Aims Natural products have been used as potentially important sources of novel antibacterials in combating pathogenic Staphylococcus aureus isolates, amajor problemaround the world. In this study, we aimed to investigate the antibacterial effects of pinocembrin (PNCB) against Staph.aureus pneumonia in a murine model and its influence on the production of Staph.aureus -haemolysin (Hla). Methods and Results The in vitro activities of PNCB on -haemolysin production were determined using haemolysis, Western blot and real-time RT-PCR assays. The viability and cytotoxicity assays were performed to evaluate the influence of PNCB on -toxin-mediated injury of human alveolar epithelial cells. Moreover, through histopathologic analysis, we further determined the in vivo effects of PNCB on Staph.aureus pneumonia in a mouse model. In vitro, PNCB at low concentrations exhibited inhibitory activity against -haemolysin production and attenuated -haemolysin-mediated cell injury. Furthermore, the in vivo findings demonstrated that PNCB protected mice from Staph.aureus pneumonia. Conclusions We have provided new evidenceof the effects of PNCB, which suggest that PNCB attenuated -haemolysin-mediated cell injury and protected mice from Staph.aureus pneumonia. Significance and Impact of the Study The findings indicate that PNCB may be used as a basis foranti-Staphylococcus agent.