Aim To investigate the protective effect of Salvianolate on the injury of human umbilical vein endothelial cells (HUVEC) induced by high glucose in vitro, and to explore its underlying mechanism. Method HUVEC were respectively cultured with 5.5 mmol/L glucose(Glu)(control group), 30 mmol/L Glu (high glucose injury group), 30 mmol/L Glu+50 mg/L Salvianolate (low dose Salvianolate group), 30 mmol/L Glu+100 mg/L Salvianolate (medium dose Salvianolate group), 30 mmol/L Glu+ 200 mg/L Salvianolate (high dose Salvianolate group) for 48 h. The morphous of cell was observed by inverted phase contrast microscope, and the cell viability as well as the activity of glutathione peroxidase (GSH-Px), malondialdehyde (MDA) content, the secretary content of nitric oxide(NO)and endothelin-1(ET-1), in supernatant were measured. Results In high glucose injury group, low dose Salvianolate group, medium dose Salvianolate group and high dose Salvianolate group, the cell viability, activity of GSH-Px and MDA content, secrete of ET-1 and NO were significantly different compared with those in control group(P<0.01); In low dose Salvianolate group, medium dose Salvianolate group and high dose Salvianolate group, the cell viability, activity of GSH-Px and MDA content, secrete of ET-1 and NO were significantly different compared with those in high glucose injury group(P<0.01). Conclusion The Salvianolate can protect HUVEC induced by high glucose in vitro, and the mechanism may be associated with scavenging reactive oxygen radicals, reducing lipid peroxidation, and increasing the activities of intracellular antioxidase, inhibiting the secrete of ET-1.