Abstract: OBJECTIVE  To explore the effect of Shenfu injection on vascular endothelial injury induced by oxidative stress and related signal transduction pathway.METHODS  Human umbilical vein endothelial cells ECV304 were cultured. The concentration of H₂O₂ for modeling and Shenfu injection for intervening were determined by methylthiazoletetrazolium (MTT) assay. The intracellular reactive oxygen species level was detected by DCFH-DA fluorescent probe. Cell apoptosis was measured by Hoechst staining and flow cytometry. Western blot was used to determine the expression of Cleaved Caspase-3 (C-Caspase-3), ERK, p-ERK, Bcl-2 and Bax proteins.RESULTS  H₂O₂ induced ECV304 vascular endothelial cell death in a dose-dependent manner. The content of ROS was promoted by H₂O₂, which could be antagonized by Shenfu injection. Compared with the control group, H₂O₂ could increase the apoptosis of ECV304 cell, promote the activation of ERK and Caspase-3, and decrease Bcl-2/Bax. Shenfu injection or NAC could reduce H₂O₂-induced ECV304 vascular endothelial cell apoptosis, inhibit the activation of ERK and Caspase-3, and up-regulate Bcl-2/Bax (P < 0.05).CONCLUSION  Shenfu injection can reduce H₂O₂-induced ECV304 cell apoptosis by decreasing the phosphorylation of ERK, activating Caspase-3, and up-regulating Bcl-2/Bax.