Ischemia-reperfusion injury (IRI) is a major cause of lung dysfunction during cardiovascular surgery, heart transplantation and cardiopulmonary bypass procedures, and the inflammatory response, oxidative stress, and apoptosis play key and allegedly maladaptive roles in its pathogenesis. The aim of this study was to initially elucidate whether ozone induces oxidative preconditioning by activating nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and secondly to determine whether ozone oxidative preconditioning (OzoneOP) protects the lung from IRI by attenuating nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3)-mediated inflammation, enhancing the antioxidant activity of Nrf2 and inhibiting apoptosis. Rats treated with or without OzoneOP (2 ml containing 100 μg/kg/day) were subjected to 1 h of lung ischemia followed by 2 h of reperfusion for 10 days. Lung damage, antioxidant capacity, inflammation and apoptosis were evaluated and compared among different groups after reperfusion. OzoneOP significantly ameliorated changes in lung morphology and protected the lung from IRI by attenuating oxidative stress, inflammation-induced injury and lung apoptosis. Moreover, OzoneOP increased the expression of Nrf2 and decreased the levels of NLRP3, apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC), un-cleavable cysteine-requiring aspartate protease-1 (procaspase-1), cysteine-requiring aspartate protease-1 (caspase-1) and interleukin-1β (IL-1β) in the rat lungs. In summary, these results provide new insights into the molecular events modulated by ozone and suggest that ozone therapy may be an integrative support for patients with lung IRI.