Paraquat (PQ) is an herbicide commonly used worldwide. This herbicide is known to alter the human and animal immune systems. Many reports indicated that PQ impacts immune cell viability and functions. However, the underlying mechanism critical is still unknown. Therefore, the aim of this study was to evaluate effects of PQ on free radical production, oxidative stress, cell death, and pro-inflammatory gene expression of murine bone marrow-derived macrophages (BMDMs) from female C57BL/6NJcl mice in vitro. BMDMs were incubated with PQ at 0, 200, 400 µM for 24 h. Intracellular reactive oxygen species (ROS) production, apoptosis, cell viability, nitric oxide, inducible nitric oxide synthase (iNOS), and IL-6 expression of murine BMDMs were measured. The results revealed that PQ treatments led to decrease the cell viability and induced apoptotic cell death in a dose-dependent manner. Additionally, PQ induced reactive oxygen species (ROS) generation. The mRNA expression level of pro-inflammatory mediator gene IL-6 and inducible nitric oxide synthase (iNOS) were elevated, while the level of lipid peroxides (MDA) production was unaltered by PQ treatment. Interestingly, PQ led to a decrease in nitric oxide production depends on its concentration. These phenomena indicated that PQ increased cellular ROS production which induced apoptosis, and the herbicide triggers production of iNOS and IL-6 in murine BMDMs.