Objective To investigate the effects of CpG oligodeoxynucleotide (CpG ODN) on the proliferation and migration of macrophages induced by lipopolysaccharide (LPS) and its mechanism. Methods In vitro inflammatory cell model was established by 1 mg/L LPS added into the culture medium of mouse RAW264.7 macrophages. CCK-8 assay was performed to assess the effect of CpG ODN (500 nmol/L) on the proliferation of macrophages induced by LPS; TranswellTM assay was used to measure the effect of CpG ODN (500 nmol/L) on the migration of macrophages induced by LPS. Western blot analysis was performed to detect the phosphorylation of p38 mitogen-activated protein kinase (p38MAPK), c-Jun N-terminal kinase (JNK), extracellular regulated protein kinases (ERK) and nuclear factor kappa-B subunits 65 (NF-κBp65). The specific inhibitors for p38MAPK (SB203580), JNK (SP600125), ERK (PD98059) and NF-κBp65 (BAY11-7082) were used to further explore the possible mechanism underlying the effects of CpG ODN. Real-time PCR was used to detect the role of CpG ODN in LPS-induced transcription of COX2 and MCP-1 in macrophages. Results CpG ODN synergistically enhanced the proliferation and migration of LPS-stimulated macrophages and promoted the transcription of COX2 and MCP-1. It also selectively enhanced the phosphorylation of JNK and ERK proteins in MAPK signaling pathway. To blockage JNK and ERK signaling pathways with its specific inhibitors dramatically inhibited the effects of CpG ODN. Conclusion CpG ODN can synergistically promote the proliferation and migration of LPS-stimulated macrophages through JNK and ERK pathway as well as the transcription of COX2 and MCP-1.