Aim To investigate the effect of high mobility group box 1 (HMGB1) and its receptor on survival and cytokine levels of rat marrow mesenchymal stem cells. Methods Marrow mesenchymal stem cells were treated with HMGB1 at 25.0,0.0 and 100.0 μg/L for 4,8 and 72 hours. Cell proliferations were detected by MTT after different treatment. VEGF and bFGF were detected by ELISA. Expressions of receptor of advanced glycation end products (RAGE) and Toll-like receptor 4 (TLR4) in mesenchymal stem cells were detected by Western blot. Real-time PCR and Western blot were used to examine RAGE and TLR4 expression, which were to verify the efficiency of transfection with siRNA in marrow mesenchymal stem cells. MTT assay and ELISA were used to analyze proliferation and secretions of VEGF and bFGF in si-RAGE or si-TLR4 marrow mesenchymal stem cells. Results 25.0 μg/L HMGB1 could increase marrow mesenchymal stem cells proliferation (P＜0.05). 25.0 and 50.0 μg/L HMGB1 could promote VEGF and bFGF secretion compared with control group (P＜0.05). The protein levels of RAGE and TLR4 in 25.0 μg/L HMGB1 group were significantly higher than those in other groups (P＜0.05). Marrow mesenchymal stem cells transfected with si-TLR4 antagonized the cell proliferation and secretion of VEGF, bFGF induced by HMGB1. Conclusion HMGB1 promote cell proliferation and secret VEGF, bFGF by binding to its TLR4 receptor.