Objective To investigate the influence of high mobility group box 1(HMGB1) on the expression of interleukin 6(IL-6), receptor activator of nuclear factor-kappa B ligand(RANKL) and osteoprotegerin(OPG) on periodontal ligament fibroblasts. Methods Human periodontal ligament fibroblasts were stimulated with HMGB1 at concentrations of 10, 30, and 100 ng·mL-1 for 24 h. RT-PCR and Western blot analysis were performed to check mRNA and protein expression of IL-6, RANKL and OPG on the cells. Results The ratio of RANKL/OPG was increased at both mRNA and protein level after HMGB1 stimulation at 10, 30, 100 ng·mL-1. Inflammatory cytokine IL-6 was upregulated by HMGB1 at the concentration of 100 ng·mL-1. Conclusion Increased ratio of RANKL/OPG and IL-6 on periodontal ligament fibroblasts suggests that HMGB1 might play a role in the pathogenesis and progression of periodontal disease.