Lipopolysaccharide modulates astrocytic S100B secretion: a study in cerebrospinal fluid and astrocyte cultures from rats

Journal of Neuroinflammation

Carlos-Alberto Gonçalves¹  Marina C Leite¹  Letícia Rodrigues¹  Douglas S Engelke¹  Elisa Negri¹  Carollina Da Ré¹  Fabiana Galland¹  Lucas S Tortorelli¹  Maria Cristina Guerra¹ 

[1] Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Ramiro Barcelos, 2600- Anexo, 90035-003, Porto Alegre, Brazil

关键词: S100B;    TLR4;    LPS;    glutathione;    GFAP;    astrocyte;   

Others  :  1213094
DOI  :  10.1186/1742-2094-8-128

 received in 2010-12-02, accepted in 2011-10-04,  发布年份 2011

【 摘 要 】

Background

Inflammatory responses in brain are primarily mediated by microglia, but growing evidence suggests a crucial importance of astrocytes. S100B, a calcium-binding protein secreted by astrocytes, has properties of a neurotrophic or an inflammatory cytokine. However, it is not known whether primary signals occurring during induction of an inflammatory response (e.g. lipopolysaccharide, LPS) directly modulate S100B.

Methods

In this work, we evaluated whether S100B levels in cerebrospinal fluid (CSF) and serum of Wistar rats are affected by LPS administered by intraperitoneal (IP) or intracerebroventricular (ICV) injection, as well as whether primary astrocyte cultures respond directly to lipopolysaccharide.

Results

Our data suggest that S100B secretion in brain tissue is stimulated rapidly and persistently (for at least 24 h) by ICV LPS administration. This increase in CSF S100B was transient when LPS was IP administered. In contrast to these S100B results, we observed an increase in in TNFα levels in serum, but not in CSF, after IP administration of LPS. In isolated astrocytes and in acute hippocampal slices, we observed a direct stimulation of S100B secretion by LPS at a concentration of 10 μg/mL. An involvement of TLR4 was confirmed by use of specific inhibitors. However, lower levels of LPS in astrocyte cultures were able to induce a decrease in S100B secretion after 24 h, without significant change in intracellular content of S100B. In addition, after 24 h exposure to LPS, we observed a decrease in astrocytic glutathione and an increase in astrocytic glial fibrillary acidic protein.

Conclusions

Together, these data contribute to the understanding of the effects of LPS on astrocytes, particularly on S100B secretion, and help us to interpret cerebrospinal fluid and serum changes for this protein in neuroinflammatory diseases. Moreover, non-brain S100B-expressing tissues may be differentially regulated, since LPS administration did not lead to increased serum levels of S100B.

【 授权许可】

   
2011 Guerra et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Donato R, Sorci G, Riuzzi F, Arcuri C, Bianchi R, Brozzi F, Tubaro C, Giambanco I: S100B's double life: intracellular regulator and extracellular signal. Biochim Biophys Acta 2009, 1793:1008-1022.
• [2]Pinto SS, Gottfried C, Mendez A, Goncalves D, Karl J, Goncalves CA, Wofchuk S, Rodnight R: Immunocontent and secretion of S100B in astrocyte cultures from different brain regions in relation to morphology. FEBS Lett 2000, 486:203-207.
• [3]Tramontina AC, Tramontina F, Bobermin LD, Zanotto C, Souza DF, Leite MC, Nardin P, Gottfried C, Goncalves CA: Secretion of S100B, an astrocyte-derived neurotrophic protein, is stimulated by fluoxetine via a mechanism independent of serotonin. Prog Neuropsychopharmacol Biol Psychiatry 2008, 32:1580-1583.
• [4]Sakatani S, Seto-Ohshima A, Shinohara Y, Yamamoto Y, Yamamoto H, Itohara S, Hirase H: Neural-activity-dependent release of S100B from astrocytes enhances kainate-induced gamma oscillations in vivo. J Neurosci 2008, 28:10928-10936.
• [5]Buyukuysal RL: Protein S100B release from rat brain slices during and after ischemia: comparison with lactate dehydrogenase leakage. Neurochem Int 2005, 47:580-588.
• [6]Nardin P, Tramontina F, Leite MC, Tramontina AC, Quincozes-Santos A, de Almeida LM, Battastini AM, Gottfried C, Goncalves CA: S100B content and secretion decrease in astrocytes cultured in high-glucose medium. Neurochem Int 2007, 50:774-782.
• [7]Leite MC, Brolese G, de Almeida LM, Pinero CC, Gottfried C, Goncalves CA: Ammonia-induced alteration in S100B secretion in astrocytes is not reverted by creatine addition. Brain Res Bull 2006, 70:179-185.
• [8]Steiner J, Bernstein HG, Bielau H, Berndt A, Brisch R, Mawrin C, Keilhoff G, Bogerts B: Evidence for a wide extra-astrocytic distribution of S100B in human brain. BMC Neurosci 2007, 8:2. BioMed Central Full Text
• [9]Haimoto H, Kato K, Suzuki F, Nagura H: The ultrastructural changes of S-100 protein localization during lipolysis in adipocytes. An immunoelectron-microscopic study. Am J Pathol 1985, 121:185-191.
• [10]Rothermundt M, Peters M, Prehn JH, Arolt V: S100B in brain damage and neurodegeneration. Microsc Res Tech 2003, 60:614-632.
• [11]Sen J, Belli A: S100B in neuropathologic states: the CRP of the brain? J Neurosci Res 2007, 85:1373-1380.
• [12]de Souza DF, Leite MC, Quincozes-Santos A, Nardin P, Tortorelli LS, Rigo MM, Gottfried C, Leal RB, Goncalves CA: S100B secretion is stimulated by IL-1beta in glial cultures and hippocampal slices of rats: Likely involvement of MAPK pathway. J Neuroimmunol 2009, 206:52-57.
• [13]Kim SH, Smith CJ, Van Eldik LJ: Importance of MAPK pathways for microglial pro-inflammatory cytokine IL-1 beta production. Neurobiol Aging 2004, 25:431-439.
• [14]Bianchi R, Giambanco I, Donato R: S100B/RAGE-dependent activation of microglia via NF-kappaB and AP-1 Co-regulation of COX-2 expression by S100B, IL-1beta and TNF-alpha. Neurobiol Aging 2010, 31:665-677.
• [15]Ponath G, Schettler C, Kaestner F, Voigt B, Wentker D, Arolt V, Rothermundt M: Autocrine S100B effects on astrocytes are mediated via RAGE. J Neuroimmunol 2007, 184:214-222.
• [16]Belanger M, Magistretti PJ: The role of astroglia in neuroprotection. Dialogues Clin Neurosci 2009, 11:281-295.
• [17]Eng LF, Ghirnikar RS, Lee YL: Glial fibrillary acidic protein: GFAP-thirty-one years (1969-2000). Neurochem Res 2000, 25:1439-1451.
• [18]Hauss-Wegrzyniak B, Lukovic L, Bigaud M, Stoeckel ME: Brain inflammatory response induced by intracerebroventricular infusion of lipopolysaccharide: an immunohistochemical study. Brain Res 1998, 794:211-224.
• [19]Farina C, Aloisi F, Meinl E: Astrocytes are active players in cerebral innate immunity. Trends Immunol 2007, 28:138-145.
• [20]Carpentier PA, Duncan DS, Miller SD: Glial toll-like receptor signaling in central nervous system infection and autoimmunity. Brain Behav Immun 2008, 22:140-147.
• [21]Liao CK, Wang SM, Chen YL, Wang HS, Wu JC: Lipopolysaccharide-induced inhibition of connexin43 gap junction communication in astrocytes is mediated by downregulation of caveolin-3. Int J Biochem Cell Biol 2010, 42:762-770.
• [22]Vergara D, Martignago R, Bonsegna S, De Nuccio F, Santino A, Nicolardi G, Maffia M: IFN-beta reverses the lipopolysaccharide-induced proteome modifications in treated astrocytes. J Neuroimmunol 2010, 221:115-120.
• [23]Brahmachari S, Fung YK, Pahan K: Induction of glial fibrillary acidic protein expression in astrocytes by nitric oxide. J Neurosci 2006, 26:4930-4939.
• [24]Leite MC, Galland F, de Souza DF, Guerra MC, Bobermin L, Biasibetti R, Gottfried C, Goncalves CA: Gap junction inhibitors modulate S100B secretion in astrocyte cultures and acute hippocampal slices. J Neurosci Res 2009.
• [25]Rodrigues L, Biasibetti R, Swarowsky A, Leite MC, Quincozes-Santos A, Quilfeldt JA, Achaval M, Goncalves CA: Hippocampal alterations in rats submitted to streptozotocin-induced dementia model are prevented by aminoguanidine. J Alzheimers Dis 2009, 17:193-202.
• [26]Gottfried C, Valentim L, Salbego C, Karl J, Wofchuk ST, Rodnight R: Regulation of protein phosphorylation in astrocyte cultures by external calcium ions: specific effects on the phosphorylation of glial fibrillary acidic protein (GFAP), vimentin and heat shock protein 27 (HSP27). Brain Res 1999, 833:142-149.
• [27]Nardin P, Tortorelli L, Quincozes-Santos A, de Almeida LM, Leite MC, Thomazi AP, Gottfried C, Wofchuk ST, Donato R, Goncalves CA: S100B Secretion in Acute Brain Slices: Modulation by Extracellular Levels of Ca(2+) and K (+). Neurochem Res 2009.
• [28]Leite MC, Galland F, Brolese G, Guerra MC, Bortolotto JW, Freitas R, Almeida LM, Gottfried C, Goncalves CA: A simple, sensitive and widely applicable ELISA for S100B: Methodological features of the measurement of this glial protein. J Neurosci Methods 2008, 169:93-99.
• [29]Tramontina F, Leite MC, Cereser K, de Souza DF, Tramontina AC, Nardin P, Andreazza AC, Gottfried C, Kapczinski F, Goncalves CA: Immunoassay for glial fibrillary acidic protein: antigen recognition is affected by its phosphorylation state. J Neurosci Methods 2007, 162:282-286.
• [30]Browne RW, Armstrong D: Reduced glutathione and glutathione disulfide. Methods Mol Biol 1998, 108:347-352.
• [31]Goncalves CA, Concli Leite M, Nardin P: Biological and methodological features of the measurement of S100B, a putative marker of brain injury. Clin Biochem 2008, 41:755-763.
• [32]Garnier Y, Berger R, Alm S, von Duering MU, Coumans AB, Michetti F, Bruschettini M, Lituania M, Hasaart TH, Gazzolo D: Systemic endotoxin administration results in increased S100B protein blood levels and periventricular brain white matter injury in the preterm fetal sheep. Eur J Obstet Gynecol Reprod Biol 2006, 124:15-22.
• [33]Rosengarten B, Wolff S, Klatt S, Schermuly RT: Effects of inducible nitric oxide synthase inhibition or norepinephrine on the neurovascular coupling in an endotoxic rat shock model. Crit Care 2009, 13:R139. BioMed Central Full Text
• [34]van den Boogaard M, Ramakers BP, van Alfen N, van der Werf SP, Fick WF, Hoedemaekers CW, Verbeek MM, Schoonhoven L, van der Hoeven JG, Pickkers P: Endotoxemia-induced inflammation and the effect on the human brain. Crit Care 2010, 14:R81. BioMed Central Full Text
• [35]Gorina R, Font-Nieves M, Marquez-Kisinousky L, Santalucia T, Planas AM: Astrocyte TLR4 activation induces a proinflammatory environment through the interplay between MyD88-dependent NFkappaB signaling, MAPK, and Jak1/Stat1 pathways. Glia 2011, 59:242-255.
• [36]Banks WA, Robinson SM: Minimal penetration of lipopolysaccharide across the murine blood-brain barrier. Brain Behav Immun 2010, 24:102-109.
• [37]Edwards MM, Robinson SR: TNF alpha affects the expression of GFAP and S100B: implications for Alzheimer's disease. J Neural Transm 2006, 113:1709-1715.
• [38]Leite M, Frizzo JK, Nardin P, de Almeida LM, Tramontina F, Gottfried C, Goncalves CA: Beta-hydroxy-butyrate alters the extracellular content of S100B in astrocyte cultures. Brain Res Bull 2004, 64:139-143.
• [39]Sheng JG, Ito K, Skinner RD, Mrak RE, Rovnaghi CR, Van Eldik LJ, Griffin WS: In vivo and in vitro evidence supporting a role for the inflammatory cytokine interleukin-1 as a driving force in Alzheimer pathogenesis. Neurobiol Aging 1996, 17:761-766.
• [40]Hinkle DA, Harney JP, Cai A, Hilt DC, Yarowsky PJ, Wise PM: Basic fibroblast growth factor-2 and interleukin-1 beta regulate S100 beta expression in cultured astrocytes. Neuroscience 1998, 82:33-41.
• [41]Buffo A, Rite I, Tripathi P, Lepier A, Colak D, Horn AP, Mori T, Gotz M: Origin and progeny of reactive gliosis: A source of multipotent cells in the injured brain. Proc Natl Acad Sci USA 2008, 105:3581-3586.
• [42]Dringen R, Hirrlinger J: Glutathione pathways in the brain. Biol Chem 2003, 384:505-516.
• [43]Liu L, Li Y, Van Eldik LJ, Griffin WS, Barger SW: S100B-induced microglial and neuronal IL-1 expression is mediated by cell type-specific transcription factors. J Neurochem 2005, 92:546-553.
• [44]Bianchi ME: DAMPs, PAMPs and alarmins: all we need to know about danger. J Leukoc Biol 2007, 81:1-5.
• [45]Peri F, Piazza M, Calabrese V, Damore G, Cighetti R: Exploring the LPS/TLR4 signal pathway with small molecules. Biochem Soc Trans 2010, 38:1390-1395.
• [46]Goncalves CA, Leite MC, Guerra MC: Adipocytes as an Important Source of Serum S100B and Possible Roles of This Protein in Adipose Tissue. Cardiovasc Psychiatry Neurol 2010, 2010:790431.
• [47]Cirillo C, Sarnelli G, Esposito G, Grosso M, Petruzzelli R, Izzo P, Cali G, D'Armiento FP, Rocco A, Nardone G, et al.: Increased mucosal nitric oxide production in ulcerative colitis is mediated in part by the enteroglial-derived S100B protein. Neurogastroenterol Motil 2009, 21:1209-e1112.
• [48]Unden J, Christensson B, Bellner J, Alling C, Romner B: Serum S100B levels in patients with cerebral and extracerebral infectious disease. Scand J Infect Dis 2004, 36:10-13.