Objective To investigate the protecting mechanism of heat stress pretreatment on acute lung injury(ALI). Methods The oleic acid ALI mouse model was built to dynamically observe the binding capacity and the binding affinity of glucocorticoid receptor(GR),the levels of GR,heat shock protein 90(Hsp90)and Hsp70 before and after hyperthermic stress pretreatment. Results Heat stress pretreatment had significant protective effect on ALI.Western blotting showed insignificant changes of GR levels but progressive increase of level of Hsp70 and Hsp90.Heat stress pretreatment exerted insignificant effect on Bmax and Kd of GR,shown by radio ligand binding assay after ALI. Conclusion The protective effects of heat stress pretreatment on ALI of mouse may relate to its ability of keeping stable GR level and increasing levels of Hsp70 and Hsp90.