Objective To explore the effect of medicated serum of Sangleng and Eshu on the expression of vascular endothelial growth factor and vascular endothelial cell proliferation induced by VEGF in vitro. Methods Medicated serum of Sangleng and Eshu was used to culture human umbilical vein endothelial cell (HUVEC-1) induced by VEGF. The morphologic changes of HUVEC-1 were observed with phase contrast microscope, and cell proliferation was detected by MTT method, and the expression of vascular endothelial growth factor protein and mRNA in endothelial cells was detected by Western blotting and RT-PCR. Results The medicated serum of 5.0, 2.5 g·kg~ -1 ·d~ -1 Sangleng and Eshu could cause arrangement disorder in the normal umbilical vein endothelial cells. The medicated serum of 5.0 g·kg~ -1 ·d~ -1 Sangleng and Eshu (10%, 5%, 2.5%) and medicated serum of 2.5 g·kg~ -1 ·d~ -1 (10%) could inhibit vascular endothelial cell proliferation remarkably (P0.05), as compared with the blank serum of identical concentration. The medicated serum of 5 g·kg~ -1 ·d~ -1 and 2.5 g·kg~ -1 ·d~ -1 (5%) could reduce significantly the expression of vascular endothelial growth factor protein and mRNA of HUVEC-1. Conclusion The medicated serum of Sangleng and Eshu could inhibit vascular endothelial cell proliferation induced by VEGF in vitro, and its mechanism might be associated with inhibited expression of vascular endothelial growth factor.