Abstract:
Objective To establish and assess the H
2O
2-induced oxidative stress model of Dami cells.
Methods Morphology of Giemsa-stained Dami cells was observed under microscope after they were treated with H
2O
2 at different concentrations. Proliferation of Dami cells was assayed with CCK-8 kit. Growth curve of Dami cells was plotted and 50% inhibiting concentration (IC
50) of H
2O
2 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 H
2O
2 concentration. The IC
50 of H
2O
2 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 H
2O
2. However, no expression of HO-1 and NQO1 was detected.
Conclusion Oxidative stress model of Dami cells can be established with H
2O
2 at the concentration of 1 mmol/L and anti-oxidative stress of Dami cells is associated with the Nrf2/ARE pathway.
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