Objective To investigate the effect of different one-lung ventilation (OLV) time on TNF-α and IL-8 in rabbit bronchoalveolar lavage fluid, and the surfactant protein- A（SP-A） in lung tissue. Methods 60 male rabbits were randomly divided into group C, group O1, O2, O3, O4 and O5. Group O1-O5 were established the right lung ventilation and given 1, 2, 3, 4 and 5 hours ventilation. Five rabbits in each group were made execution after completing ventilation, the other five rabbits were executed after feeding 24 h. TNF-α, IL-8 concentrations and SA of all rabbits were measured. Results When compared with group C during OLV, cell number in left and right lung of group O3-O5 increased over 4.64±0.79（P＜0.01）, PaCO2 in group O4-O5 increased over 43.6±5.7 (P <0.01); TNF-α and IL-8 in left lung of group O3-O5 increased over 101.90±18.65（P＜0.01）, and TNF-α and IL-8 in right lung of group O4-O5 increased 256.16±30.32（P＜0.01）. 24 hours later, cell number in left lung was still higher than 7.5±1.9 (P <0.01); IL-8 in left lung of group O5 was still higher than 214.60±76.74（P＜0.01). TNF-α and IL-8 in left and right lung had no difference during OLV and 24 h later（P＞0.05）.When compared with group C, SP-A in left lung of group O1-O3 increased over than 13.84±3.47（P＜0.01）, and SP-A in right lung of group O1-O5 increased over than 5.51±1.48（P＜0.01）, but SP-A decreased expression in group O4-O5. SP-A in left and right lung of group O1-O5 increased expression more than 12.17±4.75 in 24 h later (P <0.01). Conclusions Lung inflammatory response increases and SP-A decreases with prolonged OLV; especially when OLV is longer than 3 h, lung injury is difficult to repair even after 24 h. It should be avoided to have long time one-lung ventilation.