Objectives Senescence is involved in many age-related diseases, such as atherosclerosis, hypertension and diabetes. So how to delay or solve the problems caused by ageing has become a hot topic in the past decade. To explore the best stress-induced premature senescence (SIPS) model in vitro, we applied different dosages of H₂O₂ to the primary human umbilical vein endothelial cells (HUVECs).

Methods SIPS was induced by H₂O₂ with different concentration (0, 20, 40, 60, 80, 100 μM) in HUVECs_. Senescence related gene was evaluated with western blot or real-time PCR. Senescence-associated beta-galactosidase (SA-β-gal) staining was used to examine the phenotype of senescence. Apoptosis was analysed by flow cytometry.

Results The SA-β-gal staining result showed that the senescent cell was detected more significantly with H₂O₂≥40 μM compared with the control group. Also the mRNA of p21 was upregulated significantly with 40μM H₂O₂ and more. The PAI-1 protein was increased significantly with 60 μM H₂O₂ and more. In the apoptosis aspect, there was a significant increased number of apoptotic cells in 80 and 100 μM group.

Conclusions H₂O₂ can induce senescence of the primary HUVECs. In consideration of the its apoptotic effect, the 60 μM H₂O₂ could be the best concentration to induce senescence. Because of the high relevance with human, the primary HUVECs is a better candidate applied to explore the mechanism and the improved treatment of senescence in vitro.

Besides H₂O₂, there are some other ways to induce senescence, such as UV and gene knock down. Among these methods, H₂O₂ induced senescence via oxidative stress is the most relavent to the enviroment in the aged population.