanism. The murine macrophages were infected with Brucella melitensis 16M strains, and then the differently expressed genes of macrophages were screened with the digital gene expression profiling technology. The genes differently expressed were verified with the quantitative real-time PCR. Then the genes were analyzed with the GO Term and KEGG to screen the signals significantly enriched. There were 3 576 genes expressed significantly difference 4 hours post infection, approximately 58% genes were up-regulated. NOD-like receptor signaling pathway, lysosome pathway, Fc gamma R-mediated phagocytosis, p53 signaling pathway, apoptosis pathway and protein processing in the endoplasmic reticulum pathway were enriched. Transcriptomics profile of murine macrophages infected with Brucella melitensis 16M strain was successfully analyzed.