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The flavonoid rutin induces astrocyte and microglia activation and regulates TNF-alpha and NO release in primary glial cell cultures

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Abstract

Astrocyte and microglia cells play an important role in the central nervous system (CNS). They react to various external aggressions by becoming reactive and releasing neurotrophic and/or neurotoxic factors. Rutin is a flavonoid found in many plants and has been shown to have some biological activities, but its direct effects on cells of the CNS have not been well studied. To investigate its potential effects on CNS glial cells, we used both astrocyte primary cultures and astrocyte/microglia mixed primary cell cultures derived from newborn rat cortical brain. The cultures were treated for 24 h with rutin (50 or 100 μmol/L) or vehicle (0.5% dimethyl sulfoxide). Mitochondrial function on glial cells was not evidenced by the MTT test. However, an increased lactate dehydrogenase activity was detected in the culture medium of both culture systems when treated with 100 μmol/L rutin, suggesting loss of cell membrane integrity. Astrocytes exposed to 50 μmol/L rutin became reactive as revealed by glial fibrillary acidic protein (GFAP) overexpression and showed a star-like phenotype revealed by Rosenfeld’s staining. The number of activated microglia expressing OX-42 increased in the presence of rutin. A significant increase of nitric oxide (NO) was observed only in mixed cultures exposed to 100 μmol/L rutin. Enhanced TNFα release was observed in astrocyte primary cultures treated with 100 μmol/L rutin and in mixed primary cultures treated with 50 and 100 μmol/L, suggesting different sensitivity of both activated cell types. These results demonstrated that rutin affects astrocytes and microglial cells in culture and has the capacity to induce NO and TNFα production in these cells. Hence, the impact of these effects on neurons in vitro and in vivo needs to be studied.

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Abbreviations

APC:

astrocyte primary culture

BBB:

blood-brain barrier

CNS:

central nervous system

DMF:

dimethylformamide

ELISA:

enzyme-linked immunosorbent assay

GFAP:

glial fibrillary acidic protein

LDH:

lactate dehydrogenase

MPC:

mixed primary culture

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

PAGE:

polyacrylamide gel electrophoresis

SDS:

sodium dodecyl sulfate

THF:

tetrahydrofuran

TNF:

tumor necrosis factor

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Acknowledgments

This work was supported by grants from Fundação de Amparo à Pesquisa do Estado da Bahia (FAPESB), and Banco do Nordeste do Brasil (BNB). We gratefully acknowledge the research support provided by Programa de Pós-Graduação em Imunologia - UFBA, and Fundação Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).

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Authors and Affiliations

  1. Laboratório de Neuroquímica e Biologia Celular, Departamento de Biofunção, Instituto de Ciências da Saúde, Universidade Federal da Bahia (UFBA), Salvador, Brazil

    A. R. Silva, A. M. Pinheiro, C. S. Souza, S. R. V.-B. Freitas, V. Vasconcellos, R. S. El-Bachá, M. F. D. Costa & S. L. Costa

  2. Laboratório de Imunologia, Departamento de Biointeração, Instituto de Ciências da Saúde, Universidade Federal da Bahia (UFBA), Salvador, BA, 40.110-100, Brazil

    S. M. Freire

  3. Laboratório de Pesquisa em Matéria Médica, Faculdade de Farmácia, UFBA Universidade Federal da Bahia, Salvador, Brazil

    E. S. Velozo

  4. Faculté de Médicine, Val-de-Marne, Université Paris XII, Créteil, France

    M. Tardy

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  1. A. R. Silva
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  2. A. M. Pinheiro
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  3. C. S. Souza
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  4. S. R. V.-B. Freitas
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  5. V. Vasconcellos
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  9. R. S. El-Bachá
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Correspondence to S. L. Costa.

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Silva, A.R., Pinheiro, A.M., Souza, C.S. et al. The flavonoid rutin induces astrocyte and microglia activation and regulates TNF-alpha and NO release in primary glial cell cultures. Cell Biol Toxicol 24, 75–86 (2008). https://doi.org/10.1007/s10565-007-9017-y

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