Evidence have shown that the exposure to environmental stressors, as organophosphate pesticides, could be associated with the development of neurogenerative diseases. Chlorpyrifos (CPF) is one of the most widely used pesticide around the world in agricultural and non-agricultural sectors due to its effectiveness. However, the indiscriminate use of CPF has been demonstrated to be poisonous to non-target species, including humans through exposure via dietary intake, inhalation, and dermal absorption, specially due to its capacity to cross the blood-brain barrier. In this sense, this study aimed to investigate the inflammatory responses of CPF exposure in murine microglial cells. BV-2 cells were exposed to different concentrations of CPF pesticide (0.3 – 300 μM) for 96h. Cell proliferation was measured by MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. Microglial activation was assessed using Iba-1 and CD11b antibody markers by flow cytometry and gene expression of pro-inflammatory genes was performed by real-time quantitative polymerase chain reaction (RT-PCR). All tests were performed in triplicate and the data obtained were statistically analyzed by one- or two-way ANOVA followed by Dunnett's multiple comparison test. Subchronic exposure to CPF at the concentration of 1 µM significantly increased cell proliferation whereas highest concentrations (30 - 300 µM) were cytotoxic. Besides, the lowest concentrations induced activation of microglial cells (1 and 3 µM), confirmed by Iba-1 and CD11b marking, and upregulated pro-inflammatory cytokines IL-1β and NLRP3 genes expression (3 µM) in BV-2 cells. Overall, data showed for the first time that the exposure to low levels of CPF triggers pro-inflammatory states in microglial cells. The massive generation of microglia-mediated inflammatory mediators could lead to neuronal loss; therefore, inflammation is considered as a contributor to the pathogenesis of neurodegenerative disorders. These results provide important information on the potential role of microglial activation in CPF-induced neuroinflammation.