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.