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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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.
KeywordsAntimutagenic Micronucleus Flavonoids Flow cytometry RT-qPCR
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.
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.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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