Decrease of the DXR-induced genotoxicity and nongenotoxic effects of <i>Theobroma cacao</i> revealed by micronucleus assay

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

doi:10.1590/1519-6984.223687

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Brazilian Journal of Biology

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Original Article • Braz. J. Biol. 81 (2) • Mar-May 2021 • https://doi.org/10.1590/1519-6984.223687 copy

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

Diminuição da genotoxicidade induzida por DXR e efeitos não genotóxicos de Theobroma cacao revelados pelo ensaio de micronúcleo

Authorship SCIMAGO INSTITUTIONS RANKINGS

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.

Keywords:
bone marrow; cacao; cytotoxicity; doxorubicin hydrochloride (DXR); phytotherapeutic; rodents

Treatment	MNPCEs (n)		MNPCEs (%)		PCE / (PCE + NCE)
	24h ^A	48h ^A	24h ^A	48h ^A	24h ^A	48h ^A
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	9 ±2	9 ±1	0.43 ±0.10	0.45 ±0.06	1.00 ±0.01	1.00 ±0.01
♂ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	9 ±3	12 ±1	0.45 ±0.12	0.56 ±0.05	0.99 ±0.01	0.99 ±0.00
Mean ±SD (150 mM NaCl)	9 ±2 ^A	11 ±2 ^A	0.44 ±0.10 ^A	0.50 ±0.08 ^A	0.99 ±0.01 ^A	0.99 ±0.01 ^A
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	28 ±12	34 ±3	1.41 ±0.57	1.66 ±0.07	0.55 ±0.06	0.80 ±0.14
♂ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	65 ±40	36 ±5	3.20 ±1.96	1.83 ±0.27	0.53 ±0.13	0.52 ±0.15
Mean ±SD (NEU 50 mg/kg)	47 ±33 ^B	35 ±4 ^B	2.30 ±1.62 ^B	1.75 ±0.20 ^B	0.54 ±0.09 ^C	0.66 ±0.20 ^C
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	60 ±12	50 ±14	2.86 ±0.57	2.40 ±0.63	0.85 ±0.13	0.97 ±0.02
♂ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	52 ±5	60 ±10	2.49 ±0.26	2.92 ±0.51	0.98 ±0.01	0.97 ±0.02
Mean ±SD (DXR 5 mg/kg)	56 ±9 ^C	55 ±12 ^C	2.68 ±0.44 ^C	2.66 ±0.58 ^C	0.92 ±0.11 ^A	0.97 ±0.02 ^A
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	5 ±1	9 ±10	0.24 ±0.05	0.43 ±0.50	0.99 ±0.01	0.99 ±0.01
♂ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	6 ±3	4 ±1	0.29 ±0.15	0.21 ±0.06	1.00 ±0.01	0.99 ±0.01
Mean ±SD (TCL 500 mg/kg)	6 ±2 ^A	7 ±7 ^A	0.26 ±0.10 ^A	0.32 ±0.34 ^A	1.00 ±0.01 ^A	0.99 ±0.01 ^A
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	3 ±2	7 ±3	0.16 ±0.07	0.33 ±0.15	1.00 ±0.00	1.00 ±0.00
♂ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	8 ±1	3 ±2	0.36 ±0.03	0.16 ±0.07	1.00 ±0.00	0.99 ±0.01
Mean ±SD (TCL 1,000 mg/kg)	6 ±3 ^A	5 ±3 ^A	0.26 ±0.12 ^A	0.25 ±0.14 ^A	1.00 ±0.00 ^A	1.00 ±0.01 ^A
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	8 ±2	9 ±3	0.40 ±0.10	0.45 ±0.14	1.00 ±0.00	1.00 ±0.01
♂ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	7 ±4	3 ±3	0.35 ±0.19	0.13 ±0.12	0.99 ±0.02	0.98 ±0.01
Mean ±SD (TCL 1,500 mg/kg)	8 ±3 ^A	6 ±4 ^A	0.38 ±0.14 ^A	0.29 ±0.21 ^A	1.00 ±0.01 ^A	0.99 ±0.01 ^A
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	16 ±7	11 ±9	0.78 ±0.33	0.54 ±0.40	1.00 ±0.01	1.00 ±0.00
♂ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	15 ±8	6 ±2	0.72 ±0.36	0.29 ±0.08	0.97 ±0.03	0.96 ±0.05
Mean ±SD (TCL 2,000 mg/kg)	16 ±7 ^A	9 ±6 ^A	0.75 ±0.31 ^A	0.42 ±0.29 ^A	0.99 ±0.02 ^A	0.98 ±0.04 ^A
♀ ^A ^(MNPCE); ^A ^{(PCE/NCE ratio)}	20 ±8	9 ±5	0.99 ±0.37	0.42 ±0.24	0.80 ±0.11	0.79 ±0.08
♂ ^A ^(MNPCE); ^A^{(PCE/NCE ratio)}	14 ±6	8 ±7	0.68 ±0.29	0.41 ±0.33	0.85 ±0.18	0.60 ±0.14
Mean ±SD (TCL 2 g/kg + DXR)	17 ±7 ^A	9 ±5 ^A	0.84 ±0.34 ^A	0.42 ±0.26 ^A	0.83 ±0.14 ^B	0.70 ±0.15 ^B

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