Ventilator-induced lung injury (Vili) is a life-threatening condition caused by the inappropriate use of mechanical ventilation (MV). However, the precise molecular mechanism inducing the development of Vili remains to be elucidated. in the present study, it was revealed that the calcineurin/nFaTc4 signaling pathway mediates the expression of adhesion molecules and proinflammatory cytokines essential for the development of Vili. The present results revealed that a high tidal volume ventilation (HV) caused lung inflammation and edema in the alveolar walls and the infiltration of inflammatory cells. The calcineurin activity and protein expression in the lungs were increased in animals with Vili, and nFaTc4 translocated into the nucleus following calcineurin activation. Furthermore, the translocation of nFaTc4 and lung injury were prevented by a calcineurin inhibitor (csa). Thus, the present results highlighted the critical role of the calcineurin/nFaTc4 signaling pathway in Vili and suggest that this pathway coincides with the release of icaM-1, VcaM-1, TnF-α and il-1β.