Aim: The aim of the present study was to investigate the protective effects of vitamin D3 on LPS-induced liver injury in mice and the mechanisms of this effect. Methods: Forty C57BL/6 mice were randomly assigned to 4 groups (n = 10) as follows: control group, vitamin D3 group (Vit D3), model group (LPS), and treatment group (LPS + Vit D3). Acute liver injury of mice in model group and treatment group was induced by the intraperitoneal injection of 15 mg/kg LPS. Mice in the vitamin D3 and treatment groups were given 2.5μg/kg vitamin D3 at the time points of 0h, 8h, 16h after LPS injection, while mice in the control and model groups were treated with an equivalent volume of 0.9% sodium chloride solution. After 24 h, all mice were anesthetized with chloral hydrate. Blood and livers of mice were collected for subsequent experiments. Results：Vitamin D3 decreased the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum and liver MDA, increased the activity of antioxidant enzymes in the liver tissues compared with those in the model group and attenuated liver pathologic changes. In addition, vitamin D3 downregulated the serum levels of tumor necrosis factor-α(TNF-α), interleukin -1β(IL-1β) compared with those in the model group. Meanwhile, vitamin D3 downregulated the protein expression of IL-1β, TNF-α, NF-κB p65 and NF-κB p50，upregulated the expression of Nrf2, HO-1 and VDR in liver tissues compared with those in the model group. Conclusions: Vitamin D3 showed a protective effect against LPS-induced acute liver injury in mice and that this effect may be partly due to?the inhibition of oxidative stress and inflammation via the VDR pathway.