Ischaemia–reperfusion (IR) injury is a major issue in cardiac transplantation. Inflammatory processes play a major role in myocardial IR injury. Lipocalin‐2 (Lcn2), which is also known as neutrophil gelatinase‐associated lipocalin, has multiple functions that include the regulation of cell death/survival, cell migration/invasion, cell differentiation and iron delivery. In our study, the hearts of C57BL/6 mice were flushed with and stored in cold Bretschneider solution for 8 h and then transplanted into a syngeneic recipient. We found that Lcn2 neutralization decreased the recruitment of neutrophils and macrophages. Troponin T (TnT) production, 24 h after myocardial IR injury, was reduced through anti‐Lcn2 antibody administration. The cardiac output at 60 mmHg of afterload pressure was significantly increased in hearts administrated with anti‐Lcn2 antibody administration (anti‐Lcn‐2: 58.9 ± 5.62 ml/min; control: 25.8 ± 4.1 ml/min; P < 0.05). Anti‐Lcn2 antibody treatment suppressed M1 marker (IL‐12, IL‐23 and iNOS) expression but increased M2 marker (IL‐10, Arg1 and Mrc1) expression. Furthermore, in our vitro and vivo experiments, we found that anti‐Lcn2 antibody treatment failed to induce M1‐related gene expression in response to LPS and that Lcn2 neutralization enhanced the expression of M2‐related genes following IL‐4 treatment. In conclusion, Lcn2 promotes M1 polarization, and Lcn2 neutralization attenuates cardiac IR injury.