Objective To investigate effect of toll-like receptor 4 (TLR4)/nuclear factor κB (NF-κB) p56 pathway on liver damage in children with rotavirus (RV) enteritis. Methods A total of fifty children with RV enteritis were selected as RV group, while another 50 children with RV negative enteritis during the same period were selected as control group. The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured by automatic biochemical analyzer. TLR4 mRNA and NF-κB mRNA expression in peripheral blood mononuclear cells (PMBC) were detected by polymerase chain reaction (PCR); the expression level of NF-κB p65 protein was detected by Western Blot. The levels of serum AST, ALT and the incidence of liver damage were compared between the two groups. TLR4 mRNA, NF-κB mRNA and NF-κB p65 protein expression in PMBC were compared between the two groups. The expression levels of TLR4 mRNA, NF-κB mRNA and NF-κB p65 protein in PMBC were compared between children with and without liver damage in RV group. The correlation between liver function and TLR4/NF-κB p65 pathway in children with RV enteritis was analyzed by Pearson correlation method. Results The levels of serum AST and ALT as well as the incidence of liver injury in the RV group were significantly higher than those in the control group (P < 0.05). The expressions of TLR4 mRNA, NF-κB mRNA and NF-κB p65 protein of PBMC in the RV group were significantly higher than those in the control group (P < 0.05). In the RV group, the expressions of TLR4 mRNA, NF-κB mRNA and NF-κB p65 protein of PBMC in children with liver damage were significantly higher than those in children without liver damage (P < 0.05). Serum AST and ALT were positively correlated with TLR4 mRNA, NF-κB mRNA and NF-κB p65 protein expression in children with RV enteritis (P < 0.05). Conclusion The incidence of liver damage is higher in children with RV enteritis, which is related to TLR4/NF-κB p65 pathway. Overactivation of TLR4/NF-κB p65 pathway may be one of the causes of liver damage in children with RV enteritis.