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

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

Keywords:

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

PDF

___

Andersen, J. K. (2004). Oxidative stress in neurodegeneration: cause or consequence? Nature Medicine, 10(S7), S18–S25. https://doi.org/10.1038/nrn1434
Blesa, J., Trigo-Damas, I., Quiroga-Varela, A., and Jackson-Lewis, V. R. (2015). Oxidative stress and Parkinson’s disease. Frontiers in Neuroanatomy, 9. https://doi.org/10.3389/fnana.2015.00091
Coyle, J., and Puttfarcken, P. (1993). Oxidative stress, glutamate, and neurodegenerative disorders. Science, 262(5134), 689–695. https://doi.org/10.1126/science.7901908
Daiber, A., Mülsch, A., Hink, U., Mollnau, H., Warnholtz, A., Oelze, M., and Münzel, T. (2005). The Oxidative Stress Concept of Nitrate Tolerance and the Antioxidant Properties of Hydralazine. The American Journal of Cardiology, 96(7), 25–36. https://doi.org/10.1016/j.amjcard.2005.07.030
DiSabato, D. J., Quan, N., and Godbout, J. P. (2016). Neuroinflammation: the devil is in the details. Journal of Neurochemistry, 139, 136–153. https://doi.org/10.1111/jnc.13607
Dringen, R., Kussmaul, L., and Hamprecht, B. (1998). Detoxification of exogenous hydrogen peroxide and organic hydroperoxides by cultured astroglial cells assessed by microtiter plate assay. Brain Research Protocols, 2(3), 223–228. https://doi.org/10.1016/S1385-299X(97)00047-0
Dulce, R. A., Yiginer, O., Gonzalez, D. R., Goss, G., Feng, N., Zheng, M., and Hare, J. M. (2013). Hydralazine and Organic Nitrates Restore Impaired Excitation-Contraction Coupling by Reducing Calcium Leak Associated with Nitroso-Redox Imbalance. Journal of Biological Chemistry, 288(9), 6522–6533. https://doi.org/10.1074/jbc.M112.412130
Erel, O. (2004). A novel automated method to measure total antioxidant response against potent free radical reactions. Clinical Biochemistry, 37(2), 112–119. https://doi.org/10.1016/j.clinbiochem.2003.10.014
Erel, O. (2005). A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry, 38(12), 1103–1111. https://doi.org/10.1016/j.clinbiochem.2005.08.008
Forman, H. J. (2007). Use and abuse of exogenous H2O2 in studies of signal transduction. Free Radical Biology and Medicine, 42(7), 926–932. https://doi.org/10.1016/j.freeradbiomed.2007.01.011
Galvani, S., Coatrieux, C., Elbaz, M., Grazide, M.-H., Thiers, J.-C., Parini, A., … Nègre-Salvayre, A. (2008). Carbonyl scavenger and antiatherogenic effects of hydrazine derivatives. Free Radical Biology and Medicine, 45(10), 1457–1467. https://doi.org/10.1016/j.freeradbiomed.2008.08.026
Gandhi, S., and Abramov, A. Y. (2012). Mechanism of oxidative stress in neurodegeneration. Oxidative Medicine and Cellular Longevity, 2012, 428010. https://doi.org/10.1155/2012/428010
Hamann, K., Nehrt, G., Ouyang, H., Duerstock, B., and Shi, R. (2007). Hydralazine inhibits compression and acrolein-mediated injuries in ex vivo spinal cord. Journal of Neurochemistry, 071108171001006-??? https://doi.org/10.1111/j.1471-4159.2007.05002.x
Jessen, K. R. (2004). Glial cells. The International Journal of Biochemistry & Cell Biology, 36(10), 1861–1867. https://doi.org/10.1016/j.biocel.2004.02.023
Kaminskas, L. M., Pyke, S. M., and Burcham, P. C. (2004). Strong Protein Adduct Trapping Accompanies Abolition of Acrolein-Mediated Hepatotoxicity by Hydralazine in Mice. Journal of Pharmacology and Experimental Therapeutics, 310(3), 1003–1010. https://doi.org/10.1124/jpet.104.067330
Kruger, N. J. (1994). The Bradford Method for Protein Quantitation. In Protein Protocols Handbook, The (pp. 15–22). New Jersey: Humana Press. https://doi.org/10.1385/1-59259-169-8:15
Leiro, J. M., Álvarez, E., Arranz, J. A., Cano, E., and Orallo, F. (2004). Antioxidant activity and inhibitory effects of hydralazine on inducible NOS/COX-2 gene and protein expression in rat peritoneal macrophages. International Immunopharmacology, 4(2), 163–177. https://doi.org/10.1016/j.intimp.2003.10.004
Li, Y., Hou, D., Chen, X., Zhu, J., Zhang, R., Sun, W., … Kong, X. (2019). Hydralazine protects against renal ischemia-reperfusion injury in rats. European Journal of Pharmacology, 843, 199–209. https://doi.org/10.1016/j.ejphar.2018.11.015
Lu, T., Aron, L., Zullo, J., Pan, Y., Kim, H., Chen, Y., … Yankner, B. A. (2014). REST and stress resistance in ageing and Alzheimer’s disease. Nature, 507(7493), 448–454. https://doi.org/10.1038/nature13163
Magee, L. A. (2003). Hydralazine for treatment of severe hypertension in pregnancy: meta-analysis. BMJ, 327(7421), 955–0. https://doi.org/10.1136/bmj.327.7421.955
Maheshwari, M., Roberts, J. K., DeSutter, B., Duong, K. T., Tingling, J., Fawver, J. N., … Murray, I. V. J. (2010). Hydralazine Modifies Aβ Fibril Formation and Prevents Modification by Lipids in Vitro. Biochemistry, 49(49), 10371–10380. https://doi.org/10.1021/bi101249p
Mehrabani, M., Nematollahi, M. H., Tarzi, M. E., Juybari, K. B., Abolhassani, M., Sharifi, A. M., … Mirzamohammadi, S. (2020). Protective effect of hydralazine on a cellular model of Parkinson’s disease: a possible role of hypoxia-inducible factor (HIF)-1α. Biochemistry and Cell Biology, 98(3), 405–414. https://doi.org/10.1139/bcb-2019-0117
Ransohoff, R. M. (2016). How neuroinflammation contributes to neurodegeneration. Science (New York, N.Y.), 353(6301), 777–783. https://doi.org/10.1126/science.aag2590
Ray, P. D., Huang, B.-W., and Tsuji, Y. (2012). Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling. Cellular Signalling, 24(5), 981–990. https://doi.org/10.1016/j.cellsig.2012.01.008