Decrease of the DXR-induced genotoxicity and nongenotoxic effects of Theobroma cacao revealed by micronucleus assay.

M F G Boriollo,V E Alves,G B S Barros,J J Silva,J F Höfling,N M S Oliveira,C T S Dias,T A Silva

doi:10.1590/1519-6984.223687

Abstract

This study evaluated the genotoxicity of lyophilized glycolic extract of Theobroma cacao Linné seeds (TCL), using the micronucleus assay in bone marrow of mice. The interaction between TCL and doxorubicin (DXR) was also analyzed. Experimental groups were evaluated 24-48 h after treatment with N-Nitroso-N-ethylurea (NEU: 50 mg/kg), DXR (5 mg/kg), NaCl (145 mM), TCL (0.5-2 g/kg), and TCL (2 g/kg) in combination with DXR (antigenotoxic assays). Analysis of micronucleated polychromatic erythrocytes (MNPCEs) showed no significant differences between all the treatment doses of TCL and NaCl control. Mice experimentally treated with DXR and NEU significantly induced MNPCEs. However, a significant reduction of MNPCEs was also observed when TCL was administered in combination with the chemotherapeutic agent DXR. The analysis of the PCE/NCE ratio revealed no significant differences between the NaCl control, all doses of TCL, and DXR. However, there were significant differences in the PCE/NCE ratio between positive NEU control and all other treatments. The PCE/NCE ratio observed after treatment with TCL and DXR showed significant differences and intermediate values to controls (NaCl and NEU). This study suggests absence of genotoxicity and cytotoxicity of TCL, regardless of dose, sex, and time. TCL reduced genotoxic effects induced by DXR, suggesting potential antigenotoxic effects.

Highlights

Theobroma cacao L. (Sterculiaceae family) is a species of neotropical origin that grows between a latitude of 18°C North and 15°C South (Murillo et al, 2011)
The micronucleated polychromatic erythrocytes (MNPCEs) (n and %) and the polychromatic erythrocyte (PCE)/ normochromatic erythrocyte (NCE) ratio in the bone marrow of heterogeneous Swiss albinus (Unib: SW) mice were analyzed statistically for each one of the animal groups treated with only T. cacao L. (TCL) – genotoxic assay (0.5, 1.0, 1.5, and 2 g/kg of TCL) – and for the groups treated with phytotherapeutic and chemotherapeutic agent – antigenotoxic assay (2 g/kg of TCL + 5 mg/kg of DXR), as well as for control groups (NaCl 150 mM, NEU 50 mg/kg, or DXR 5 mg/kg)
For animal groups treated with TCL, the MNPCE analysis showed no significant differences (p < 0.05) between all the treatment doses (0.5-2 g/kg) [average values (%) of 0.26 ± 0.10 (24 h) and 0.32 ± 0.34 (48 h) for 500 mg/kg; 0.26 ± 0.12 (24 h) and 0.25 ± 0.14 (48 h) for 1,000 mg/kg; 0.38 ± 0.14 (24 h) and 0.29 ± 0.21 (48 h) for 1,500 mg/kg; and 0.75 ± 0.31 (24 h) and 0.42 ± 0.29 (48 h) for 2,000 mg/kg] and negative control (NaCl) [average values (%) of 0.44 ± 0.10 (24 h) and 0.50 ± 0.08 (48 h)] (Table 1)

Summary

Introduction

Theobroma cacao L. (Sterculiaceae family) is a species of neotropical origin that grows between a latitude of 18°C North and 15°C South (Murillo et al, 2011). (Sterculiaceae family) is a species of neotropical origin that grows between a latitude of 18°C North and 15°C South (Murillo et al, 2011) This arboreal species is a perennial and xenogamic woody plant, presenting high economic impact (ICCO, 2013; Reis et al, 2015) because of its use as raw material in the production of chocolate, cosmetics, fine beverages, jams, ice creams, among others (Souza et al, 2014). The biochemical composition of cocoa powder presents a high content of fibers (26-40%), proteins (15-20%), carbohydrates (15%), lipids (10-24%), as well as minerals and vitamins (Ramiro-Puig and Castell, 2009). About 380 chemical compounds were registered from T. cacao L. (Andújar et al, 2012) and several studies reported its chemical composition and its bioactive compounds are potentially beneficial to human health (Baharum et al, 2014; Ramiro et al, 2007) and to swine livestock production (Jang et al, 2016; Magistrelli et al, 2016), but toxic to dogs depending on the dose (Gwaltney-Brant et al, 2001) and on the CYP1A2 1117C>T genetic polymorphism (Bates et al, 2015)

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