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LDH, proliferation curves and cell cycle analysis are the most suitable assays to identify and characterize new phytotherapeutic compounds

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Abstract

Brazilian flora biodiversity has been widely investigated to identify effective and safe phytotherapeutic compounds. Among the investigated plant species, the Byrsonima genus exhibits promising biological activities. This study aimed at evaluating the cytotoxicity of B. correifolia, B. verbascifolia, B. fagifolia and B. intermedia extracts using different assays in two cell lines (primary gastric and HepG2 cells). The different extract concentrations effects on cell viability were assayed using the MTT, aquabluer, neutral red and LDH assays. Non-cytotoxic concentrations were selected to generate cell proliferation curves and to assess cell cycle kinetics by flow cytometry. Byrsonima extracts differentially affected cell viability depending on the metabolic cellular state and the biological parameter evaluated. B. fagifolia and B. intermedia extracts exhibited lower cytotoxic effects than B. correifolia and B. verbascifolia in all assays. The results obtained with LDH and flow cytometry assays were more reliable, suggesting that they can be useful in the screening for herbal medicine and to further characterize these extracts as phytotherapeutic compounds.

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Acknowledgments

This research was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Process No. 2009/52237-9. The authors also thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for the scholarship to Ana Flávia Leal Specian and PROAP; The Conselho Nacional para o Desenvolvimento Científico e Tecnológico (CNPq) for Grants to W. Vilegas, E.A. Varanda and I.M.S. Cólus.

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Authors and Affiliations

  1. Department of General Biology, State University of Londrina, PR 445 Km 380, s/n - Campus Universitário, Londrina, PR, CEP 86057-970, Brazil

    Ana Flávia L. Specian, Juliana M. Serpeloni, Katiuska Tuttis, Diego L. Ribeiro, Heloísa L. Cilião & Ilce M. S. Cólus

  2. Department of Biological Sciences, Faculty of Pharmaceutical Sciences of Araraquara, São Paulo State University “Júlio de Mesquita Filho”, Araraquara, SP, Brazil

    Juliana M. Serpeloni & Eliana A. Varanda

  3. School of Arts, Science and Humanities, São Paulo University, São Paulo, SP, Brazil

    Miriam Sannomiya

  4. Institute of Biosciences, UNESP- Paulista State University, Coastal Campus of São Vicente, São Vicente, SP, Brazil

    Wagner Vilegas

  5. Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay

    Wilner Martinez-Lopez

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  1. Ana Flávia L. Specian
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  2. Juliana M. Serpeloni
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  3. Katiuska Tuttis
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  4. Diego L. Ribeiro
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  5. Heloísa L. Cilião
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  6. Eliana A. Varanda
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  9. Wagner Vilegas
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  10. Ilce M. S. Cólus
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Correspondence to Ana Flávia L. Specian.

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Specian, A.F.L., Serpeloni, J.M., Tuttis, K. et al. LDH, proliferation curves and cell cycle analysis are the most suitable assays to identify and characterize new phytotherapeutic compounds. Cytotechnology 68, 2729–2744 (2016). https://doi.org/10.1007/s10616-016-9998-6

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