Aim To investigate the inhibitory effect of berberine on the proliferation of human umbilical vein endothelial cell (HUVEC) induced by oxidized low density lipoprotein (ox-LDL) and the changes of related signaling pathways. Methods HUVEC was cultured in vitro, and the proliferation of HUVEC was stimulated by ox-LDL and intervened with berberine at different concentrations. The inhibitory effect of berberine on proliferation of HUVEC was detected by cell counting kit 8 and EdU incorporation experiment. The expressions of related genes and proteins and the changes of signaling pathway were analyzed by real-time quantitative PCR and Western blot. Results In a concentration dependent manner, berberine (1~50 mg/L) significantly inhibited the HUVEC proliferation induced by ox-LDL, and reduced the expressions of proliferating cell nuclear antigen (PCNA), nuclear factor κB (NF-κB) and lectin-like oxidized low density lipoprotein receptor-1 (LOX-1). These effects were associated with decreased phosphorylation levels of PI3K/Akt, ERK1/2 and p38MAPK signaling pathways. Conclusion Berberine inhibits the proliferation of HUVEC induced by ox-LDL through the down-regulation of PCNA, NF-κB and LOX-1 expressions. PI3K/Akt, ERK1/2 and p38MAPK signaling pathways are involved in this process.