Objective：To investigate if lipopolysaccharide （LPS） combined with fungal glucan （curdlan） activates Nod-like receptor pyrin domain-containing protein（NLRP3） inflammasome，and the activation could be influenced by paeoniflorin（PF） in human bronchial epithelial cells（16HBE），as well as the underlying mechanism. Methods：16HBE cells were stimulated by LPS combined with curdlan to establish inflammation models. The expressions of NLRP3，caspase-1，and interleukin（IL）-1β mRNA were analyzed by RT-PCR. The level of IL-1β in supernatants was evaluated by ELISA. The activity of caspase-1 was measured using a caspase-1 activity kit. Reactive oxygen species（ROS） were detected by flow cytometry. The expressions of NLRP3，caspase-1，and IL-1β protein in cells were determined by Western blot. Results：The expressions of NLRP3，caspase-1，IL-1β mRNA and protein were up-regulated in 16HBE cells activated by LPS combined with curdlan. The activity of caspase-1 and the levels of IL-1β in supernatants，as well as the concentrations of intracellular ROS were significantly increased； these effects were inhibited effectively by PF，especially in high concentration，through reducing the production of intracellular ROS. Conclusion：NLRP3 inflammasome plays an important role in combined LPS and curdlan infection in 16HBE cells，and PF may possess a certain anti-fungal infection effect via inhibiting the activation of NLRP3 inflammasome.