Hydrogen peroxide-induced oxidative stress model of human megakaryocytic Dami cells and its assessment

Objective To establish and assess the H₂O₂-induced oxidative stress model of Dami cells. Methods Morphology of Giemsa-stained Dami cells was observed under microscope after they were treated with H₂O₂ at different concentrations. Proliferation of Dami cells was assayed with CCK-8 kit. Growth curve of Dami cells was plotted and 50% inhibiting concentration (IC₅₀) of H₂O₂ was calculated. ROS levels in Dami cells were measured by Flow cytometry after DCFH-DA probe was loaded. Nrf2, NQO1, HO-1, and γ-GCLC expressions were detected by Western blot. Results The shrinkage of Dami cells increased and their survival rate decreased with the increasing H₂O₂ concentration. The IC₅₀ of H₂O₂ was 0.9132 ± 0.144 mmol/L 2 h after the Dami cells were treated at the concentration of 0.1, 1, and 10 mmol/L, respectively. The positive oxidative stress rate of Dami cells was 12.11%, 20.91%, and 52.53%, respectively. The expression levels of Nrf2 and γ-GCLC were higher 24 h after the Dami cells were treated with H₂O₂. However, no expression of HO-1 and NQO1 was detected. Conclusion Oxidative stress model of Dami cells can be established with H₂O₂ at the concentration of 1 mmol/L and anti-oxidative stress of Dami cells is associated with the Nrf2/ARE pathway.