Chemical characterization of Brazilian savannah Byrsonima species (muricis) and their impact on genomic instability and chemopreventive effects.

The identification of new drugs with few or no adverse effects is of great interest worldwide. In cancer therapy, natural products have been used as chemopreventive and chemotherapeutic agents. Plants from the Brazilian savannah belonging to the Byrsonima genus are popularly known as muricis and have attracted much attention due to their various pharmacological activities. However, there are currently no data on these plants concerning their use as chemopreventive or chemotherapeutic agents in human cell lines. The present study assessed the potential of B. correifolia, B. verbascifolia, B. crassifolia, and B. intermedia extracts as natural alternatives in the prevention and/or treatment of cancer. The chemical constituents present in each extract were analyzed by electrospray ionization-mass spectrometry (ESI-MSN). The mutagenic/antimutagenic (micronucleus assay), genotoxic/antigenotoxic (comet assay), apoptotic/necrotic (acridine orange/ethidium bromide uptake), and oxidative/antioxidative (CM-H2DCFDA) effects of the extracts and their influence on gene expression (RTqPCR) were investigated in nonmetabolizing gastric (MNP01) and metabolizing hepatocarcinoma (HepG2) epithelial cells to evaluate the effects of metabolism on the biological activities of the extracts. The genotoxicity, mutagenicity, and apoptotic effects observed in HepG2 cells with B. correifolia and B. verbascifolia extracts are probably associated with the presence of proanthocyanidins and amentoflavone. In MNP01 cells, none of the four extracts showed mutagenic effects. B. crassifolia and B. intermedia extracts exhibited strong antimutagenicity and enhanced detoxification in HepG2 cells and antioxidant capacities in both types of cells, possibly due to the presence of gallic and quinic acids, which possess chemopreventive properties. This study identifies for the first time B. correifolia and B. verbascifolia extracts as potential agents against hepatocarcinoma and B. crassifolia and B. intermedia extracts as putative chemopreventive agents.

Mutagenicity of two species of the genus Alchornea measured by Salmonella microsome assay and micronucleus test / Mutagenicidade de duas espécies do gênero Alchornea avaliadas através de ensaios com Salmonella microssomo e teste do micronúcleo

Some species of the plant genus Alchornea (family Euphorbiaceae) are widely used in popular medicine, mainly in South America and in Africa. Several kinds of biological activity have been seen in the species: antioxidant, antifungal, anti-inflammatory, antibacterial, cytotoxic against tumor cell lines and inhibitory to the replication of HIV-1 and HIV-2. In Brazil, the species Alchornea castaneaefolia Willd. A. Juss. and Alchornea glandulosa Poepp. & Endl. are used by the local population to treat rheumatism, arthritis and muscular pains. In view of the popular use of these plants as medicines and the potential risks from their consumption, we assessed the mutagenic potential of chloroform and methanol extracts of the leaves of these plant species, employing the in vivo micronucleus test and the Ames assay. The data obtained showed that the chloroform extracts were not mutagenic. The methanol extract of A. castaneaefolia was mutagenic to strain TA98 of Salmonella typhimurium and the methanol extract of A. glandulosa to strains TA98 and TA97a. The methanol extracts of both species of Alchornea were mutagenic in vivo at the largest dose employed. The probable mutagenic agents involved were the aglycone quercetin and amentoflavone, present in both species.

Algumas espécies de plantas do gênero Alchornea (Euphorbiaceae) são conhecidas por apresentarem as atividades biológicas: antioxidante, antifúngica, antiinflamatória, antibacteriana, citotóxica para células tumorais e inibidoras da replicação dos vírus HIV-1 e HIV-2. São também amplamente usadas na medicina popular na America do Sul e África. No Brasil, Alchornea castaneaefolia Willd. A. Juss. e Alchornea glandulosa Poepp. & Endl. são usadas para tratamento do reumatismo, artrite e dores musculares. Devido ao uso medicinal dessas plantas e o potencial risco do seu consumo indiscriminado, no presente trabalho foi avaliada a atividade mutagênica dos extratos metanólico e clorofórmico das folhas, empregando o teste do micronúcleo in vivo e o teste de Ames. Os resultados mostraram que o extrato clorofórmico não apresentou mutagenicidade, porém, o extrato metanólico de A. castaneaefolia foi mutagênico para a linhagem TA98 de Salmonella typhimurium e o extrato metanólico de A. glandulosa para as linhagens TA98 e TA97a. O extrato metanólico de ambas as espécies também apresentaram mutagenicidade positiva nos ensaios in vivo na maior concentração usada. Os prováveis agentes mutagênicos envolvidos foram a quercetina aglicona e amentoflavona presentes em ambas as espécies.

In vivo assessment of DNA damage and protective effects of extracts from Miconia species using the comet assay and micronucleus test.

The genus Miconia comprises approximately 1000 species belonging to the Melastomataceae family. Several crude plant extracts from Miconia and their isolated compounds have shown biological activities, such as analgesic and anti-neoplastic action; however, no studies concerning their effects on DNA are available. The present study aimed to evaluate, in vivo, the genotoxic and mutagenic effects of four species of plants from Miconia genus using the comet assay and micronucleus test. Their possible protective effects were also evaluated in experiments associating the plant extracts with cyclophosphamide (CPA). The methanolic extracts of Miconia albicans, Miconia cabucu, Miconia rubiginosa, Miconia stenostachya and the chloroformic extract of M. albicans were investigated. For genotoxic and mutagenic evaluations, three concentrations were tested, 200, 400 and 540 mg/kg body weight (bw), based on the solubility limit of the extract in distilled water. For the protective effects, only the highest dose was evaluated against 40 mg/kg bw of CPA. Blood was removed from mice tails pre- (T0) and post-treatment (T1-30 h) for the micronucleus test and 24 h post-treatment for the comet assay. The Student's t-test was used to compare data obtained at T0 and T1, the analysis of variance-Tukey test was used to compare between groups in the micronucleus test and the Kruskal-Wallis and Dunn's test were used to compare different groups in the comet assay. All the extracts induced alterations in DNA migration (comet assay); however, no mutagenic effect was observed in the micronucleus assay. All extracts showed a protective effect against CPA in both assays. Our study showed that the use of crude extracts could be more advantageous than the use of isolated compounds. The interaction between phytochemicals in the extracts showed efficacy in reducing mutagenicity and improving the protective effects.

Avaliação in vivo da anticlastogenicidade de extratos de plantas medicinais do gênero Miconia através do teste do micronúcleo / In vivo evaluation of anticlastogenicity of extracts from medicinal

O gênero Miconia possui aproximadamente 1000 espécies, e para algumas, já foram descritas atividades biológicas como a analgésica e antimicrobiana. Esse trabalho teve como objetivo avaliar os possíveis efeitos protetores e citotóxicos dos extratos metanólicos de M. albicans, M. cabucu, M. rubiginosa e M. stenostachya e do extrato clorofórmico de M. albicans em células da medula óssea de camundongos na dose de 540 mg/kg p.c. Os extratos foram administrados via gavage e a ciclofosfamida (CPA) foi aplicada intraperitonealmente 1h, após a suplementação com os extratos. Todos os animais foram submetidos à eutanásia 30h após o tratamento. As células analisadas foram retiradas da medula óssea de acordo com protocolo descrito por Schmid (1975). A citotoxicidade dos extratos foi avaliada pela percentagem de eritrócitos policromáticos (PCE) em 200 eritrócitos (PCE + NCE). Foram analisados 2000 PCEs por animal e anotadas as freqüências de MNPCEs. Os resultados obtidos mostraram que nenhum dos extratos associados à CPA apresentou efeito citotóxico e somente os extratos de M. rubiginosa, M. stenostachya mostraram efeito protetor ao DNA. A análise química dos extratos mostrou que as quatro espécies estudadas contêm, principalmente, flavonóides, compostos fenólicos e taninos. A caracterização fitoquímica desses extratos poderia contribuir para elucidação do efeito protetor apresentado somente pelas espécies M. rubiginosa e M. stenostachya, além de possibilitar o estudo de outras possíveis atividades terapêuticas.

The genus Miconia is comprised of approximately 1000 species. For some of them, biological activitieshave already been described such as the analgesic and the anti-microbial ones. The purpose of this workwas to evaluate the possible protective and cytotoxic effects of the methanolic extract from M. albicans,M. cabucu, M. rubiginosa and M. stenostachya and the chloroformic extract from M. albicans in micebone marrow cells in 540 mg/kg p.c. dose. The extracts were administered by means of forced feedingand the cyclophosphamide (CPA) was applied intraperitonially one hour after supplementation withextracts. All animals were submitted to euthanasia 30 hours after the treatment. The analyzed cells wereextracted from mice bone marrow according to protocol described by Schmid (1975). The cytotoxicityof the extracts was evaluated through the percentage of polychromatic erythrocytes (PCE) in 200erythrocytes (PCE + NCE). Two thousand PCEs of each animal were analyzed and the micronucleatedpolychromatic erythrocytes (MNPCEs) frequencies were scored. The results obtained indicated that

Absence of mutagenicity in somatic and germ cells of mice submitted to subchronic treatment with an extract of Croton cajucara Benth. ( Euphorbiaceae )

The plant Croton cajucara Benth. (Euphorbiaceae) is a medicinal plant from the Brazilian Amazon where it is commonly known as sacaca. The principal compound isolated from C. cajucara stem-bark extracts is the clerodane-type diterpene trans-dehydrocrotonin (DCTN) which presents several biological activities, including antiulcerogenic, anti-inflammatory, hypoglycemic, antimutagenic and antitumoral activity. However, few studies have been carried out to evaluate the therapeutic potential of raw C. cajucara extracts. We studied mutagenicity and antimutagenicity effects of C. cajucara methanol extract using the micronucleus assay in bone marrow cells and the dominant lethal assay in mice submitted to subchronic treatments. The blood testosterone levels of the mice were also measured to assess the effects of the methanol extract on testes function. Statistical analysis of the data obtained in this study showed no statistically significant mutagenicity attributable to C. cajucara stem-bark extracts, nor did such extracts show antimutagenic activity at the concentrations assessed. The testosterone concentration was normal in all the mice studied.