【Objective】 To investigate the effect and mechanism of ginsenoside Rb1 attenuating human umbilical vein endothelial cells（HUVEC） senescence induced by high glucose through Sirt3/SOD2 pathway. 【Methods】 The senescence of HUVEC induced by high glucose （40 mmol/L）was assessed by senescence- associated β-galactosidase（SA-β-Gal）staining，and the expression of plasminogen activator inhibitor 1（PAI- 1）and P16. Annexin V- FITC /PI was performed to measure apoptotic effect. The expression of sirtuins 3（Sirt3）and superoxide dismutase 2（SOD2）was detected by western blot. Meanwhile，the level of intracellular malondialdehyde（MDA）and the activity of SOD2 were measured.【Results】 Treatment of HUVEC with high glucose for 24 hours induced premature senescence instead of apoptosis，as indicated by a larger proportion of the cells stained with SA-β-Gal and the up-regulated expression of PAI-1 and P16. Pretreatment of HUVEC with ginsenoside Rb1 （40 μ mol/L） could reverse endothelial cell senescence ，as indicated by the reduced SA-β-Gal positive cells and the down- regulated expression of PAI- 1 and P16. Furthermore，ginsenoside Rb1 pretreatment upregulated the protein expression of Sirt3 and SOD2，and eventually increased the activity of SOD2 and decreased the level of MDA.【Conclusion】 Ginsenoside Rb1 could antagonize high glucose-induced premature senescence of HUVEC via Sirt3/SOD2 signaling pathway.