OBJECTIVE: To probe into the protective impact of remifentanil (Rem) on mechanical ventilation–induced lung injury (VILI) in ICR mice and its possible mechanism. STUDY DESIGN: A VILI model of ICR mice was constructed through 40 mL/kg tidal volume ventilation. The impact of Rem at 0.08 mg/kg and 0.04 mg/kg on survival, arterial oxygen saturation, and wet/dry (W/D) ratio of lung tissues as well as total protein level in bronchoalveolar alveolar lavage (BAL) fluid were investigated. HE staining and lactate dehydrogenase (LDH) content measurement were applied to detect the protective effects of Rem on VILI. Western blot was performed to detect the impact of Rem on apoptosis-related proteins as well as JAK1/STAT3 pathway in lung tissues. Immunohistochemistry was conducted to detect the impact of Rem on p-JAK1 in lung tissues, and ELISA was applied to detect MPO, TNF-α, IL-1β, and IL-6 in BAL fluid. SOD, CAT, GSH, GPx, and GST levels, which are related to anti-oxidative stress, were detected in lung tissues. RESULTS: Rem signally improved the survival rate and arterial oxygen saturation and reduced the W/D ratio, total protein, and LDH levels in BAL fluid of lung tissues. Rem visually improved VILI-induced morphological deterioration, inhibited apoptosis-related protein expressions, and reduced inflammatory factor expression level (TNF-α, IL-1β, IL-6, IL8). Rem observably promoted SOD, CAT, GSH, GPx, and GST expression levels. The analysis on PPI network pointed out that JAK1/STAT3 was the key molecule of the network, and Rem might affect the JAK1/STAT3 pathway through influencing the changes in PTPN2 and SOCS3 protein expression. Rem signally inhibited the activation of JAK1/STAT3 pathway, downregulated SOCS3 protein expression, and upregulated the PTPN2 protein expression level. CONCLUSION: Rem is available to exert protective effects on VILI through inhibiting pulmonary edema, inflammatory factor expression, lung tissue apoptosis level, and oxidative stress, and its protective effects on VILI may be related to the JAK1/STAT3 pathway. (Anal Quant Cytopathol Histpathol 2021;43:525–536) © Science Printers and Publishers, Inc.