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Phytochemical study and evaluation of cytotoxicity, mutagenicity, cell cycle kinetics and gene expression of Bauhinia holophylla (Bong.) Steud. in HepG2 cells in vitro

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Abstract

Bauhinia holophylla (Bong.) Steud. (Fabaceae) is a plant used in Brazilian folk medicine to treat diabetes and inflammation. This study evaluated the phytochemical properties, cytotoxic, apoptotic, mutagenic/antimutagenic effects and alterations in gene expression (RNAm) in HepG2 cells treated with the B. holophylla extract. The phytochemical profile highlight the presence of flavonoids isorhamentin and quercetin derivates. The MTT assay was used to evaluate the cytotoxicity of different concentrations for different treatment times. Three concentrations (7.5, 15, 30 µg/mL) were chosen for assessment of apoptosis (AO/EB), mutagenicity (micronucleus), and cell cycle kinetics (flow cytometry). Thereafter, the concentration of 7.5 µg/mL was chosen to evaluate the protective effects against DNA damage induced by benzo[a]pyrene (B[a]P). At concentrations higher than 7.5 µg/mL (between 10 and 50 µg/mL), the extract was cytotoxic, induced apoptosis, and caused antiproliferative effects. However, it did not induce micronucleus and a reduction of apoptotic and micronucleated cells was observed in treatments that included the extract and B[a]P. The protective effect is attributable to the presence of flavonoids, described as antioxidants, inhibitors of DNA adduct and activators of detoxifying enzymes. The results of the present study such as absence of cytotoxic and mutagenic effects and protective effects against known carcinogens suggest that B. holophylla has potential for use soon as herbal medicine.

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Acknowledgements

The authors also wish to thank Dr. E. N. Itano for the use of the spectrophotometer; the Service of Flow Cytometry and Cell Classification of the Institute of Biological Research Clemente Estable (Uruguay) for the use of the FACSVantage Flow Cytometer.

Funding

The authors thank the financial support from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) in the BIOTA Program (2009/52237-9; 2009/16147-5; 2012/01996-0) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior CAPES-PROAP); the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for a scholarship granted to masters student Ribeiro, D.L and for productivity grant to Cólus, I.M.S, Varanda, E.A. and Vilegas, W.

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Authors and Affiliations

  1. Department of General Biology, Biological Sciences Center, State University of Londrina - UEL, Rod Celso Garcia Cid PR 445, Km 380, University Campus, P.O. Box 6001, Londrina, PR, CEP 86057-970, Brazil

    Diego Luis Ribeiro, Heloísa Lizotti Cilião, Ana Flávia Leal Specian, Juliana Mara Serpeloni, Marcelo Tempesta De Oliveira & Ilce Mara Syllos Cólus

  2. Department of Biological Sciences, Faculty of Pharmaceutical Sciences of Araraquara, São Paulo State University - UNESP, Araraquara, SP, Brazil

    Juliana Mara Serpeloni & Eliana Aparecida Varanda

  3. Campus Litoral Paulista, São Paulo State University - UNESP, São Vicente, SP, Brazil

    Wagner Vilegas

  4. Department of Biological Sciences, Faculty of Sciences of Bauru, São Paulo State University – UNESP, Bauru, SP, Brazil

    Luiz Leonardo Saldanha

  5. Department of Botany, Biosciences Institute, São Paulo State University - UNESP, Botucatu, SP, Brazil

    Luiz Leonardo Saldanha & Anne Lígia Dokkedal

  6. Institute of Biological Investigation Clemente Estable, Montevideo, Uruguay

    Wilner Martínez-López

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  1. Diego Luis Ribeiro
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Ribeiro, D.L., Cilião, H.L., Specian, A.F.L. et al. Phytochemical study and evaluation of cytotoxicity, mutagenicity, cell cycle kinetics and gene expression of Bauhinia holophylla (Bong.) Steud. in HepG2 cells in vitro. Cytotechnology 70, 713–728 (2018). https://doi.org/10.1007/s10616-017-0173-5

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