Şermin GENÇ, Filiz KURALAY, Başak Bingöl ÇAKIRLI

The Effect of Erythropoietin on Neurotrophic Factors in N9 Murine Microglial Cells

Aim: In this study, we investigated whether interferon gamma (IFNg), lipopolysaccharides (LPS) and amyloid beta (AMYb), as toxic stimulator agents, and erythropoietin (EPO), as a neurotrophic agent, have an effect on the production of the following neurotrophic factors in the N9 murine microglia cell line: neurotrophin 3 (NT3), neurotrophin 4 (NT4), and brain-derived neurotrophic factor (BDNF). Materials and Methods: Microglial cells were incubated with 50 μg/ml AMYb, or 1 μg/ml of LPS plus 100 U/ml recombinant murine IFNg, and/or one of three different concentrations (0.1, 1.0, and 5.0 U/ml) of recombinant mouse EPO for 24 h. Results: EPO 0.1 U/ml dose significantly increased NT4 levels compared to EPO 5.0 U/ml dose (P < 0.05). EPO, in all doses, and AMYb significantly induced NT4 secretion in microglias, while BDNF and NT3 were not changed by AMYb or EPO. LPS + IFNg alone did not change neurotrophic factor levels in any group. However, EPO with LPS and IFNg induced NT4 secretion, especially the 5.0 U/ml dose of EPO. Conclusions: NT4 secretion, which was markedly induced by exposure to both AMYb and EPO in N9 murine microglias, may be an important result for neuronal survival. These results suggest that inflammatory mechanisms in microglia may also involve the neuroprotective response of these cells; this may be a promising area of study of neurodegenerative processes.

Anahtar Kelimeler:

Microglia, N9 cell line, murine, erythropoietin, neurotrophic factors

The Effect of Erythropoietin on Neurotrophic Factors in N9 Murine Microglial Cells

Keywords:

Microglia, N9 cell line, murine, erythropoietin, neurotrophic factors,

PDF

___

Griffin WS. Inflammation and neurodegenerative diseases. Am J
Clin Nutr 2006; 83: 470S-474S. 8.
Genc K, Genc S, Baskın H, Semin I. Erythropoietin decreases
cytotoxicity and nitric oxide formation induced by inflammatory
stimuli in rat oligodendrocytes. Physiol Res 2006; 55: 33-8. 9.
Park MH, Lee SM, Lee JW, Son DJ, Moon DC, Yoon DY et al.
ERK-mediated production of neurotrophic factors by astrocytes
promotes neuronal stem cell differentiation by erythropoietin.
Biochem Biophys Res Commun 2006; 339: 1021-8.
Genc S, Akhisaroglu M, Kuralay F, Genc K. Erythropoietin restores glutathione peroxidase activity in 1-methyl-4-phenyl- 1,2,5,6- tetrahydropyridine-induced neurotoxicity in C-57BL mice and stimulates murine astroglial glutathione peroxidase production in vitro. Neurosci Lett 2002; 321: 73-6.
Genc S, Kuralay F, Genc K, Akhisaroglu M, Fadıloglu S, Yorukoglu K et al. Erythropoietin exerts neuroprotection in 1-methyl-4- phenyl-1,2,3,6–tetrahydropyridine- treated C57BL mice via increasing nitric oxide production. Neurosci Lett 2001; 298: 139- 41.
Siren Al, Fratelli M, Brines M, Geomans C, Casagrande S, Lewczu P et al. Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress. Proc Natl Acad Sci 2001; 98: 4044-9.
Bernaudin M, Bellail A, Marti HH, Yvon A, Vivien D, Duchatelle I et al. Neurons and astrocytes express EPO mRNA: oxygen-sensing mechanisms that involve the redox-state of the brain. Glia 2000; 30: 271-8.
Bernaudin M, Marti HH, Roussel S, Divoux D, Nouvelot A, Mackenzie ET. A potential role for erythropoietin in focal permanent cerebral ischemia in mice. J Cereb Blood Flow Metab 1999; 19: 643-51.
Miwa J, Furukowa S, Nakajima K, Furukowa Y, Kohsaka S. Lipopolysaccharide enhances synthesis of brain-derived neurotrophic factor in cultured rat microglia. J Neurosci Res 1997; 50: 1023-9.
Tanaka J, Koshimura K, Sohmiya M, Murakami Y, Kato Y. Involvement of tetrahydrobiopterin in trophic effect of erythropoietin on PC12 cells. Biochem Biophys Res Commun 2001; 30: 358-62.
Caldwell MA, He X, Wilkie N, Pollack S, Marshall G, Wafford KA et al. Growth factors regulate the survival and fate of cells derived from human neurospheres. Nat Biotechnol 2001; 19: 475-9.
Darlington CL. Astrocytes as targets for neuroprotective drugs. Curr Opin Investig Drugs 2005; 6: 700.
Lahteinen S, Pitkanen A, Knuuttila J, Toronen P, Castren E. Brain- derived neurotrophic factor signaling modifies hippocampal gene expression during epileptogenesis in transgenic mice. Eur J Neurosci 2004; 19: 3245-54.
Burbach GJ, Hellweg R, Haas CA, Del Turco D, Deicke U, Abramowski D et al. Induction of brain-derived neurotrophic factor in plaque-associated glial cells of aged APP23 transgenic mice. J Neurosci 2004; 24: 2421-30.
Duman RS. Role of neurotrophic factors in the etiology and treatment of mood disorders. Neuromolecular Med 2004; 5: 11- 25.
Fraher J. Axons and glial interfaces: ultrastructural studies. Anat 2002; 200: 415-30.
Chan JR, Cosgaya JM, Wu YJ, Shooter EM. Neurotrophins are key mediators of the myelination program in the peripheral nervous system. Proc Natl Acad Sci U S A 2001; 98: 14661-8.
Dougherty KD, Dreyfus CF, Black IB. Brain-derived neurotrophic factor in astrocytes, oligodendrocytes, and microglia/ macrophages after spinal cord injury. Neurobiol Dis 2000; 7: 574-85.
Teismann P, Tieu K, Cohen O, Choi DK, Wu DC, Marks D et al. Pathogenic role of glial cells in Parkinson’s disease. Mov Disord 2003; 18: 121-9. Zhang J, Geula C, Lu C, Koziel H, Hatcher LM, Roien FJ. Neurotrophins regulate proliferation and survival of two microglial cell lines in vitro. Exp Neurol 2003; 183: 469-81.
Toyomoto M, Ohta M, Okumura K, Yano H, Matsumoto K, Inoue S et al. Prostaglandins are powerful inducers of NGF and BDNF production in mouse astrocyte cultures. FEBS Lett 2004; 562: 211-5.
Mallat M, Houlgatte R, Brachet P, Prochiantz A. Lipopolysaccharide-stimulated rat brain macrophages release NGF in vitro. Dev Biol 1989; 133: 309-11.
Heese K, Hock C, Otten U. Inflammatory signals induce neurotrophin expression in human microglial cells. J Neurochem 1998; 70: 699-707.
Frade JM, Barde YA. Nerve growth factor: two receptors, multiple functions. Bioessays 1998; 20: 137-45.
Miwa T, Furukawa S, Nakajima K, Furukawa Y, Kohsaka S. Lipopolysaccharide enhances synthesis of brain-derived neurotrophic factor in cultured rat microglia. J Neurosci Res 1997; 50: 1023-9.
Elkabes S, Peng L, Black IB. Lipopolysaccharide differentially regulates microglial trk receptor and neurotrophin expression. J Neurosci Res 1998; 54: 117-22.
Wang L, Zhang Z, Wang Y, Zhang R, Chopp M. Treatment of stroke with erythropoietin enhances neurogenesis and angiogenesis and improves neurological function in rats. Stroke 2004; 35: 1732-7.
Zhang J, Li Y, Cui Y, Chen J, Lu M, Elias SB et al. Erythropoietin treatment improves neurological functional recovery in EAE mice. Brain Res 2005; 1034: 34-9.
Viviani B, Bartesaghi S, Corsini E, Villa P, Ghezzi P, Garau A et al. Erythropoietin protects primary hippocampal neurons increasing the expression of brain-derived neurotrophic factor. J Neurochem 2005; 93: 412-21.
Olivieri G, Otten U, Meier F, Baysang G, Dimitriades-Schmutz B, Müller-Spahn F et al. Beta-amyloid modulates tyrosine kinase B receptor expression in SHSY5Y neuroblastoma cells: influence of the antioxidant melatonin. Neurosci 2003; 120: 659-65.
Mizuno T, Kuno R, Nitta A, Nabeshima T, Zhang G, Kawanokuchi J et al. Protective effects of nicergoline against neuronal cell death induced by activated microglia and astrocytes. Brain Res 2005; 1066: 78-85.
Wen TC, Sadamoto Y, Tanaka J, Zhu PX, Nakata K, Ma YJ et al. Erythropoietin protects neurons against chemical hypoxia and cerebral ischemic injury by up-regulating Bcl-xl expression. J Neurosci Res 2002; 67: 795-803.
Chong ZZ, Li F, Maiese K. Erythropoietin requires NF-kappaB and its nuclear translocation to prevent early and late apoptotic neuronal injury during beta-amyloid toxicity. Curr Neurovasc Res 2005; 2: 387-99.
Chong ZZ, Kang JQ, Maiese K. Erythropoietin is a novel vascular protectant through activation of Akt1 and mitochondrial modulation of cysteine proteases. Circulation 2002; 106: 2973–9.
Chong ZZ, Kang JQ, Maiese K. Erythropoietin fosters both intrinsic and extrinsic neuronal protection through modulation of microglia, Akt1, Bad, and caspase-mediated pathways. Br J Pharmacol 2003; 138: 1107–18.
Kawakami M, Sekiguchi M, Sato K, Kozaki S, Takahashi M. Erythropoietin receptor-mediated inhibition of exocytotic glutamate release confers neuroprotection during chemical ischemia. J Biol Chem 2001; 276: 39469–75.