In response to danger signals, macrophages rapidly produce many inflammatory cytokines that trigger the cascade release of inflammatory mediators, leading to tissue damage, which is an important cause of clinical manifestations of syphilis at all stages. However, we still know very little about the specific mechanism of this process. Tp0768 is an infection-stage-dependent antigen that plays an important role in the infection of Treponema pallidum. In this study, we demonstrated that Tp0768 stimulation of macrophages can cause IL-1 beta, IL-6, and IL-8 mRNA expression levels to increase in a dose- and time-dependent manner. Further research showed that Tp0768 activated ER stress and the ROS/NF-kappa B pathway in macrophages. Inhibition of ER stress and the ROS/NF-kappa B pathway inhibited the expression of IL-1 beta, IL-6, and IL-8 induced by Tp0768. In addition, pretreatment with a PERK pathway inhibitor significantly reduced the expression of the NF-kappa B and JNK pathways, while also downregulating the expression of IL-1 beta, IL-6, and IL-8. Tp0768 stimulation can activate IRE1 alpha/XBP-1 signaling and participate in the induction of inflammatory cytokines through the JNK pathway. These findings indicate that Tp0768 promotes the secretion of proinflammatory cytokines IL-1 beta, IL-6, and IL-8 by macrophages through ER stress and the ROS/NF-kappa B pathway, which are also involved in the activation of the NF-kappa B and JNK pathways that are induced by the PERK pathway and activation of IRE1 alpha/XBP-1 signaling. Key points This study found for the first time that the recombinant Treponema pallidum protein Tp0768 promotes the production of IL-1 beta, IL-6, and IL-8 by macrophages through ER stress. Recombinant Treponema pallidum protein Tp0768 regulates the ROS/NF-kappa B pathway through ER stress. ER stress-related pathway PERK induces the expression of IL-1 beta, IL-6, and IL-8 by activating the NF-kappa B pathway and the JNK pathway. IRE1 alpha can induce the splicing of XBP-1mRNA and activate the JNK pathway.