Mycoplasma genitalium lipid-associated membrane proteins (LAMPg) can induce human monocytic cell line THP-1 to produce proinflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha), IL-1beta and IL-6, as demonstrated by an enzyme-linked immunosorbent assay and reverse transcription-PCR (RT-PCR). This study also investigated the signaling transduction pathways involved in the production of these cytokines. THP-1 cells were stimulated with LAMPg and then examined for the activation of MAPKs, such as SAPK/JNK, p38, extracellular signal-regulated kinase (ERK)1/2 and NF-kappaB and AP-1. Western blot clearly showed that stress-activated protein kinase (SAPK)/c-Jun-N-terminal kinase (JNK), p38 and ERK1/2 were activated in response to LAMPg, peaking at 30 min. SAPK/JNK-specific inhibitor SP600125 slightly suppressed IL-6 production although no evident effects were obtained for TNF-alpha and IL-1beta; ERK1/2 inhibitor PD98059 blocked both TNF-alpha and IL-1beta, but not IL-6 production. However, p38 inhibitor SB203580 abrogated TNF-alpha, IL-1beta and IL-6 production. The DNA-binding activity of NF-kappaB and AP-1 was also assessed by an electrophoretic mobility gel shift assay, and an NF-kappaB specific inhibitor, pyrrolidine dithiocarbamate, profoundly inhibited the synthesis and production of the proinflammatory cytokines. Based on these results, this study concludes that MAPKs, NF-kappaB and AP-1 may play important roles in the genital tract inflammatory reaction after mycoplasma infection.