Aim To discuss the protective effect and mechanism of salidroside on homocysteine(Hcy）-induced endothelial cell injury. Methods Human umbilical vein endothelial cells(HUVEC)were cultured in different concentrations of Hcy, then the concentration of positive significance(1 mmol/L)was picked out. HUVEC were cultured with different concentrations of salidroside and Hcy concetration for 24 hours was chosen. Cell viability was assessed with MTT assay, the mRNA levels of Bip, CHOP were detected by real-time quantitative PCR(RT-PCR）. The protein levels of Bip, CHOP, phosphorylation of PERK, IRE1α were examined by Western blot. Results We observed that Hcy(0.5 mmol/L, 1 mmol/L, 2 mmol/L and 4 mmol/L)induced dysfunction of HUVEC, increased the mRNA and protein levels of Bip and CHOP(P<0.05）, and elevated PERK and IRE1α phosphorylation(P<0.05)in HUVEC. Salidroside attenuated the cell damage effects of Hcy on HUVEC in a dose-dependent manner. The cell viability of HUVEC were up-regulated by salidroside compared with Hcy treated group(P<0.05）. We also found that mRNA and protein levels of Bip and CHOP were down-regulated by salidroside(300 μmol/L)compared with Hcy treated group(P<0.05）, while PERK and IRE1α phosphorylation were increased(P<0.05)in HUVEC. Conclusions These findings suggested that salidroside could attenuate high Hcy induced injury in HUVEC, partly through inhibiting endoplasmic reticulum stress pathway.