Objective To evaluate the mechanism of ceramide in aged platelets mediated by transfusion-related acute lung injury (TRALI). Methods The second hit model of TRALI was established in mice via lipopolysaccharide prestimulation, and platelets were transfused and stored for 1-5 days. ARC39 (a specific acid sphingomyelinase inhibitor) or platelets from acid sphingomyelinase-deficient mice were used to treat BALB/ c mice. Temporal changes in platelet sphingomyelin composition were analyzed. We examined the accumulation of neutrophils in the lungs of TRALI mice, as well as the function of the endothelial barrier and the histology of injured lung tissue. Results Platelets were collected and stored for 1~ 5 days from mice prestimulated by LPS, which caused characteristic lung injury, and the severity of lung injury increased with time. Ceramide in platelets accumulated and decreased during storage. Compared with the control group, transfusion of ARC39 preconditioned platelets or acid sphingomyelinase deficient platelets significantly alleviated lung injury. Conclusions Aging platelets can lead to TRALI in mice, but this damage can be reversed and treated by an acid sphingomyelinase inhibitor or specific knockout of the acid sphingomyelinase gene. Such intervention measures for sphingolipid formation are expected to be an effective strategy for increasing the storage safety and shelf life of blood products.