Advertisement

Anti-inflammatory Effect of AZD6244 on Acrolein-Induced Neuroinflammation

  • Wen-Chien Ho
  • Chia-Chi Hsu
  • Hui-Ju Huang
  • Hsiang-Tsui WangEmail author
  • Anya Maan-Yuh LinEmail author
Article

Abstract

Clinically, high levels of acrolein (a highly reactive α, β-unsaturated aldehyde) and acrolein adducts are detected in the brain of patients with CNS neurodegenerative diseases, including Alzheimer’s disease and spinal cord injury. Our previous study supports this notion by showing acrolein as a neurotoxin in a Parkinsonian animal model. In the present study, the effect of AZD6244 (an ATP non-competitive MEK1/2 inhibitor) on acrolein-induced neuroinflammation was investigated using BV-2 cells and primary cultured microglia. Our immunostaining study showed that lipopolysaccharide (LPS, an inflammation inducer as a positive control) increased co-localized immunoreactivities of phosphorylated ERK and ED-1 (a biomarker of activated microglia) in the treated BV-2 cells. Similar elevation in co-localized immunoreactivities of phosphorylated ERK and ED-1 was detected in the acrolein-treated BV-2 cells. Furthermore, Western blot assay showed increases in phosphorylated ERK in BV-2 cells subjected to LPS (1 μg/mL) or acrolein (30 μM); these increases were blocked by AZD6244 (10 μM). At the same time, AZD6244 attenuated LPS-induced TNF-α (a pro-inflammatory cytokine) and cyclooxygenase-II (COX II, a pro-inflammatory enzyme). Consistently, AZD6244 reduced acrolein-induced elevations in COX-II mRNA and COX-II protein expression. In addition, AZD6244 inhibited acrolein-induced increases in activated caspase 1 (a biomarker of inflammasome activation) and heme oxygenase-1 (a redox-regulated chaperone protein) in BV-2 cells. Using a transwell migration assay, AZD6244 attenuated acrolein (5 μM)-induced migration of BV-2 cells and primary cultured microglia. In conclusion, our study shows that acrolein is capable of inducing neuroinflammation which involved ERK activation in microglia. Furthermore, AZD6244 is capable of inhibiting acrolein-induced neuroinflammation. Our study suggests that ERK inhibition may be a neuroprotective target against acrolein-induced neuroinflammation in the CNS neurodegenerative diseases.

Keywords

Acrolein Neuroinflammation ERK pathway AZD6244 BV-2 cells 

Notes

Acknowledgments

The authors express their gratitude to Dr. C.Y. Chai at Institute of Biomedical Sciences, Academia Sinica, for his encouragement and support. Special thanks are due Dr. R.K. Freund at the Department of Pharmacology, University of Colorado, Anschutz, CO., USA, for editing this paper.

Funding Information

This study was financially supported MOST107-2320-B-010-019-MY3, MOST106-2320-B-010-004, and V106C-046, Taipei, Taiwan, R.O.C.

Compliance with Ethical Standards

The animals were supplied by the National Laboratory Animal Breeding and Research Center, Taipei, Taiwan, R.O.C. One animal was individually housed in an air-conditioned room (22 ± 2 °C) on a 12-h light/dark cycle (07:00–19:00 h light) and had free access to food and water. The use of animals has been approved by the Institutional Animal Care and Use Committee of Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C. The approval number is IACUC2017-242. All experiments were performed in accordance with relevant guidelines and regulation.

Conflict of Interest

The authors declare that they have no conflicts of interest.

References

  1. 1.
    Picklo MJ Sr, Montine TJ (2007) Mitochondrial effects of lipid-derived neurotoxins. J Alzheimer’s Dis : JAD 12(2):185–193CrossRefGoogle Scholar
  2. 2.
    Perluigi M, Coccia R, Butterfield DA (2012) 4-Hydroxy-2-nonenal, a reactive product of lipid peroxidation, and neurodegenerative diseases: a toxic combination illuminated by redox proteomics studies. Antioxid Redox Signal 17(11):1590–1609.  https://doi.org/10.1089/ars.2011.4406 CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Shi R, Page JC, Tully M (2015) Molecular mechanisms of acrolein-mediated myelin destruction in CNS trauma and disease. Free Radic Res 49(7):888–895.  https://doi.org/10.3109/10715762.2015.1021696 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Igarashi K, Yoshida M, Waragai M, Kashiwagi K (2015) Evaluation of dementia by acrolein, amyloid-beta and creatinine. Clin Chim Acta; Int J Clin Chem 450:56–63.  https://doi.org/10.1016/j.cca.2015.07.017 CrossRefGoogle Scholar
  5. 5.
    Liu-Snyder P, McNally H, Shi R, Borgens RB (2006) Acrolein-mediated mechanisms of neuronal death. J Neurosci Res 84(1):209–218.  https://doi.org/10.1002/jnr.20863 CrossRefPubMedGoogle Scholar
  6. 6.
    Jia Z, Hallur S, Zhu H, Li Y, Misra HP (2008) Potent upregulation of glutathione and NAD(P)H:quinone oxidoreductase 1 by alpha-lipoic acid in human neuroblastoma SH-SY5Y cells: protection against neurotoxicant-elicited cytotoxicity. Neurochem Res 33(5):790–800.  https://doi.org/10.1007/s11064-007-9496-5 CrossRefPubMedGoogle Scholar
  7. 7.
    Dong L, Zhou S, Yang X, Chen Q, He Y, Huang W (2013) Magnolol protects against oxidative stress-mediated neural cell damage by modulating mitochondrial dysfunction and PI3K/Akt signaling. J Mol Neurosci: MN 50(3):469–481.  https://doi.org/10.1007/s12031-013-9964-0 CrossRefPubMedGoogle Scholar
  8. 8.
    Takano K, Ogura M, Yoneda Y, Nakamura Y (2005) Oxidative metabolites are involved in polyamine-induced microglial cell death. Neuroscience 134(4):1123–1131.  https://doi.org/10.1016/j.neuroscience.2005.05.014 CrossRefPubMedGoogle Scholar
  9. 9.
    Jia Z, Zhu H, Li Y, Misra HP (2009) Cruciferous nutraceutical 3H-1,2-dithiole-3-thione protects human primary astrocytes against neurocytotoxicity elicited by MPTP, MPP(+), 6-OHDA, HNE and acrolein. Neurochem Res 34(11):1924–1934.  https://doi.org/10.1007/s11064-009-9978-8 CrossRefPubMedGoogle Scholar
  10. 10.
    Chen Z, Park J, Butler B, Acosta G, Vega-Alvarez S, Zheng L, Tang J, McCain R et al (2016) Mitigation of sensory and motor deficits by acrolein scavenger phenelzine in a rat model of spinal cord contusive injury. J Neurochem 138(2):328–338.  https://doi.org/10.1111/jnc.13639 CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Tian R, Shi R (2017) Dimercaprol is an acrolein scavenger that mitigates acrolein-mediated PC-12 cells toxicity and reduces acrolein in rat following spinal cord injury 141 (5):708–720. doi: https://doi.org/10.1111/jnc.14025 CrossRefGoogle Scholar
  12. 12.
    Dang TN, Arseneault M, Murthy V, Ramassamy C (2010) Potential role of acrolein in neurodegeneration and in Alzheimer’s disease. Curr Mol Pharmacol 3(2):66–78CrossRefGoogle Scholar
  13. 13.
    Walls MK, Race N, Zheng L, Vega-Alvarez SM, Acosta G, Park J, Shi R (2016) Structural and biochemical abnormalities in the absence of acute deficits in mild primary blast-induced head trauma. J Neurosurg 124(3):675–686.  https://doi.org/10.3171/2015.1.jns141571 CrossRefPubMedGoogle Scholar
  14. 14.
    Wang YT, Lin HC, Zhao WZ, Huang HJ, Lo YL, Wang HT, Lin AM (2017) Acrolein acts as a neurotoxin in the nigrostriatal dopaminergic system of rat: involvement of alpha-synuclein aggregation and programmed cell death. Sci Rep 7:45741.  https://doi.org/10.1038/srep45741 CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Luo J, Shi R (2004) Acrolein induces axolemmal disruption, oxidative stress, and mitochondrial impairment in spinal cord tissue. Neurochem Int 44(7):475–486CrossRefGoogle Scholar
  16. 16.
    Uchida K, Kanematsu M, Morimitsu Y, Osawa T, Noguchi N, Niki E (1998) Acrolein is a product of lipid peroxidation reaction. Formation of free acrolein and its conjugate with lysine residues in oxidized low density lipoproteins. J Biol Chem 273(26):16058–16066CrossRefGoogle Scholar
  17. 17.
    Li L, Jiang L, Geng C, Cao J, Zhong L (2008) The role of oxidative stress in acrolein-induced DNA damage in HepG2 cells. Free Radic Res 42(4):354–361.  https://doi.org/10.1080/10715760802008114 CrossRefPubMedGoogle Scholar
  18. 18.
    Lovell MA, Markesbery WR (2006) Amyloid beta peptide, 4-hydroxynonenal and apoptosis. Curr Alzheimer Res 3(4):359–364CrossRefGoogle Scholar
  19. 19.
    Huang Y, Jin M, Pi R, Zhang J, Chen M, Ouyang Y, Liu A, Chao X et al (2013) Protective effects of caffeic acid and caffeic acid phenethyl ester against acrolein-induced neurotoxicity in HT22 mouse hippocampal cells. Neurosci Lett 535:146–151.  https://doi.org/10.1016/j.neulet.2012.12.051 CrossRefPubMedGoogle Scholar
  20. 20.
    Zhao WZ, Wang HT, Huang HJ, Lo YL, Lin AM (2017) Neuroprotective effects of baicalein on acrolein-induced neurotoxicity in the nigrostriatal dopaminergic system of rat brain. Mol Neurobiol 55:130–137.  https://doi.org/10.1007/s12035-017-0725-x CrossRefGoogle Scholar
  21. 21.
    Tanel A, Averill-Bates DA (2007) Inhibition of acrolein-induced apoptosis by the antioxidant N-acetylcysteine. J Pharmacol Exp Ther 321(1):73–83.  https://doi.org/10.1124/jpet.106.114678 CrossRefPubMedGoogle Scholar
  22. 22.
    Doggui S, Belkacemi A, Paka GD, Perrotte M, Pi R, Ramassamy C (2013) Curcumin protects neuronal-like cells against acrolein by restoring Akt and redox signaling pathways. Mol Nutr Food Res 57(9):1660–1670.  https://doi.org/10.1002/mnfr.201300130 CrossRefPubMedGoogle Scholar
  23. 23.
    Huang MH, Lee JH, Chang YJ, Tsai HH, Lin YL, Lin AM, Yang JC (2013) MEK inhibitors reverse resistance in epidermal growth factor receptor mutation lung cancer cells with acquired resistance to gefitinib. Mol Oncol 7(1):112–120.  https://doi.org/10.1016/j.molonc.2012.09.002 CrossRefPubMedGoogle Scholar
  24. 24.
    Bernabe R, Patrao A, Carter L, Blackhall F, Dean E (2016) Selumetinib in the treatment of non-small-cell lung cancer. Future oncology (London, England) 12(22):2545–2560.  https://doi.org/10.2217/fon-2016-0132 CrossRefGoogle Scholar
  25. 25.
    Matsuoka Y, Yang J (2012) Selective inhibition of extracellular signal-regulated kinases 1/2 blocks nerve growth factor to brain-derived neurotrophic factor signaling and suppresses the development of and reverses already established pain behavior in rats. Neuroscience 206:224–236.  https://doi.org/10.1016/j.neuroscience.2012.01.002 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Alquezar C, Esteras N, de la Encarnacion A, Moreno F, Lopez de Munain A, Martin-Requero A (2015) Increasing progranulin levels and blockade of the ERK1/2 pathway: upstream and downstream strategies for the treatment of progranulin deficient frontotemporal dementia. Eur Neuropsychopharmacol :J Eur Coll Neuropsychopharmacol 25(3):386–403.  https://doi.org/10.1016/j.euroneuro.2014.12.007 CrossRefGoogle Scholar
  27. 27.
    Gross AM, McCully CM, Warren KE, Widemann BC (2017) Plasma and cerebrospinal fluid pharmacokinetics of selumetinib in non-human primates (NHP). J Clin Oncol 35(15_suppl):e14070–e14070.  https://doi.org/10.1200/JCO.2017.35.15_suppl.e14070 CrossRefGoogle Scholar
  28. 28.
    Shi R, Rickett T, Sun W (2011) Acrolein-mediated injury in nervous system trauma and diseases. Mol Nutr Food Res 55(9):1320–1331.  https://doi.org/10.1002/mnfr.201100217 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Catorce MN, Gevorkian G (2016) LPS-induced murine neuroinflammation model: main features and suitability for pre-clinical assessment of nutraceuticals. Curr Neuropharmacol 14(2):155–164CrossRefGoogle Scholar
  30. 30.
    Park J, Min JS, Kim B, Chae UB, Yun JW, Choi MS, Kong IK, Chang KT et al (2015) Mitochondrial ROS govern the LPS-induced pro-inflammatory response in microglia cells by regulating MAPK and NF-kappaB pathways. Neurosci Lett 584:191–196.  https://doi.org/10.1016/j.neulet.2014.10.016 CrossRefPubMedGoogle Scholar
  31. 31.
    Tanel A, Averill-Bates DA (2007) P38 and ERK mitogen-activated protein kinases mediate acrolein-induced apoptosis in Chinese hamster ovary cells. Cell Signal 19(5):968–977.  https://doi.org/10.1016/j.cellsig.2006.10.014 CrossRefPubMedGoogle Scholar
  32. 32.
    Wilkinson BL, Landreth GE (2006) The microglial NADPH oxidase complex as a source of oxidative stress in Alzheimer’s disease. J Neuroinflammation 3:30.  https://doi.org/10.1186/1742-2094-3-30 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute of PharmacologyNational Yang-Ming UniversityTaipeiTaiwan
  2. 2.Department of OncologyNational Taiwan University HospitalTaipeiTaiwan
  3. 3.Department of Medical ResearchTaipei Veterans General HospitalTaipeiTaiwan
  4. 4.Faculty of PharmacyNational Yang-Ming UniversityTaipeiTaiwan

Personalised recommendations