Objective: To investigate the therapeutic effect of the combination treatment of bone mesenchymal stem cells (BMSC) and hepatocyte growth factor (HGF) gene therapy on limb ischemia of rabbits. Methods: Ischemic model of hind limb was induced in the right hind limb of 32 New Zealand rabbits by ligation of the deep femoral artery and complete excision of the superficial femoral artery. After the model was established, the rabbits were equally randomized into empty plasmid treatment group (control group), BMSC treatment group (BMSC group), HGF gene therapy group (HGF group) and BMSC treatment plus HGF gene therapy group (BMSC+HGF group). All treatment regimens were administered by direct intramuscular injection into the operated limb. The collateral vessel count was performed using angiography 28 days after treatment, and the expression of CD31 and HGF in the muscle tissues surrounding the injection sites was detected by immunohistochemical staining and Western blot analysis respectively, 30 days after treatment. Results:The collateral vessel count showed no statistical difference in BMSC group or HGF group compared with control group, but it increased significantly in BMSC+HGF group compared with other 3 groups (P＜0.05 or P＜0.01). The immunohistochemical staining showed that the CD31 expression level of all the treated groups was significantly higher than that of the control group (P＜0.05 or P＜0.01), and CD31 expression had no significant difference between BMSC group and HGF group, but both of which were significantly lower than that of BMSC+HGF group (both P＜0.05). The Western blot demonstrated that the HGF expression level of all the treated groups was significantly higher than that of the control group (P＜0.05 or P＜0.01), and it was increased in the order of BMSC, HGF and BMSC+HGF group that all had statistical differences (all P＜0.05). Conclusions: The combination treatment with BMSC and HGF gene therapy has synergistic effect that can effectively promote collateral vessel development, and thereby improve the perfusion of the ischemic limb of rabbit.