[Abstract] Objective To evaluate the effect of Shen-mai injection on janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway in mice with lipopolysaccharide-induced lung injury. Methods Thirty adult male Wister rats were randomly divided into 3 groups :control group(group C), LPS-induced lung injury group(group ALI), LPS-induced lung injury + Shen-mai injection group(group SM). LPS 5 mg/kg was injected intraperitoneally in group ALI and group SM, and then Shen-mai injection 2 ml was injected intraperitoneally． The left lung was lavaged, and broncho-alveolar lavage fluid(BALF)was collected for determination of concentrations of protein，tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β) and intercellular adhesion molecule-1(ICAM-1). The lung tissues were removed for determination of the wet to dry lung weight(W/D) ratio, the mRNA expression level of IL-1β, TNF-α and ICAM-1. The pathological changes of the lungs were determinated by using light microscope and the lung injury scores were also determinated and immunohistochemistry was used to detected the protein expression level of p-JAK2, p-STAT3,. Results The W/D ratio and lung scores were significantly decreased in group SM (6.4±2.1),(7.2±2.9),compared with group ALI(8.2±2.6)、(12.1±3.2). The concentrations of IL-1β, TNF-α and ICAM-1 in BALF(53±10)、(94±19)、(187±36)ng/L were significantly decreased than that in group ALI (71±11)、(141±27)、(273±41)ng/L. The protein expression level of p-JAK2 and p-STAT3 (0.33±0.09)、(0.32±0.09) were significantly decreased than that in group ALI (0.45±0.03)、(0.45±0.05). Conclusion Shen-mai injection attenuates lipopolysaccharide-induced lung injury in rats possibly through inhibiting JAK2/STAT3 pathway.