Chemical characterization and cytotoxic, genotoxic, and mutagenic properties of Baccharis trinervis (Lam, Persoon) from Colombia and Brazil
2017
Jaramillo-García, Victoria | Trindade, Cristiano | Lima, Elisiane | Guecheva, Temenouga N. | Villela, Izabel | Martinez-Lopez, Wilner | Corrêa, Dione S. | Ferraz, Alexandre de B.F. | Moura, Sidnei | Sosa, Milton Quintana | Da Silva, Juliana | Henriques, João Antônio Pegas
Baccharis trinervis leaves are used in the traditional medicine for the treatment of high fevers, edema, inflammation, sores and muscle cramps, snakebites and as antiseptics.To investigate the cytotoxic, genotoxic, and mutagenic effects of extracts and fractions of B. trinervis from Brazil and Colombia in Chinese Hamster Ovary (CHO) cells, as well as the mutagenic activity in Salmonella typhimurium.Aqueous extracts (AE) of aerial parts of B. trinervis from Brazil (B) and Colombia (C) were fractioned in ethyl acetate fraction (EAF), butanol extract (BF) and aqueous residue fraction (ARF). Qualitative chemical screening and determination of total flavonoid content were made. Identification of chemical constituents was performed by High Performance Liquid Chromatography (HPLC) and High Resolution Mass Spectrometry (HRMS). For the in vitro tests, CHO cells were treated for 3hours with extracts and fractions of B. trinervis. The cytotoxic activity was evaluated by clonal survival and 3-(4.5-dimethylthiazole-2-yl)−2.5-biphenyl tetrazolium bromide reduction assay (MTT). Also, genotoxic and mutagenic effects were evaluated by the alkaline comet assay and Cytokinesis-blockage micronucleus test (CBMN), respectively. Additionally, Salmonella/microsome assay was carried out to determinate the mutagenic effects in EAF from Brazil and Colombia.Phytochemical analyses indicated the presence of saponins and flavonoids. AE and EAF were the samples with the highest quantity of total flavonoids. HPLC showed the presence of luteolin only in AEC, and caffeic acid, ellagic acid, rosmarinic acid, and rutin were identified in AEB and AEC. However, these compounds had higher concentrations in AEC. The HRMS in positive mode of EAFB and EAFC showed presence of two carboxylic acids, coumarin, and two terpenoids. In addition, in negative mode were identified one terpenoid and two carboxylic acids in AE, BF and ARF of B. trinervis from both countries. Dose-dependent cytotoxic effects were observed in CHO cells treated with B. trinervis extracts and fractions by using clonal survival and MTT at concentrations higher than 0.05mg/mL. All the extracts and fractions induced DNA strand breaks in CHO cells with dose-dependent response, mostly EAFB and EAFC. The EAF from Brazil and Colombia showed mutagenic effect at 0.5mg/mL, while the other fractions did not show a significant difference. No mutagenic effects were found in EAF from both countries by the Salmonella/microsome assay.Phytochemical analyses showed that AE from Colombia presented higher concentrations of the identified compounds than AE from Brazil. Additionally, EAF from both countries showed higher content of the identified compounds than AE, BF an ARF. Furthermore, in vitro toxicological tests showed cytotoxic and genotoxic effects in all extracts and fractions used, although only EAF showed mutagenic effects by CBMN, but not by Salmonella/microsome assay. Our results suggest that flavonoids, phenylpropanoids, coumarins, and diterpenes may be responsible for the cytotoxic, genotoxic and mutagenic effects observed.
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