Carvedilol protection against endogenous Aβ-induced neurotoxicity in N2a cells

Original Paper

Abstract

Mutations in amyloid precursor protein (APP) and presenilin1 result in overproduction and accumulation of β-amyloid (Aβ) peptide, which has been shown to play an important role in Alzheimer’s disease (AD) pathogenesis. Carvedilol, a nonselective β-adrenergic receptor blocker used for treatment for heart failure and hypertension, has displayed its neuroprotective capacity due to its antioxidant property. In this study, we investigated whether Carvedilol has a neuronal protective effect against endogenous Aβ neurotoxicity in mouse Neuro2a (N2a) cells transfected with Swedish amyloid precursor protein (Swe-APP) mutant and Presenilin exon9 deletion mutant (N2a/Swe.D9). Elevated levels of reactive oxygen species (ROS), protein carbonyls, and 4-HNE were found in N2a/Swe.D9 cells, which were ameliorated by administration of Carvedilol in a dose-dependent manner. In addition, the levels of ATP and mitochondrial membrane potential were reduced in N2a/Swe.D9 cells, which were restored by treatment with Carvedilol. N2a/Swe.D9 cells displayed increased vulnerability to H2O2-induced cell death and apoptosis, which could be attenuated by Carvedilol. Mechanistically, we found that Carvedilol prevented apoptosis signals through reducing cytochrome C release and the level of cleaved caspase-3. Taken together, our findings suggest a possible use of Carvedilol in AD treatment.

Keywords

Alzheimer’s disease Amyloid β Carvedilol Oxidative stress Apoptosis 

Supplementary material

12192_2018_881_Fig6_ESM.gif (114 kb)
Supplementary fig. 1

Effects of Carvedilol on the cell viability of N2a/Swe.D9 cells. N2a/Swe.D9 was treated with Carvedilol at the concentrations of 2 nm, 20 nM, 200 nM, 2 μM, 20 μM, 200 μM, and 2 mM for 24 h. Cell viability was determined by the MTT assay (*, P < 0.01; **, P < 0.0001 vs. untreatment group). (GIF 114 kb)

12192_2018_881_MOESM1_ESM.tif (59 kb)
High resolution image (TIFF 58 kb)
12192_2018_881_Fig7_ESM.gif (86 kb)
Supplementary fig. 2

Comparison of cell viability of N2a/Wt and N2a/Swe.D9 cells. Equal amount of cells (1 × 105) were plated in each well of 6-well plates. Cell viability was determined after 24 h and 48 h incubation by the MTT assay (*, P < 0.01 vs. N2a/Wt cells). (GIF 86 kb)

12192_2018_881_MOESM2_ESM.tif (51 kb)
High resolution image (TIFF 51 kb)

References

  1. Arrieta-Cruz I, Wang J, Pavlides C, Pasinetti GM (2010) Carvedilol reestablishes long-term potentiation in a mouse model of Alzheimer’s disease. J Alzheimers Dis 21(2):649–654.  https://doi.org/10.3233/JAD-2010-100225 CrossRefPubMedGoogle Scholar
  2. Butterfield DA, Drake J, Pocernich C, Castegna A (2001) Evidence of oxidative damage in Alzheimer’s disease brain: central role for amyloid β-peptide. Trends Mol Med 7(12):548–554.  https://doi.org/10.1016/S1471-4914(01)02173-6 CrossRefPubMedGoogle Scholar
  3. Carreira R, Duarte A, Monteiro P, Santos MS, Rego AC, Oliveira CR, Gonçalves LM, Providência LA (2004) Carvedilol protects ischemic cardiac mitochondria by preventing oxidative stress. Rev Port Cardiol 23(11):1447–1455PubMedGoogle Scholar
  4. Cotman CW, Su JH (1996) Mechanisms of neuronal death in Alzheimer’s disease. Brain Pathol 6(4):493–506.  https://doi.org/10.1111/j.1750-3639.1996.tb00878.x CrossRefPubMedGoogle Scholar
  5. Glat MJ, Offen D (2013) Cell and gene therapy in Alzheimer’s disease. Stem Cells Dev 22(10):1490–1496.  https://doi.org/10.1089/scd.2012.0633 CrossRefPubMedGoogle Scholar
  6. Green R, Reed JC (1998) Mitochondria and apoptosis. Science 281(5381):1309–1312.  https://doi.org/10.1126/science.281.5381.1309 CrossRefPubMedGoogle Scholar
  7. Hardy JA, Higgins GA (1992) Alzheimer’s disease: the amyloid cascade hypothesis. Science 256(5054):184–185.  https://doi.org/10.1126/science.1566067 CrossRefPubMedGoogle Scholar
  8. Hirai K, Aliev G, Nunomura A, Fujioka H, Russell RL, Atwood CS, Johnson AB et al (2001) Mitochondrial abnormalities in Alzheimer's disease. J Neurosci 21(9):3017–3023PubMedGoogle Scholar
  9. Kumar A, Prakash A, Dogra S (2011) Neuroprotective effect of Carvedilol against aluminium induced toxicity: possible behavioral and biochemical alterations in rats. Pharmacol Rep 63(4):915–923.  https://doi.org/10.1016/S1734-1140(11)70607-7 CrossRefPubMedGoogle Scholar
  10. LaFerla FM, Oddo S (2005) Alzheimer’s disease: Aβ, tau and synaptic dysfunction. Trends Mol Med 11(4):170–176.  https://doi.org/10.1016/j.molmed.2005.02.009 CrossRefPubMedGoogle Scholar
  11. Li-Sha G, Yi-He C, Na-Dan Z, Teng Z, Yue-Chun L (2013) Effects of Carvedilol treatment on cardiac cAMP response element binding protein expression and phosphorylation in acute coxsackievirus B3-induced myocarditis. BMC Cardiovasc Disord 13(1):100.  https://doi.org/10.1186/1471-2261-13-100 CrossRefPubMedPubMedCentralGoogle Scholar
  12. Lysko PG, Lysko KA, Webb CL, Feuerstein G (1992) Neuroprotective effects of Carvedilol, a new antihypertensive, at the N-methyl-D-aspartate receptor. Neurosci Lett 148(1–2):34–38.  https://doi.org/10.1016/0304-3940(92)90798-C CrossRefPubMedGoogle Scholar
  13. Mattson MP (2004) Pathways towards and away from Alzheimer’s disease. Nature 430(7000):631–639.  https://doi.org/10.1038/nature02621 CrossRefPubMedPubMedCentralGoogle Scholar
  14. Mohmmad Abdul H, Sultana R, Keller JN, St Clair DK, Markesbery WR, Butterfield DA (2006) Mutations in amyloid precursor protein and presenilin-1 genes increase the basal oxidative stress in murine neuronal cells and lead to increased sensitivity to oxidative stress mediated by amyloid β-peptide (1-42), H2O2 and kainic acid: implications for Alzheimer's disease. J Neurochem 96(5):1322–1335.  https://doi.org/10.1111/j.1471-4159.2005.03647.x CrossRefPubMedGoogle Scholar
  15. Oliveira PJ, Esteves T, Rolo AP, Palmeira CM, Moreno AJ (2004a) Carvedilol inhibits the mitochondrial permeability transition by an antioxidant mechanism. Cardiovasc Toxicol 4(1):11–20.  https://doi.org/10.1385/CT:4:1:11 CrossRefPubMedGoogle Scholar
  16. Oliveira PJ, Bjork JA, Santos MS, Leino RL, Froberg MK, Moreno AJ, Wallace KB (2004b) Carvedilol-mediated antioxidant protection against doxorubicin-induced cardiac mitochondrial toxicity. Toxicol Appl Pharmacol 200(2):159–168.  https://doi.org/10.1016/j.taap.2004.04.005 CrossRefPubMedGoogle Scholar
  17. Packer M, Bristow MR, Cohn JN, Colucci WS, Fowler MB, Gilbert EM, Shusterman NH (1996) The U.S. Carvedilol heart failure study group. The effect of Carvedilol on morbidity and mortality in patients with chronic heart failure. N Engl J Med 334(21):1349–1355.  https://doi.org/10.1056/NEJM199605233342101 CrossRefPubMedGoogle Scholar
  18. Reddy AP, Reddy PH (2017) Mitochondria-targeted molecules as potential drugs to treat patients with Alzheimer’s disease. Prog Mol Biol Transl Sci 146:173–201.  https://doi.org/10.1016/bs.pmbts.2016.12.010 CrossRefPubMedGoogle Scholar
  19. Rolo AP, Oliveira PJ, Moreno AJ, Palmeira CM (2003) Chenodeoxycholate induction of mitochondrial permeability transition pore is associated with increased membrane fluidity and cytochrome c release: protective role of Carvedilol. Mitochondrion 2(4):305–311.  https://doi.org/10.1016/S1567-7249(03)00007-2 CrossRefPubMedGoogle Scholar
  20. Rosenberg PB, Mielke MM, Tschanz J, Cook L, Corcoran C, Hayden KM, Norton M, Rabins PV, Green RC, Welsh-Bohmer KA, Breitner JC, Munger R, Lyketsos CG (2008) Effects of cardiovascular medications on rate of functional decline in Alzheimer disease. Am J Geriatr Psychiatry 16(11):883–892.  https://doi.org/10.1097/JGP.0b013e318181276a CrossRefPubMedPubMedCentralGoogle Scholar
  21. Selkoe DJ (1999) Translating cell biology into therapeutic advances in Alzheimer’s disease. Nature 399(6738 suppl):A23–A31.  https://doi.org/10.1038/399a023 CrossRefPubMedGoogle Scholar
  22. Sheng B, Gong K, Niu Y, Liu L, Yan Y, Lu G, Zhang L, Min H, Zhao N, Zhang X, Tang P, Gong Y (2009) Inhibition ofγ-secretase activity reduces Aβproduction, reduces oxidative stress increases mitochondrial activity and leads to reduced vulnerability to apoptosis: implications for the treatment of Alzheimer’s disease. Free Radic Biol Med 46(10):1362–1375.  https://doi.org/10.1016/j.freeradbiomed.2009.02.018 CrossRefPubMedGoogle Scholar
  23. Sherrington R, Rogaev EI, Liang Y, Rogaeva EA, Levesque G, Ikeda M, Chi H, Lin C, Li G, Holman K (1995) Cloning of a gene bearing missense mutations in early-onset familial Alzheimer’s disease. Nature 375(6534):754–760.  https://doi.org/10.1038/375754a0 CrossRefPubMedGoogle Scholar
  24. Smith MA, Harris PL, Sayre LM, Perry G (1997) Iron accumulation in Alzheimer’s disease is a source of redox-generated free radicals. Proc Natl Acad Sci U S A 94(18):9866–9868.  https://doi.org/10.1073/pnas.94.18.9866 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Strosznajder RP, Jesko H, Dziewulska J (2005) Effect of Carvedilol on neuronal survival and poly(ADP-ribose) polymerase activity in hippocampus after transient forebrain ischemia. Acta Neurobiol Exp (Wars) 65(2):137–143Google Scholar
  26. Wang L, Wang R, Jin M, Huang Y, Liu A, Qin J, Chen M, Wen S, Pi R, Shen W (2014) Carvedilol attenuates 6-hydroxydopamine-induced cell death in PC12 cells: involvement of Akt and Nrf2/ARE pathways. Neurochem Res 39(9):1733–1740.  https://doi.org/10.1007/s11064-014-1367-2 CrossRefPubMedGoogle Scholar
  27. Yamagata K, Ichinose S, Tagami M (2004) Amlodipine and Carvedilol prevent cytotoxicity in cortical neurons isolated from stroke-prone spontaneously hypertensive rats. Hypertens Res 27(4):271–282.  https://doi.org/10.1291/hypres.27.271 CrossRefPubMedGoogle Scholar
  28. Zuo L, Hemmelgarn BT, Chuang CC, Best TM (2015) The role of oxidative stress-induced epigenetic alterations in amyloid-β production in Alzheimer’s disease. Oxidative Med Cell Longev 2015:604658CrossRefGoogle Scholar

Copyright information

© Cell Stress Society International 2018

Authors and Affiliations

  1. 1.Department of NeurologyLiaocheng People’s HospitalLiaochengChina

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