Journal of Molecular Neuroscience

, Volume 67, Issue 4, pp 495–503 | Cite as

Prestimulation of Microglia Through TLR4 Pathway Promotes Interferon Beta Expression in a Rat Model of Alzheimer’s Disease

  • Niloufar Yousefi
  • Fattah Sotoodehnejadnematalahi
  • Nooshin Heshmati-Fakhr
  • Mohammad Sayyah
  • Masoud Hoseini
  • Soheil Ghassemi
  • Shayan Aliakbari
  • Hamid Gholami PourbadieEmail author


Soluble amyloid beta (Aβ) oligomers are the most common forms of Aβ in the early stage of Alzheimer’s disease (AD). They are highly toxic to the neurons but their capability to activate microglia remains controversial. Microglia develop two distinct phenotypes, classic (M1) and alternative (M2). Tuning of microglia to the alternative (anti-inflammatory) state is of major interest in treatment of neuroinflammatory disease. This study aimed to assess tuning the microglia to produce interferon beta (IFN-β) as an anti-inflammatory cytokine through TLR4 pathway in a rat model of AD. Microglial BV-2 cells were treated with 1 μg/ml lipopolysaccharides (LPS), Monophosphoryl lipid A (MPL), or vehicles for 24 h, and then incubated with Aβ oligomer. After 24 h, cell pellets were harvested and TIR-domain-containing adapter-inducing interferon-β (TRIF), interferon regulatory factor 3 (IRF3), and IFN-β levels were measured. The ligands/vehicle were microinjected into the right ventricle of male Wistar rats every 3 days. Two weeks later, an osmotic pump filled with oligomeric Aβ/vehicle was implanted in the left ventricle. After 2 weeks, TRIF, IRF3, and IFN-β levels were measured in the hippocampal tissue. TNF-α and IFN-β levels were assessed in the hippocampus using immunohistochemistry. The oligomeric Aβ did not change TRIF, IRF3, and IFN-β levels in both cell culture and hippocampal tissue. However, pretreatment with LPS or MPL increased the level of these proteins. BV-2 cells morphologically express M1 state in presence of higher dose of Aβ oligomer (10 μM). Pretreatment with LPS or MPL decreased the TNF-α and increased the number of IFN-β positive cells in the hippocampus of Aβ-treated rats. In conclusion, pretreatment with low dose TLR4 agonists could induce microglia to produce neuroprotective cytokines including IFN-β which may be considered as a potential strategy to combat neuronal degeneration in AD.


Alzheimer’s disease MPL LPS TLR4 Interferon-β Microglia Beta amyloid 


Compliance with Ethical Standards

Conflicts of interest

The authors declare that there are no conflicts of interest.


  1. Aloisi F (2001) Immune function of microglia. Glia 36:165–179CrossRefGoogle Scholar
  2. Buchanan MM, Hutchinson M, Watkins LR, Yin H (2010) Toll-like receptor 4 in CNS pathologies. J Neurochem 114:13–27Google Scholar
  3. Butovsky O, Talpalar AE, Ben-Yaakov K, Schwartz M (2005) Activation of microglia by aggregated β-amyloid or lipopolysaccharide impairs MHC-II expression and renders them cytotoxic whereas IFN-γ and IL-4 render them protective. Mol Cell Neurosci 29:381–393CrossRefGoogle Scholar
  4. Chen GY, Nuñez G (2010) Sterile inflammation: sensing and reacting to damage. Nat Rev Immunol 10:826–837CrossRefGoogle Scholar
  5. Colton CA, Wilcock DM (2010) Assessing activation states in microglia. CNS Neurol Disord Drug Targets 9:174–191CrossRefGoogle Scholar
  6. Dementia-statistics (2018) Dementia statistics Alzheimer’s Disease International Accessed 16 May 2017
  7. Dirnagl U, Becker K, Meisel A (2009) Preconditioning and tolerance against cerebral ischaemia: from experimental strategies to clinical use. Lancet Neurol 8:398–412CrossRefGoogle Scholar
  8. Dmowska M, Cybulska R, Schoenborn R, Piersiak T, Jaworska-Adamu J, Gawron A (2010) Behavioural and histological effects of preconditioning with lipopolysaccharide in epileptic rats. Neurochem Res 35:262–272CrossRefGoogle Scholar
  9. Doyle SE, Vaidya SA, O'Connell R, Dadgostar H, Dempsey PW, Wu TT, Rao G, Sun R, Haberland ME, Modlin RL, Cheng G (2002) IRF3 mediates a TLR3/TLR4-specific antiviral gene program. Immunity 17:251–263CrossRefGoogle Scholar
  10. Elliott GT (1998) Monophosphoryl lipid A induces delayed preconditioning against cardiac ischemia-reperfusion injury. J Mol Cell Cardiol 30:3–17CrossRefGoogle Scholar
  11. Frautschy SA, Yang F, Irrizarry M, Hyman B, Saido T, Hsiao K, Cole GM (1998) Microglial response to amyloid plaques in APPsw transgenic mice. Am J Pathol 152:307Google Scholar
  12. Georganopoulou DG, Chang L, Nam J-M, Thaxton CS, Mufson EJ, Klein WL, Mirkin CA (2005) Nanoparticle-based detection in cerebral spinal fluid of a soluble pathogenic biomarker for Alzheimer’s disease. Proc Natl Acad Sci 102:2273–2276CrossRefGoogle Scholar
  13. Grandvaux N, Servant MJ, Sen GC, Balachandran S, Barber GN, Lin R, Hiscott J (2002) Transcriptional profiling of interferon regulatory factor 3 target genes: direct involvement in the regulation of interferon-stimulated genes. J Virol 76:5532–5539CrossRefGoogle Scholar
  14. Grimaldi LME, Zappalà G, Iemolo F, Castellano AE, Ruggieri S, Bruno G, Paolillo A (2014) A pilot study on the use of interferon beta-1a in early Alzheimer’s disease subjects. J Neuroinflammation 11:30CrossRefGoogle Scholar
  15. Gurley C, Nichols J, Liu S, Phulwani NK, Esen N, Kielian T (2008) Microglia and astrocyte activation by toll-like receptor ligands: modulation by PPAR-agonists. PPAR Res 2008:1–15Google Scholar
  16. Heneka MT, Carson MJ, El Khoury J et al (2015) Neuroinflammation in Alzheimer’s disease. Lancet Neurol 14:388–405CrossRefGoogle Scholar
  17. Hesam S, Khoshkholgh-Sima B, Pourbadie HG, Babapour V, Zendedel M, Sayyah M (2018) Monophosphoryl lipid A and Pam3Cys prevent the increase in seizure susceptibility and epileptogenesis in rats undergoing traumatic brain injury. Neurochem Res 43:1978–1985CrossRefGoogle Scholar
  18. Hosseini SM, Pourbadie HG, Sayyah M, Zibaii MI, Naderi N (2018) Neuroprotective effect of monophosphoryl lipid A, a detoxified lipid A derivative, in photothrombotic model of unilateral selective hippocampal ischemia in rat. Behav Brain Res 347:26–36CrossRefGoogle Scholar
  19. Hu F, Ku MC, Markovic D, a Dzaye OD, Lehnardt S, Synowitz M, Wolf SA, Kettenmann H (2014) Glioma-associated microglial MMP9 expression is upregulated by TLR2 signaling and sensitive to minocycline. Int J Cancer 135:2569–2578CrossRefGoogle Scholar
  20. Jacobsen JS, Wu C-C, Redwine JM, Comery TA, Arias R, Bowlby M, Martone R, Morrison JH, Pangalos MN, Reinhart PH, Bloom FE (2006) Early-onset behavioral and synaptic deficits in a mouse model of Alzheimer’s disease. Proc Natl Acad Sci 103:5161–5166CrossRefGoogle Scholar
  21. Janssen B, Vugts DJ, Funke U, Molenaar GT, Kruijer PS, van Berckel BNM, Lammertsma AA, Windhorst AD (2016) Imaging of neuroinflammation in Alzheimer’s disease, multiple sclerosis and stroke: recent developments in positron emission tomography. Biochim Biophys Acta 1862:425–441CrossRefGoogle Scholar
  22. Kamigaki M, Hide I, Yanase Y, Shiraki H, Harada K, Tanaka Y, Seki T, Shirafuji T, Tanaka S, Hide M, Sakai N (2016) The Toll-like receptor 4-activated neuroprotective microglia subpopulation survives via granulocyte macrophage colony-stimulating factor and JAK2/STAT5 signaling. Neurochem Int 93:82–94CrossRefGoogle Scholar
  23. Kochan T, Singla A, Tosi J, Kumar A (2012) Toll-like receptor 2 ligand pretreatment attenuates retinal microglial inflammatory response but enhances phagocytic activity toward Staphylococcus aureus. Infect Immun 80:2076–2088Google Scholar
  24. Ledeboer A, Brevé JJ, Poole S, Tilders FJ, Van Dam AM (2000) Interleukin-10, interleukin-4, and transforming growth factor-β differentially regulate lipopolysaccharide-induced production of pro-inflammatory cytokines and nitric oxide in co-cultures of rat astroglial and microglial cells. Glia 30:134–142CrossRefGoogle Scholar
  25. Lin R, Heylbroeck C, Pitha PM, Hiscott J (1998) Virus-dependent phosphorylation of the IRF-3 transcription factor regulates nuclear translocation, transactivation potential, and proteasome-mediated degradation. Mol Cell Biol 18:2986–2996CrossRefGoogle Scholar
  26. Marsh B, Stevens SL, Packard AE et al (2009) Systemic lipopolysaccharide protects the brain from ischemic injury by reprogramming the response of the brain to stroke: a critical role for IRF3. J Neurosci 29:9839–9849CrossRefGoogle Scholar
  27. Miwa M, Tsuboi M, Noguchi Y, Enokishima A, Nabeshima T, Hiramatsu M (2011) Effects of betaine on lipopolysaccharide-induced memory impairment in mice and the involvement of GABA transporter 2. J Neuroinflammation 8:153Google Scholar
  28. Murpy M, LeVine H III (2010) Alzheimer’s disease and the β-amyloid peptide. J Alzheimers Dis 19:311–323CrossRefGoogle Scholar
  29. Nimmerjahn A, Kirchhoff F, Helmchen F (2005) Resting microglial cells are highly dynamic surveillants of brain parenchyma in vivo. Science 308:1314–1318CrossRefGoogle Scholar
  30. O'neill LA, Golenbock D, Bowie AG (2013) The history of Toll-like receptors—redefining innate immunity. Nat Rev Immunol 13:453–460CrossRefGoogle Scholar
  31. Paranjape GS, Gouwens LK, Osborn DC, Nichols MR (2012) Isolated amyloid-β (1–42) protofibrils, but not isolated fibrils, are robust stimulators of microglia. ACS Chem Neurosci 3:302–311CrossRefGoogle Scholar
  32. Park K, Lee D, Joe E, Kim S, Jin B (2005) Neuroprotective role of microglia expressing interleukin-4. J Neurosci Res 81:397–402CrossRefGoogle Scholar
  33. Pourbadie HG, Sayyah M, Khoshkholgh-Sima B, Choopani S, Nategh M, Motamedi F, Shokrgozar MA (2018) Early minor stimulation of microglial TLR2 and TLR4 receptors attenuates Alzheimer’s disease–related cognitive deficit in rats: behavioral, molecular, and electrophysiological evidence. Neurobiol Aging 70:203–216CrossRefGoogle Scholar
  34. Redwine JM, Kosofsky B, Jacobs RE, Games D, Reilly JF, Morrison JH, Young WG, Bloom FE (2003) Dentate gyrus volume is reduced before onset of plaque formation in PDAPP mice: a magnetic resonance microscopy and stereologic analysis. Proc Natl Acad Sci U S A 100:1381–1386. CrossRefGoogle Scholar
  35. Rego Â, Viana SD, Ribeiro CAF, Rodrigues-Santos P, Pereira FC (2016) Monophosphoryl lipid-A: a promising tool for Alzheimer’s disease toll. J Alzheimers Dis 52:1189–1202CrossRefGoogle Scholar
  36. Reilly JF, Games D, Rydel RE, Freedman S, Schenk D, Young WG, Morrison JH, Bloom FE (2003) Amyloid deposition in the hippocampus and entorhinal cortex: quantitative analysis of a transgenic mouse model. Proc Natl Acad Sci U S A 100:4837–4842. CrossRefGoogle Scholar
  37. Rosenzweig HL, Minami M, Lessov NS, Coste SC, Stevens SL, Henshall DC, Meller R, Simon RP, Stenzel-Poore MP (2007) Endotoxin preconditioning protects against the cytotoxic effects of TNFα after stroke: a novel role for TNFα in LPS-ischemic tolerance. J Cereb Blood Flow Metab 27:1663–1674CrossRefGoogle Scholar
  38. Sisodia SS, Price D (1995) Role of the beta-amyloid protein in Alzheimer’s disease. FASEB J 9:366–370CrossRefGoogle Scholar
  39. Sondag CM, Dhawan G, Combs CK (2009) Beta amyloid oligomers and fibrils stimulate differential activation of primary microglia. J Neuroinflammation 6:1CrossRefGoogle Scholar
  40. Suh H-S, Zhao M-L, Choi N, Belbin TJ, Brosnan CF, Lee SC (2009) TLR3 and TLR4 are innate antiviral immune receptors in human microglia: role of IRF3 in modulating antiviral and inflammatory response in the CNS. Virology 392:246–259CrossRefGoogle Scholar
  41. Tahara K, Kim H-D, Jin J-J, Maxwell JA, Li L, Fukuchi K-i (2006) Role of toll-like receptor signalling in Aβ uptake and clearance. Brain 129:3006–3019CrossRefGoogle Scholar
  42. Tarassishin L, Suh H-S, Lee SC (2011) Interferon regulatory factor 3 plays an anti-inflammatory role in microglia by activating the PI3K/Akt pathway. J Neuroinflammation 8:187CrossRefGoogle Scholar
  43. Thaney VE, O’Neill AM, Hoefer MM, Maung R, Sanchez AB, Kaul M (2017) IFNβ protects neurons from damage in a murine model of HIV-1 associated brain injury. Sci Rep 7:46514CrossRefGoogle Scholar
  44. Toyoda T, Kassell NF, Lee KS (2000) Induction of tolerance against ischemia/reperfusion injury in the rat brain by preconditioning with the endotoxin analog diphosphoryl lipid a. J Neurosurg 92:435–441CrossRefGoogle Scholar
  45. Vartanian KB, Stevens SL, Marsh BJ, Williams-Karnesky R, Lessov NS, Stenzel-Poore MP (2011) LPS preconditioning redirects TLR signaling following stroke: TRIF-IRF3 plays a seminal role in mediating tolerance to ischemic injury. J Neuroinflammation 8:140CrossRefGoogle Scholar
  46. Walsh DM, Klyubin I, Fadeeva JV, Cullen WK, Anwyl R, Wolfe MS, Rowan MJ, Selkoe DJ (2002) Naturally secreted oligomers of amyloid beta protein potently inhibit hippocampal long-term potentiation in vivo. Nature 416:535–539. CrossRefGoogle Scholar
  47. White JA, Manelli AM, Holmberg KH, Van Eldik LJ, LaDu MJ (2005) Differential effects of oligomeric and fibrillar amyloid-β1–42 on astrocyte-mediated inflammation. Neurobiol Dis 18:459–465CrossRefGoogle Scholar
  48. Wood H (2017) Alzheimer disease: twin peaks of microglial activation observed in Alzheimer disease. Nat Rev Neurol 13:129Google Scholar
  49. Yao Y, Li J, Niu Y et al (2015) Resveratrol inhibits oligomeric Aβ-induced microglial activation via NADPH oxidase. Mol Med Rep 12:6133–6139CrossRefGoogle Scholar
  50. Yu JT, Lee CH, Yoo K-Y, Choi JH, Li H, Park OK, Yan B, Hwang IK, Kwon YG, Kim YM, Won MH (2010) Maintenance of anti-inflammatory cytokines and reduction of glial activation in the ischemic hippocampal CA1 region preconditioned with lipopolysaccharide. J Neurol Sci 296:69–78CrossRefGoogle Scholar
  51. Zhao W, Xie W, Xiao Q, Beers DR, Appel SH (2006) Protective effects of an anti-inflammatory cytokine, interleukin-4, on motoneuron toxicity induced by activated microglia. J Neurochem 99:1176–1187CrossRefGoogle Scholar
  52. Zhou X, Spittau B, Krieglstein K (2012) TGFβ signalling plays an important role in IL4-induced alternative activation of microglia. J Neuroinflammation 9:210CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Niloufar Yousefi
    • 1
    • 2
  • Fattah Sotoodehnejadnematalahi
    • 2
  • Nooshin Heshmati-Fakhr
    • 2
  • Mohammad Sayyah
    • 1
  • Masoud Hoseini
    • 3
  • Soheil Ghassemi
    • 1
  • Shayan Aliakbari
    • 1
  • Hamid Gholami Pourbadie
    • 1
    Email author
  1. 1.Department of Physiology and PharmacologyPasteur Institute of IranTehranIran
  2. 2.Department of Biology, School of Basic Science, Science and Research BranchIslamic Azad UniversityTehranIran
  3. 3.Department of Pharmacodynamy and Toxicology, School of PharmacyShahid Beheshti University of Medical SciencesTehranIran

Personalised recommendations