Objective To investigate the effect of heme oxygenase‑1 (HO‑1) on the expression and nuclear translocation of bHLH leucine zipper transcription factor E3 (TFE3) expression, and golgi stress response in mice with endotoxin‑induced acute lung injury (ALI). Methods According to the random number table method, 24 mice were divided into four groups (n=6): a blank control (Ctrl) group, an endotoxin ［lipopolysaccharide (LPS)］‑induced acute lung injury (LPS) group, an endotoxin‑induced acute lung injury+HO‑1 agonist hemin (LPS+Hemin) group, and a Hemin group. Mice in the Ctrl group were injected with 0.5 ml of normal saline via the tail vein. Those in the LPS group were intravenously injected with LPS at 10 mg/kg via the tail vein. The LPS+Hemin group was intraperitoneally injected with hemin at 50 mg/kg, followed by intravenous injection of LPS at 10 mg/kg via the tail vein 1 h later to establish an ALI model. The Hemin group was intraperitoneally injected with hemin at 50 mg/kg. Then, 12 h after modeling, the mice were sacrificed and the lung tissues were harvested. Hematoxylin‑eosin (H‑E) staining was used to observe the histopathological changes of lung tissues and the pathological changes of lung tissues were scored. The lung wet to dry weight ratio (W/D ratio) was calculated. Cell apoptosis index was analyzed by terminal deoxynucleotidyl transferase‑mediated dUTP‑biotin nick end labeling (TUNEL). The levels of interleukin (IL)‑6 and tumor necrosis factor (TNF)‑α in lung tissues were determined by enzyme‑linked immunosorbent assay (ELISA). The content of reactive oxygen species (ROS) was detected by flow cytometry. TFE3 nuclear translocation was observed by immunofluorescent staining. The levels of HO‑1, TFE3, Golgi matrix protein130 (GM130), Golgi reassembly‑stacking protein of 65 kDa (GRASP65), vesicular transportor‑rab GTPase20 (RAB20), syntaxin3A (STX3A), and WD repeat domain phosphoinositide‑interacting protein1 (WIPI1) were measured by Western blot. Results Compared with the Ctrl group, the LPS group showed aggravated lung injury, and increases in lung injury scores, W/D values, cell apoptosis index, the contents of ROS, the concentrations of IL‑6 and TNF‑α, TFE3 expression and nuclear translocation, and the levels of HO‑1, GRASP65, RAB20 and STX3A (all P<0.05), as well as decreases in the levels of WIPI1 and GM130 (all P<0.05). No statistical differences were found as to the above‑mentioned parameters in the Hemin group (all P>0.05). Compared with the LPS group, the LPS+Hemin group presented relieved lung injury, decreases in lung injury scores and W/D values, cell apoptosis index, the contents of ROS, the concentrations of IL‑6 and TNF‑α, TFE3 expression and nuclear translocation, and the levels of HO‑1, WIPI1 and GM130 (all P<0.05), as well decreases in the levels of GRASP65, RAB20 and STX3A (all P<0.05). Conclusions HO‑1 attenuates endotoxin‑induced ALI in mice, which be related to its regulation of TFE3 expression, nuclear translocation and Golgi stress response.