The Protective Effect of Hydralazine against Hydrogen Peroxide (H2O2)-Induced Oxidative Damage in C6 Glial Cell Line

Abstract

Purpose: Recent studies have shown that hydralazine has positive effects on nervous system. However, its effect on hydrogen peroxide-induced oxidative damage in glial cells is still unclear. The current experiment was designed to examine the effect of hydralazine on glial damage after hydrogen peroxide-induced oxidative damage in C6 glial cells involving proinflammatory cytokines. Material and Methods: In this study, the C6 glioma cell line was used. Four cell groups were prepared to evaluate the effect of hydralazine on glial cell death after hydrogen peroxide-induced oxidative damage. The control group was without any treatment. Cells in the H2O2 group were treated with 0.5 mM H2O2 for 24 hours. Cells in the hydralazine group were treated with various concentrations (12.5, 25, 50, and 100 µM/mL) of hydralazine for 24 hours. Cells in the hydralazine + H2O2 group were pre-treated with various concentrations (12.5, 25 50, and 100 µM/mL) of hydralazine for 1 hour and then exposed to 0.5 mM H2O2 for 24 hours. After completing oxidative damage induction, the cell viability was evaluated XTT assay. Total antioxidant status (TAS), total oxidant status (TOS), tumor necrosis factor alpha (TNF-α), and interleukin-1 beta (IL-1 β) levels in the cells were measured by commercial kits. Results: Hydralazine at the concentrations of 50 and 100 µM/mL significantly increased the cell viability in C6 cells after hydrogen peroxide-induced oxidative damage (p < 0.001). It also significantly decreased the levels of TOS (p < 0.001) whereas rising TAS levels (p < 0.01) after hydrogen peroxide-induced oxidative damage. Moreover, hydralazine reduced TNF-α and IL-1 β levels in C6 cells after hydrogen peroxide-induced oxidative damage (P < 0.001). Conclusion: Hydralazine decreases glial cell death after hydrogen peroxide-induced oxidative damage in C6 cells by activating antioxidant system and inhibiting proinflammatory cytokines.

Keywords

• Hydralazine
• Oxidative Stress
• Cell Death
• Proinflammatory Cytokines
• C6 Glioma

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