Fulminant hepatic failure (FHF) is a devastating clinical syndrome with extremely poor prognosis and high mortality. Therefore, better treatment is urgently needed. Polydatin (PD), a traditional anti-inflammatory drug, has been described to protect against liver injury induced by certain hepatotoxins. The present study investigated the protective effect of PD against lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced FHF in mice and the underlying mechanism. Mice were pretreated with an increasing dose of PD (10, 30, and 100 mg/kg), following LPS/D-GalN challenge. The liver injury was assessed biochemically and histologically. We found that PD exerted a protective effect on LPS/D-GalN-induced FHF as evidenced by reducing sera alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, diminishing liver histopathological injury, and lowering mortality in a dose-dependent manner. In addition, pretreatment mice with PD dose-dependently suppressed tumor necrosis factor-alpha (TNF-alpha) production, myeloperoxidase (MPO) activity, intercellular adhesion molecule-1 (ICAM-1) and endothelial cell adhesion molecule-1 (ECAM-1) expression, caspase-3 activation, and transcription factor nuclear factor-kappa B(NF-kB) activity induced by LPS. These results suggested that PD could effectively protect from LPS/D-GalN-induced FHF and the protective effect afforded by PD probably contributed to reduce TNF-alpha production via inhibiting NF-kB activation.