Abstract: Glucocorticoid (GC) activates or inhibits the transcription of target genes mainly through the specific receptor-glucocorticoid receptor (GR) which involved in protein-protein interaction (PPI) and protein-DNA interaction (PDI) network. Meanwhile, the intervention of GC on Toll-like receptors (TLRs) signaling network largely depends on enhancing natural inhibitors of TLR pathways, the direct inhibition of the TLR-activated transcription factors and the required cofactors of cross-utilization of GR and TLR signaling pathways. In this article, the transcriptional regulation of GR, TLRs signaling networks and crosstalk of GR and TLRs signaling network were summarized based on the latest advances, and the GR-mediated mechanisms of down-regulation of TLR signaling pathways was reviewed in order to provide more valuable references on revealing the molecular mechanisms of the regulation of GC on mammalian cells and clinical application.