Either estrogen or hypertonic saline (HTS) administration can decrease lung inflammation after ischemia-reperfusion. The present study investigated whether combined treatment with estrogen and HTS could provide further protection from mesenteric ischemia-reperfusion (MSIR) injury. Male C3H/HeOuJ mice were randomly segregated into the following groups: sham-operated (sham), vehicle treatment followed by MSIR (vehicle treated), estrogen treatment followed by MSIR (estrogen treated), HTS treatment and MSIR (HTS treated), and combined treatment of estrogen plus HTS and MSIR (combined treated). In MSIR, mice were subjected to mesenteric ischemia for 60 min followed by reperfusion for 30 min. The histology of the lung and the gut was obtained. Lung injury was evaluated by lung edema and myeloperoxidase (MPO) activity; lung protein expression of macrophage migration inhibitory factor (MIF), toll-like receptor-4 (TLR4), phosphorylated inhibitory κBα (p-IκBα), and inducible nitric oxide synthetase (iNOS) were assayed. Survival analysis was monitored after MSIR for 120 min. Compared with those in the sham-treated group, the lung water ratio, MPO activity, and expressions of MIF, TLR4, p-IκBα, and iNOS were significantly increased in the vehicle-treated group. Diminished MIF, TLR4, p-IκBα, and iNOS expressions were found in the estrogen-treated group, whereas suppression of p-IκBα and iNOS was found in the HTS-treated group. Treatment with estrogen or HTS decreased the lung water and MPO activity. The combined treatment of estrogen and HTS further significantly diminished p-IκBα and iNOS expression, lung edema, and MPO activity and improved the inflammation of the lung and the morphology of the gut in histology compared with the results of a single treatment of estrogen or HTS. Survival analysis showed significantly increased survival in the combination-treated group compared with survival in the HTS-treated group. In conclusion, compared with single-agent use, treatment combining estrogen and HTS further decreases lung p-IκBα and iNOS expressions, as well as the degree of lung injury. These effects may result in better rates of survival from MSIR injury.