Nox2 and Nox4 Participate in ROS-Induced Neuronal Apoptosis and Brain Injury During Ischemia-Reperfusion in Rats
Background: Previously studies have shown that Nox2 and Nox4, as members of nicotinamide adenine dinucleotide phosphate oxidase (NADPH oxidase, Nox), participate in brain damage caused by ischemia-reperfusion (I/R). The aim of this study is to investigate the effects of specific chemical inhibitors of Nox2 and Nox4 on cerebral I/R-induced brain injury in rats.
Methods: At 0.5 h before MCAO surgery, the rats were pretreated with vehicle, Nox2 inhibitor (gp91ds-tat), and Nox4 inhibitor (GKT137831), respectively. After reperfusion for 24 h, the infarct sizes of brain tissues in rats in various groups are determined. The penumbra (ischemic) tissues are collected to measure ROS levels, neuronal apoptosis, and degeneration, as well as the integrity of the blood-brain barrier (BBB) in brain tissues of rats.
Results: gp91ds-tat and GKT137831 pretreatment significantly reduced the infarct sizes in brain tissues of rats, effectively suppressed I/R-induced increase in ROS levels, neuronal apoptosis and degeneration, and obviously alleviated BBB damage.
Conclusion: Under cerebral I/R conditions, Nox2 inhibitor (gp91ds-tat) and Nox4 inhibitor (GKT137831) can effectively play a protective role in the brain tissues of rats.
KeywordsCerebral ischemia-reperfusion Nox2 Nox4 Middle cerebral artery occlusion Reactive oxygen species
Conflict of Interest: The authors declare that they have no conflict of interest.
- 4.Cao G, Ye X, Xu Y, Yin M, Chen H, Kou J, Yu B. YiQiFuMai powder injection ameliorates blood-brain barrier dysfunction and brain edema after focal cerebral ischemia-reperfusion injury in mice. Drug Des Devel Ther. 2016;10:315–25. https://doi.org/10.2147/DDDT.S96818.CrossRefPubMedPubMedCentralGoogle Scholar
- 7.De Silva TM, Brait VH, Drummond GR, Sobey CG, Miller AA. Nox2 oxidase activity accounts for the oxidative stress and vasomotor dysfunction in mouse cerebral arteries following ischemic stroke. PLoS One. 2011;6:e28393. https://doi.org/10.1371/journal.pone.0028393.CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Green DE, Murphy TC, Kang BY, Kleinhenz JM, Szyndralewiez C, Page P, Sutliff RL, Hart CM. The Nox4 inhibitor GKT137831 attenuates hypoxia-induced pulmonary vascular cell proliferation. Am J Respir Cell Mol Biol. 2012;47:718–26. https://doi.org/10.1165/rcmb.2011-0418OC.CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Green SP, Cairns B, Rae J, Errett-Baroncini C, Hongo JA, Erickson RW, Curnutte JT. Induction of gp91-phox, a component of the phagocyte NADPH oxidase, in microglial cells during central nervous system inflammation. J Cereb Blood Flow Metab. 2001;21:374–84. https://doi.org/10.1097/00004647-200104000-00006.CrossRefPubMedGoogle Scholar
- 12.Huttemann M, Lee I, Grossman LI, Doan JW, Sanderson TH. Phosphorylation of mammalian cytochrome c and cytochrome c oxidase in the regulation of cell destiny: respiration, apoptosis, and human disease. Adv Exp Med Biol. 2012;748:237–64. https://doi.org/10.1007/978-1-4614-3573-0_10.CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Kleinschnitz C, Grund H, Wingler K, Armitage ME, Jones E, Mittal M, Barit D, Schwarz T, Geis C, Kraft P, Barthel K, Schuhmann MK, Herrmann AM, Meuth SG, Stoll G, Meurer S, Schrewe A, Becker L, Gailus-Durner V, Fuchs H, Klopstock T, de Angelis MH, Jandeleit-Dahm K, Shah AM, Weissmann N, Schmidt HH. Post-stroke inhibition of induced NADPH oxidase type 4 prevents oxidative stress and neurodegeneration. PLoS Biol. 2010;8(9). https://doi.org/10.1371/journal.pbio.1000479.CrossRefGoogle Scholar
- 19.Kochanski R, Peng C, Higashida T, Geng X, Huttemann M, Guthikonda M, Ding Y. Neuroprotection conferred by post-ischemia ethanol therapy in experimental stroke: an inhibitory effect on hyperglycolysis and NADPH oxidase activation. J Neurochem. 2013;126:113–21. https://doi.org/10.1111/jnc.12169.CrossRefPubMedGoogle Scholar
- 21.Li H, Gao A, Feng D, Wang Y, Zhang L, Cui Y, Li B, Wang Z, Chen G. Evaluation of the protective potential of brain microvascular endothelial cell autophagy on blood-brain barrier integrity during experimental cerebral ischemia-reperfusion injury. Transl Stroke Res. 2014;5(5):618–26. https://doi.org/10.1007/s12975-014-0354-x.CrossRefPubMedGoogle Scholar
- 22.Li H, Wang Y, Feng D, Liu Y, Xu M, Gao A, Tian F, Zhang L, Cui Y, Wang Z, Chen G. Alterations in the time course of expression of the Nox family in the brain in a rat experimental cerebral ischemia and reperfusion model: effects of melatonin. J Pineal Res. 2014;57(1):110–9. https://doi.org/10.1111/jpi.12148.CrossRefPubMedGoogle Scholar
- 26.Pang T, Wang J, Benicky J, Sanchez-Lemus E, Saavedra JM. Telmisartan directly ameliorates the neuronal inflammatory response to IL-1beta partly through the JNK/c-Jun and NADPH oxidase pathways. J Neuroinflammation. 2012;9:102. https://doi.org/10.1186/1742-2094-9-102.CrossRefPubMedPubMedCentralGoogle Scholar
- 29.Shen J, et al. Interrupted reperfusion reduces the activation of NADPH oxidase after cerebral I/R injury. Free Radic Biol Med. 2011;50:1780–6. https://doi.org/10.1016/j.freeradbiomed.2011.03.028.CrossRefPubMedGoogle Scholar