To explore the protective mechanism of CTRP3 on lipopolysaccharide (LPS)-induced acute lung injury (ALI). Methods ALI model was established by injecting 3 mg/kg LPS into the trachea of mice, and the mice were randomly divided into 4 groups: control group, LPS group, LPS+LV-NC group and LPS+LV-CTRP3 group. The expression level of CTRP3 in lung tissues was detected by quantitative real-time polymerase chain reaction (qRT-PCR). HE staining was used to evaluate lung histology. The expressions of tumor necrosis factor (TNF) -α, interleukin (IL)-6 and IL-1β in bronchoalveolar lavage fluid (BALF) were determined by enzyma-linked immunosorbent assay (ELISA). The contents of SOD, CAT, GSH-Px and MDA were detected by the kit. Western blot was used to detect the expression of mechanism-related proteins. Results Compared with the control group, the expression of CTRP3 in ALI mice lung tissue was significantly decreased (P＜0.05). Compared with LPS group, overexpression of CTRP3 could alleviate pathological injury, significantly reduce the total inflammatory cells and neutrophils (P＜0.05), inhibit the release of inflammatory mediators and oxidative stress response (P＜0.05). Overexpression of CTRP3 activated silencing information modulator 1 (SIRT1) to regulate p65 phosphorylation and p53 acetylation (P＜0.05). Conclusion CTRP3 plays a protective role in ALI mice by regulating NF-κB/p53 signaling pathway mediated by SIRT1, which is a promising treatment strategy for ALI.