Aim Lentiviral vectors targeting human lectin-like oxidized low density lipoprotein receptor-1 （LOX-1） gene with RNA interference （RNAi） were transfected into human umbilical vein endothelial cell （HUVEC）, and the protective mechanisms on the injury induced by oxidized low density lipoprotein （ox-LDL） were observed. Methods The LV-si-OLR1 optimal interference was selected from the small interfering RNA （siRNA） interference which validity had been verified and the virus titer was measured. HUVEC were transfected after 96 h. The expressions of mRNA and protein of LOX-1 were respectively detected by reverse transcription polymerase chain reaction （RT-PCR） and Western blot.72 h after transfection, HUVEC were cultured with ox-LDL, which final concentration was 150 mg/L. 24 h after cultured by ox-LDL, endothelial cells vigor were detected by methyl thiazolyl tetrazolium （MTT） method and the expression levels of nitric oxide （NO） were detected by nitrate reductase method the changes in the expression level of intercellular adhesion molecule-1 （ICAM-1）, monocyte chemotactic protein-1 （MCP-1）, RhoA, Rho kinase-1 （ROCK1）, Rho kinase-2 （ROCK2） were detected by Western blot in each group. Results The sequencing results confirmed that interference targeting human LOX-1 lentiviral vector was successfully constructed, which packaged lentiviral titer 6×10¹¹ TU/L. Compared the transfected with the non-transfected groups, the expression of LOX-1 mRNA and protein significantly decreased （P<0.01） Compared with the control group, ox-LDL treated group could decrease endothelial cells vigor and expression levels of NO, while increase the expression levels of ICAM-1, MCP-1, RhoA, ROCK1, ROCK2 （P<0.05） After suppressing the expression of LOX-1, compared with ox-LDL treated group, endothelial cells vigor and expression of NO were increased, while expressions of ICAM-1, MCP-1, RhoA, ROCK1, ROCK2 were restrained （P<0.05）. Conclusion RNA interference in the expression of LOX-1 could reduce endothelial cell injury by increasing endothelial cells vigor and expression of NO, while reducing expressions of ICAM-1, MCP-1 and the levels of Rho/Rho kinase activity, which provided experimental evidence for treating atherosclerosis for the use of targeting LOX-1 gene.