Objective To investigate the role of heparanase (HPSE) in regulating the liver regeneration after major hepatectomy in rats. Methods The HPSE small interfering RNA (siRNA) was designed and synthesized, and was transfected into the rats after 70% hepatectomy by in vivo-jetPEI-Gal vector. Then immunohistochemistry was performed to determine the expression of hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF). Western blotting analysis was performed to detect the expression of matrix metalloprotein (MMP)-2 and MMP-9 protein expression. Immunostaining for Ⅷ factor was done to observe the microvessels, and the microvessel density (MVD) was calculated. Results Compared with control group, the expressions of HGF, VEGF, MMP-2 and MMP-9 were significantly inhibited in HPSE siRNA group, and the MVD was significantly decreased (P<0.01). Conclusion HPSE may regulate liver regeneration via promoting HGF release in ECM, inducing hepatocyte proliferation, increasing VEGF expression to accelerate angiogenesis and upregulating the expression of MMP-2 and MMP-9.