To compare different rat models of sepsis at different time points, based on pulmonary or extrapulmonary injury mechanisms, to identify a model which is more stable and reproducible to cause sepsis-associated acute lung injury (ALI). Adult male Sprague-Dawley rats were subjected to (1) cecal ligation and puncture (CLP) with single (CLP1 group) or two repeated through-and-through punctures (CLP2 group); (2) tail vein injection with lipopolysaccharide (LPS) of 10mg/kg (IV-LPS10 group) or 20mg/kg (IV-LPS20 group); (3) intratracheal instillation with LPS of 10mg/kg (IT-LPS10 group) or 20mg/kg (IT-LPS20 group). Each of the model groups had a sham group. 7-day survival rates of each group were observed (n=15 for each group). Moreover, three time points were set for additional experimental studying in each model group: 4 hours, 24 hours and 48 hours after modeling (every time point, n=8 for each group). Rats were sacrificed to collect BALF and lung tissue samples at different time points for detection of IL-6, TNF-alpha, total protein concentration in BALF and MPO activity, HMGB1 protein expression in lung tissues, as well as the histopathological changes of lung tissues. More than 50 % of the rats died within 7 days in each model group, except for the IT-LPS10 group. In contrast, the mortality rates in the two IV-LPS groups as well as the IT-LPS20 group were significantly higher than that in IT-LPS10 group. Rats received LPS by intratracheal instillation exhibited evident histopathological changes and inflammatory exudation in the lung, but there was no evidence of lung injury inCLP and IV-LPS groups. Rat model of intratracheal instillation with LPS proved to be a more stable and reproducible animal model to cause sepsis-associated ALI than the extrapulmonary models of sepsis.