Inhaled zinc oxide nanoparticles (ZnO-NPs) induce lung inflammation associated with oxidative stress. The NLRP3 inflammasome plays a pivotal role in the development of lung inflammation. However, the underlying effects of the NLRP3 inflammasome on ZnO-NPs-induced inflammation remain obscure. In the present study, reactive oxygen species (ROS) generation, expression of NLRP3, caspase-1 p10, and cytokines release of interleukin (IL)-1β and IL-18 were determined after A549 cells were exposed to ZnO-NPs. The ROS scavenger N-acetyl-L-cysteine (NAC), nuclear factor kappa B (NF-κB inhibitor BAY11-7082, and NLRP3 inhibitor glibenclamide (GEL) were used to explore the mechanism of NLRP3 inflammasome activation-induced by ZnO-NPs. ZnO-NPs stimulation induced ROS generation and NF-κB p65 phosphorylation. Similarly, the expression of NLRP3 and caspase-1 p10 and the release of IL-1β and IL-18 were significantly increased after ZnO-NPs treatment, which indicated that the NLRP3 inflammasome was activated by ZnO-NPs. Meanwhile, NAC pretreatment inhibited ZnO-NPs-induced activation of NF-κB and NLRP3 inflammasome. The NF-κB inhibitor BAY11-7082 did not affect ROS production but significantly reduced the NLRP3 inflammasome activation induced by ZnO-NPs. Furthermore, the ability of ZnO-NPs to increase the production of IL-1β and IL-18 was significantly inhibited by GEL. The ZnO-NPs induced the activation of the NLRP3 inflammasome in A549 cells, which might be via a ROS-NF-κB-NLRP3 signaling pathway.