Objective To establish an animal model of transfusion-related acute lung injury (TRALI) and investigate the role of soluble CD40 ligand (sCD40L) in the development of TRALI. Methods The TRALI animal model established by trauma-hemorrhage-transfusion. Lung edema was evaluated by histopathological examination and the protein and Evans blue dye accumulation in bronchoalveolar lavage fluid. The concentration of sCD40L in storage packed red blood cell (PRBC) and rat's plasma was measured by enzyme-linked immunosorbent assay (ELISA). Results There were obvious epithelial hyperplasia and inflammatory cell infiltration in the lung tissue of 7 d-PRBC-treated group. The accumulation of protein in bronchoalveolar lavage fluid of 7 d-PRBC-treated group [(13.17±5.76)mg] was significantly higher than that in normal controls [(1.21±0.66)mg] and normal saline (NS)-treated group [(4.94±2.15) mg] (F=17.605,P<0.001). The leakage amount of Evans blue dye in 7 d-PRBC-treated group [(0.0109±0.0067)%/min] was significantly higher than that in NS-treated group [(0.0026±0.0006) %/min] (t=2.998,P=0.03). The concentration of sCD40L of the 7 d PRBC [(451.58±73.28) pg/ml] was significantly higher than 0 d PRBC [(277.94±98.18)pg/ml] (t=2.834,P=0.03). The concentration of sCD40L in the plasma of 7 d-PRBC-treated group [(878.21±125.30)pg/ml] was significantly higher than those in normal controls [(289.78±62.60)pg/ml] and NS-treated group [(418.07±47.68)pg/ml] (F=78.715,P<0.001). Conclusion The TRALI animal model was successfully established with trauma-hemorrhage-transfusion. The concentration of sCD40L in plasma of rats with massive transfusion is remarkably increased,suggesting sCD40L may play a role in the development of TRALI.