This study investigated the chemical composition, immunomodulatory and antioxidant activities, and histopathological analysis of mice tissues treated with methanolic extract from R. marina poison. Marinobufagin, telecinobufagin and bufalin were identified in the chemical profile. The biochemical results demonstrated an effect between doses in the period of 7 days, an immunomodulatory effect was observed regarding the production capacity of interleukin (IL)-12p70 and tumor necrosis factor (TNF)-α at 7 and 30 days, respectively. The lower dose suggests better bioactivity to the treated animal than the higher dose. Histopathological analyses of the lung, heart, kidney and liver showed tissue damage in all organs, mainly in the lung, and were proportional to the dose and the treatment period. We observed that the treatment modulated cytokine production, and therefore this effect may be related to the tissue damage observed. This study demonstrates a positive effect in the antioxidant and immune system, indicating that the molecules found in the extract have biotechnological potential.

RESUMO A secreção cutânea de anuros da família Bufonidae tem grande potencial na busca de novos compostos ativos para utilização como fármacos candidatos no tratamento de algumas doenças, entre elas o câncer. Neste contexto, este estudo teve como objetivo avaliar a atividade citotóxica e antimutagênica dos extratos da secreção da glândula parótida de Rhinella marina e Rhaebo guttatus, bem como a análise bioquímica de transaminases e creatinina séricas, para avaliar dano hepático e renal, respectivamente. A avaliação de citotoxicidade foi realizada pelo método colorimétrico baseado no MTT (3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide), com diferentes concentrações dos extratos em culturas de células do Tumor de Walker ou células esplênicas de rato e camundongo. O teste do micronúcleo foi realizado com camundongos Swiss machos que receberam tratamento oral com os extratos durante 15 dias, seguido de tratamento intraperitoneal com N-etil-N-nitrosuréia (50 mg kg-1). A frequência de eritrócitos policromáticos micronucleados (PCEMN) foi determinada em medula óssea. Os extratos apresentaram ação citotóxica nas células avaliadas. Houve uma redução significativa na frequência de PCEMN (R. marina = 56% e R. guttatus = 75%, p < 0,001), observando-se um potencial antimutagênico dos extratos. Os grupos tratados somente com os extratos apresentaram um aumento na frequência de PCEMNs, evidenciando um potencial mutagênico. As análises bioquímicas não apresentaram diferença significativa entre os tratamentos. Assim, nas condições experimentais testadas, as secreções cutâneas de R. marina e R. guttatus apresentaram potencial quimiopreventivo para câncer.

ABSTRACT The skin secretion from toads of the Bufonidae family has great potential in the search for new active compounds to be used as drug candidates in treating some diseases, among them cancer. In this context, this study aimed to evaluate the cytotoxic and antimutagenic activity of the parotoid gland secretion extracts of Rhinella marina and Rhaebo guttatus, as well as biochemically analyze transaminases and serum creatinine for liver and renal damage, respectively. Cytotoxicity was performed by the colorimetric method based on MTT (3- [4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide) with different concentrations of the extracts in Walker or splenic tumor cell cultures from rats and mice. The micronucleus test was performed with male Swiss mice treated orally with the extracts for 15 days, and then intraperitoneally with N-ethyl-N-nitrosurea (50 mg kg-1). Micronucleated polychromatic erythrocytes (MNPCE) were evaluated in bone marrow. The extracts showed cytotoxic activity in the evaluated cells. There was a significant reduction in the frequency of MNPCE (R. marina = 56% and R. guttatus = 75%, p < 0.001), indicating antimutagenic potential of the extracts. The groups treated only with extract showed an increase in MNPCE frequency, evidencing mutagenic potential. Biochemical analyzes showed no significant difference between treatments. Thus, under our experimental conditions, the extracts of R. marina and R. guttatus skin secretions presented chemopreventive potential for cancer.

RESUMO Copaifera multijuga, ou copaíba, é utilizada popularmente como chá para o tratamento de diversas afecções, o que se deve à presença de substâncias antioxidantes em sua composição, que protegem as células contra danos causados pelos radicais livres. O óleo-resina da árvore é usado como antiinflamatório e antitumoral. O presente estudo avaliou o efeito antioxidante do extrato etanólico da casca da copaíba sobre camundongos Swiss machos inoculados com tumor sólido de Ehrlich. Os camundongos foram inoculados subcutaneamente com 1x106 células de tumor de Ehrlich e foram tratados, via gavagem durante 30 dias, com doses de extrato etanólico de copaíba variando de 100 a 200 mg kg-1. Realizou-se análise bioquímica dos antioxidantes enzimáticos [superóxido dismutase (SOD), catalase (CAT), glutationa-S-transferase (GST)], antioxidantes não-enzimáticos [glutationa reduzida (GSH) e ácido ascórbico (ASA)], substâncias reativas ao ácido tiobarbitúrico (TBARS) e carbonilação proteica (carbonil) em diferentes tecidos e apresentando resultados significativos. O extrato administrado na concentração de 200 mg kg-1 apresentou melhor capacidade antioxidante no fígado, aumentando a CAT, GST, GSH e diminuindo TBARS, além de aumentar a atividade da CAT e da carbonilação proteica no tecido cerebral. Os resultados mostram que o extrato de copaíba foi capaz de reverter o estresse oxidativo causado pelo tumor sólido de Ehrlich, provavelmente por conter compostos antioxidantes, e possivelmente teve um efeito antineoplásico após 30 dias de tratamento.

ABSTRACT Copaifera multijuga, commonly known as copaiba, is popularly used in the form of tea for various conditions due to the presence of antioxidant substances in its composition, which protect cells against damage caused by free radicals. Its oleoresin is also used as an anti-inflammatory and antitumoral agent. The present study investigated the antioxidant effect of the ethanolic extract of copaiba stem bark on Swiss mice inoculated with solid Ehrlich tumors. Mice were inoculated subcutaneously with 1x106 Ehrlich’s tumor cells and treated via gavage with ethanolic extract of copaiba for thirty days, with doses varying between 100 and 200 mg kg-1. Biochemical analyses of enzymatic antioxidants [superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST)], non-enzymatic antioxidants [reduced glutathione (GSH) and ascorbic acid (ASA)], substances reactive to thiobarbituric acid (TBARS) and protein carbonylation (carbonyl) in different tissues were significantly affected. The extract administered at 200 mg kg-1 presented higher antioxidant capacity in the liver, increased CAT, GST, GSH and decreased TBARS, as well as increased CAT activity and protein carbonylation in brain tissue. The results showed that the copaiba extract was able to reverse the oxidative stress caused by solid Ehrlich tumor, probably due to the presence of antioxidant compounds, and had potential antineoplasic effect after a 30-day treatment.

A polysaccharide-rich fraction (ATF) of medicinal mushroom Agaricus brasiliensis was evaluated on the candidacidal activity, H2O2 and nitric oxide (NO) production, and expression of mannose receptors by murine peritoneal macrophages. Mice received three intraperitoneal (i.p.) injections of ATF and after 48 h their peritoneal resident macrophages were assayed against Candida albicans yeast forms. The treatment increased fungicidal activity and it was associated with higher levels of H2O2, whereas NO production was not affected. We also found that the treatment enhances mannose receptor expression by peritoneal macrophages, which are involved in the attachment and phagocytosis of non-opsonized microorganisms. Treatment of animals with ATF was able to enhance the clearance of C. albicans during the first 6 h after the experimental i.p. infection. Our results suggest that this extract can increase host resistance against some infectious agents through the stimulation of microbicidal activity of macrophages.