Objective To explore the role of hydrogen peroxide (H2O2) in inducing chronic oxidative stress in microglia aging. Methods BV2 microglia purchased from ATCC in less than 10 generations were treated with 0, 50, 100, 200 μmol/L H2O2 at different concentrations. According to the concentration of H2O2 used, the BV2 microglia were divided into a control group and H2O2 -50 μmol/L Group, H2O2 -100 μmol/L Group, H2O2 -200 μmol/L Group. Cell proliferation was measured by CCK8 cell proliferation assay. Age-related β-galactosidase (SA-β-gal) staining assay, and expression of age-related cyclin molecules p16, p21, p53 and senescence sssociated secretory phenotype interleukin 1 beta (IL-1β), transforming growth factor-β (TGF-β) and matrix metalloprotein 9 (MMP9) detected by quantitative real-time polymerase chain reaction were used to measure celluar senescence. Results During the induction process, H2O2-200 μmol/L caused significant damage to BV2 microglia, therefore no subsequent testing was conducted. Finally, the control group, H2O2-50 μmol/L group and H2O2-100 μmol/L group cells were collected. The differences in cell survival rate (F=46.176, P<0.001) and positive rate of SA-β-gal staining (F=553.1, P<0.001) among the three groups were statistically significant. The cell survival rate of H2O2-50 μmol/L group had no significant change (P>0.05), while the cell survival rate of H2O2-100 μmol/L group decreased significantly (P<0.001). The positive rate of SA-β-gal staining in H2O2-50 μmol/L group and H2O2-100 μmol/L group was increased (P<0.001), and the positive rate of SA-β-gal staining in H2O2-100 μmol/L group was higher than that in H2O2-50 μmol/L group (P<0.001). The mRNA levels of senescence related cyclin molecules p16, p21 and p53 were up-regulated under the induction of 50, 100 μmol/L H2O2 (P<0.05), and the expressions of IL-1β, TGF-β and MMP9 of senescence associated secretory phenotype were increased (P<0.05). The increase of H2O2-50 μmol/L group was more obvious (P<0.05). Conclusion The aging model of BV2 microglia can be successfully established by inducing 8 d with 100 μmol/L H2O2, and the mechanism may be related to promoting the secretion of p16, p21, p53, IL-1β, TGF-β and MMP9.