Reduction of doxorubicin-induced genotoxicity by Handroanthus impetiginosus in mouse bone marrow revealed by micronucleus assay

Abstract

Handroanthus impetiginosus has long been used in traditional medicine and various studies have determined the presence of bioactive chemical compounds and potential phytotherapeutics. In this study, the genotoxicity of the lyophilized tincture of H. impetiginosus bark (THI) was evaluated in mouse bone marrow using micronucleus assays. The interaction between THI and genotoxic effects induced by the chemotherapeutic agent, doxorubicin (DXR), was also analyzed. Experimental groups were evaluated 24 to 48 h after treatment with N-nitroso-N-ethylurea (NEU; 50 mg/kg), DXR (5 mg/kg), sodium chloride (NaCl; 150 mM), and THI (0.5-2 g/kg). Antigenotoxic assays were carried out using THI (0.5 g/kg) in combination with NEU or DXR. Analysis of the micronucleated polychromatic erythrocytes (MNPCEs) indicated no significant differences between treatment doses of THI (0.5-2 g/kg) and NaCl. Polychromatic erythrocyte (PCE) to normochromatic erythrocyte (NCE) ratios did not indicate any statistical differences between DXR and THI or NaCl, but there were differences between THI and NaCl. A significant reduction in MNPCEs and PCE/NCE ratios was observed when THI was administered in combination with DXR. This study suggested the absence of THI genotoxicity that was dose-, time-, and gender-independent and the presence of moderate systemic toxicity that was dose-independent, but time-and gender-dependent. The combination of THI and DXR also suggested antigenotoxic effects, indicating that THI reduced genotoxic effects induced by chemotherapeutic agents.Keywords: Handroanthus impetiginosus (Mart. ex DC.) Mattos, phytotherapics, doxorubicin (DXR), micronucleus assay, bone marrow. Redução da genotoxicidade induzida pela doxorrubicina por Handroanthus impetiginosus na medula óssea de camundongos revelada pelo ensaio do micronúcleo ResumoHandroanthus impetiginosus tem sido usada durante um longo período pela medicina tradicional e vários estudos têm demonstrados a presença de compostos químicos e potencial fitoterapêutico. Esta pesquisa avaliou a genotoxicidade da tintura da casca liofilizada de H. impetiginosus (THI) usando o ensaio do micronúcleo em medula óssea de camundongos. A interação entre THI e os efeitos genotóxicos induzidos pelo quimioterápico doxorrubicina (DXR) também foram analisados. Grupos experimentais foram analisados a 24-48 h após o tratamento com N-Nitroso-N-etiluréia (NEU; 50 mg/kg), DXR (5 mg/kg), NaCl (150 mM) e THI (0,5-2 g/kg). O ensaio antigenotóxico foi conduzido utilizando THI (0,5 g/kg) em combinação com NEU ou DXR. A análise de eritrócitos policromáticos micronucleados (EPCMNs) não mostrou diferenças significativas entre as doses de tratamento (0,5-2 g/kg) e NaCl. As proporções de eritrócitos policromáticos (EPC)/eritrócitos normocromáticos (ENC) não revelaram diferenças estatísticas entre DXR e THI ou NaCl, porém houve diferenças entre THI e NaCl. Uma redução significativa em EPCMNs e na razão EPC/ENC foi observada quando THI foi administrado em combinação com DXR. Essa pesquisa sugere ausência de ...

Antihelminthic action of the Anethum graveolens essential oil on Haemonchus contortus eggs and larvae

“…For a product to be considered a good infection control agent, causing no harm to the host is as important as the deleterious effect of phytotherapic on the parasite. Although the toxic effect of some plants that have already been described, other studies have already shown an opposite effect, a protection against the toxicity of other chemicals (Boriollo et al, 2018). In a study with A. graveolens leaf extract containing silver nanoparticles, Kalangi et al (2016) performed the same cell viability assay and observed at the highest concentration of 100 mg a viability of 82%.…”

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

“…Healthy, heterogeneous, young adult male and female Swiss albinus (Unib: SW) mice (between 7 and 12 weeks -pubescent period), with a body weight between 30 g and 40 g (i.e., the variation weight between the animals, for each sex, should not exceed the ± 20% of mean mass) were provided by CEMIB (Multidisciplinary Center for Biological Research -UNICAMP), and erythrocytes from the bone marrow of these mice were used in the micronucleus assay (Boriollo et al, 2014b(Boriollo et al, , 2018OECD, 2016;Venkatesh et al, 2007). The animals were kept in groups of the same sex, in polypropylene boxes, in an air-conditioned environment to 22°C ± 3°C, with relative air humidity of 50-60% ± 10%, and with 12-hour day-night cycles (i.e., 12 h light and 12 h dark).…”

“…Groups of animals (consisting of 5 males and 5 females each) were treated using a single dosing regimen administered by gavage (phytotherapeutic and negative control) or intraperitoneally (chemotherapeutic and positive control) and two euthanasia times (24 and 48 h), based on a regulatory recommendation regarding the in vivo micronucleus assay (Boriollo et al, 2014b(Boriollo et al, , 2018OECD, 2016;Venkatesh et al, 2007): ▪ Control groups: 150 mM NaCl (negative control), 50 mg/kg of N-Nitroso-N-ethylurea (positive control: NEU, Sigma N8509, CAS no. 759-73-9), and 5 mg/kg of doxorubicin hydrochloride (chemotherapeutic: DXR, Eurofarma Laboratórios Ltda., CAS no.…”

“…As far as genotoxicity studies are concerned, the in vivo micronucleus (MN) assay in rodent bone marrow plays a crucial role in the test battery aimed at identifying hazardous mutagens (Mateuca et al, 2006); this assay is especially suited to assessing mutagenic hazards because it allows consideration of multiple factors, such as in vivo metabolism, pharmacokinetics, and DNA repair processes, even though these processes vary among species, tissues, and genetic endpoints (OECD, 2016). In addition, understanding the genotoxic effects induced by phytotherapeutics and foods employing the mammalian in vivo MN assay has been the goal of several research groups (Venkatesh et al, 2007;Alves et al, 2013;Boriollo et al, 2014bBoriollo et al, , 2018.…”

“…, and evaporation bottle (RV 10.85 Evaporation Flask, NS 29/32-2L, IKA  Works, Inc., USA). The final product was transferred to 1 L reaction bottle (SCHOTT  DURAN  ) and kept at −20°C for 24 hours, to evaluate the freezing of the final product and the efficacy of the solvent evaporation process (Boriollo et al, 2014b(Boriollo et al, , 2018.…”