Antioxidant properties of species from the Brazilian cerrado by different assays / Avaliação da propriedade antioxidante de espécies do cerrado no Brasil detectada por diferentes ensaios

The purpose of this study was to screen the antioxidant activity of medicinal plant extracts from the Brazilian cerrado, through other methods than the total phenolic content and its correlation with the antioxidant activity. Ethanolic extracts of ten species were evaluated through three antioxidant assays, in vitro, including 2,2-diphenyl-1-picrylhydrazyl (DPPH), total antioxidant activity and reducing power; and by using the Folin-Ciocalteu method the total phenolic content was determined. Ethanolic extracts of Stryphnodendron obovatum, Cecropia pachystachya and Duguetia furfuraceae showed strong antioxidant activity (IC50<5 µg mL-1) in the DPPH free radical scavenging assay; the species Vernonia phosphorea, Hymenaea stignocarpa and Jacaranda ulei may also be highlighted. These results were confirmed in the assays of total antioxidant capacity and reducing power. The extracts of S. obovatum and V. phosphorea showed an abundant phenolic content; therefore, the phenolic content may play a role in the antioxidant activity. These two species, traditionally used in Brazil, showed great power in these assay systems and may be a promising source for the development of natural antioxidants and future candidates for phytochemical and pharmacological studies in related diseases.

O objetivo desse trabalho foi triar a atividade antioxidante de extratos de plantas medicinais do cerrado do Brasil, por outros métodos além do conteúdo de fenóis totais e sua correlação com a atividade antioxidante. Assim, o extrato etanólico de dez espécies vegetais do cerrado brasileiro foi avaliado por três ensaios de atividade antioxidante, in vitro: 2,2-difenil-1-picrilhidrazil (DPPH); atividade antioxidante total e poder redutor; e o teor de fenóis determinado pelo reagente de Folin-Ciocalteu. O extrato etanólico de Stryphnodendron obovatum, Cecropia pachystachya e Duguetia furfuraceae apresentaram forte atividade antioxidante (CI50<5 mg mL-1) no ensaio com o DPPH, tendo destaque também as espécies Vernonia phosphorea, Hymenaea stignocarpa e Jacaranda ulei. Os extratos de S. obovatum e V. phosphorea demonstraram maiores teores de fenóis, indicando que esse grupo de substâncias possa ser a responsável pela atividade antioxidante. Essas duas espécies, usadas tradicionalmente no Brasil, representam fontes promissoras para o desenvolvimento de antioxidantes naturais e futuros estudos fitoquímicos e farmacológicos em doenças relacionadas.

Campomanesia lineatifolia Ruiz & Pav. as a gastroprotective agent.

ETHNOPHARMACOLOGICAL RELEVANCE: Campomanesia lineatifolia Ruiz and Pav. (Myrtaceae) is a native edible species found in the Amazon Rainforest, commonly known as gabiroba. In Brazil, Campomanesia species are frequently used in traditional medicine for gastrointestinal disorders. MATERIALS AND METHODS: The present study performed phytochemical analyses and determined both the in vitro antioxidant activity of the ethanolic extract of Campomanesia lineatifolia leaves (EEC) as well as its ethyl acetate fraction (EAFC). In this analysis, quercetin was used as a positive control. Gastroprotective activity was also investigated at different oral doses in two experimental models in rats - gastric lesion induced by ethanol and gastric lesion induced by indomethacin. In this analysis, cimetidine and sucralfate were used as positive controls. The area of gastric lesion underwent macroscopic and histomorphometric evaluations, while the mucus content was estimated by applying the periodic acid-Schiff stain. Oral acute toxicity was also assessed. RESULTS: Phytochemical studies revealed the presence of flavonoids and tannins. Catechin and quercitrin were isolated by bioguided chromatographic fractionation of EAFC. EEC and EAFC presented in vitro antioxidant activity. The oral administration of EEC and EAFC at doses 100-400 mg/kg (ethanol model) and at doses of 400-1200 mg/kg (indomethacin model) proved to be effective in preventing gastric ulcerations in rats. Pretreatment with EAFC (400mg/kg) significantly increased the gastric mucus content in the ethanol model. No animals died during the acute oral toxicology test. CONCLUSIONS: Results confirm the Brazilian ethnopharmacological use of Campomanesia lineatifolia as a gastroprotective agent and indicate that the anti-ulcer effect is most likely mediated by scavenging free radicals due to the polyphenol content and, at least in part, by increasing the mucus secretion and the mucosal defense. In addition, EEC and EAFC were found to be safe when applied to a 2000 mg/kg single oral dose.

Análise fitoquímica e atividade antimicobacteriana de extratos metanólicos de Jacaranda cuspidifolia Mart. (Bignoniaceae) / Phytochemical analysis and antimycobacterial activity of methanol extracts from Jacaranda cuspidifolia Mart. (Bignoniaceae)

Jacaranda cuspidifolia Mart., conhecida popularmente como "caroba", "jacarandá" ou "bolacheira", é utilizada medicinalmente para o tratamento da sífilis e da gonorréia. A atividade antimicobacteriana dessa espécie foi avaliada em ensaios in vitro com os extratos metanólicos das cascas e folhas, segundo o Método Analítico Alamar Blue (MABA). Os valores de concentração inibitória mínima para os extratos metanólicos das cascas e das folhas de J. cuspidifolia foram iguais a CIM = 250 μg mL-1 para ambos os extratos. A análise fitoquímica, por Cromatografia em Camada Delgada de gel de sílica, dos extratos metanólicos das cascas e folhas revelou a presença de taninos, flavonóides, terpenos, cumarinas e esteróides. A análise dos perfis dos extratos metanólicos por Cromatografia Líquida de Alta Eficiência de Fase Reversa registrou a presença de compostos fenólicos derivados do verbascosídeo sugerindo a provável responsabilidade pela ação antimicobacteriana.

Jacaranda cuspidifolia Mart., popularly known as "caroba", "jacaranda" or "bolacheira", is used as medicine for the treatment of syphilis and gonorrhea. The antimycobacterial activity of this species was assessed by means of in vitro assays with methanol extracts of barks and leaves according to the Microplate Alamar Blue Assay (MABA). The minimal inhibitory concentration values for methanol extracts of barks and leaves from J. cuspidifolia were MIC = 250 μg mL-1 for both extracts. Phytochemical analysis, by Thin Layer Chromatography on silica gel, of methanol extracts of barks and leaves revealed the presence of tannins, flavonoids, terpenes, cumarins and steroids. Analysis of the profiles of methanol extracts by High Performance Liquid Chromatography - Reversed Phase recorded the presence of phenolic compounds derivatives of verbascoside, suggesting their probable responsibility for the antimycobacterial action.

Antiviral activity of Distictella elongata (Vahl) Urb. (Bignoniaceae), a potentially useful source of anti-dengue drugs from the state of Minas Gerais, Brazil.

AIMS: To investigate the in vitro antiviral activity of Distictella elongata (Vahl) Urb. ethanol extracts from leaves (LEE), fruits (FEE), stems and their main components. METHODS AND RESULTS: The antiviral activity was evaluated against human herpesvirus type 1 (HSV-1), murine encephalomyocarditis virus (EMCV), vaccinia virus Western Reserve (VACV-WR) and dengue virus 2 (DENV-2) by the 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. LEE presented anti-HSV-1 [EC(50) 142.8 ± 5.3 µg ml(-1); selectivity index (SI) 2.0] and anti-DENV-2 activity (EC(50) 9.8 ± 1.3 µg ml(-1) ; SI 1.5). The pectolinarin (1) isolated from LEE was less active against HSV-1 and DENV-2. A mixture of the triterpenoids ursolic, pomolic and oleanolic acids was also obtained. Ursolic and oleanolic acids have shown antiviral activity against HSV-1. A mixture of pectolinarin (1) and acacetin-7-O-rutinoside (2) was isolated from FEE and has presented anti-DENV-2 activity (EC(50) 11.1 ± 1.6 µg ml(-1) ; SI > 45). Besides the antiviral activity, D. elongata has disclosed antioxidant effect. CONCLUSIONS: These data shows that D. elongata has antiviral activity mainly against HSV-1 and DENV-2, besides antioxidant activity. These effects might be principally attributed to flavonoids isolated. SIGNIFICANCE AND IMPACT OF THE STUDY: Distictella elongata might be considered a promising source of anti-dengue fever phytochemicals.

Chemosystematics of the rosiflorae.

The superorder Rosiflorae (sensu Dahlgren, 1980) belongs to the Angiospermae. It comprises twelve orders and thirty-eight families formed of species with varied habits widely distributed in temperate regions. The chemistry of Rosiflorae species is highly diversified; nevertheless it shows clearly phylogenetic affinity among the orders, except for Buxales. Flavonoids and triterpenoids are the real taxonomic markers for the superorder, due not only to the great number of occurrences, but also to the high structural diversity. On the other hand, the alkaloids are suitable as chemical markers only for the order Buxales. For orders and families of Rosiflorae, analysis of correlations among chemical parameters based on flavonoids and triterpenoids, with themselves and with the morphological and chemo-morphological parameters, showed evolutionary gradients among these taxa in which Trochodendrales occupy a primitive position while Saxifragales have the outpost. According to the types of flavonoids found in the superorder, there is clearly a higher incidence of flavonols than flavones, suggesting a primitive status of the Rosiflorae. Evolutionary advancement parameters relative to flavonoid hydroxyl protection show preferential protection mechanisms of glycosylation against methylation as well as a high percentage of free hydroxyl groups. The order Buxales has an isolated position in the superorder Rosiflorae with a high alkaloid production, which is quite exclusive to this taxon.

Chemosystematics of the Rosiflorae / Quimiossistemática de Rosiflorae

The superorder Rosiflorae (sensu Dahlgren, 1980) belongs to the Angiospermae. It comprises twelve orders and thirty-eight families formed of species with varied habits widely distributed in temperate regions. The chemistry of Rosiflorae species is highly diversified; nevertheless it shows clearly phylogenetic affinity among the orders, except for Buxales. Flavonoids and triterpenoids are the real taxonomic markers for the superorder, due not only to the great number of occurrences, but also to the high structural diversity. On the other hand, the alkaloids are suitable as chemical markers only for the order Buxales. For orders and families of Rosiflorae, analysis of correlations among chemical parameters based on flavonoids and triterpenoids, with themselves and with the morphological and chemo-morphological parameters, showed evolutionary gradients among these taxa in which Trochodendrales occupy a primitive position while Saxifragales have the outpost. According to the types of flavonoids found in the superorder, there is clearly a higher incidence of flavonols than flavones, suggesting a primitive status of the Rosiflorae. Evolutionary advancement parameters relative to flavonoid hydroxyl protection show preferential protection mechanisms of glycosylation against methylation as well as a high percentage of free hydroxyl groups. The order Buxales has an isolated position in the superorder Rosiflorae with a high alkaloid production, which is quite exclusive to this taxon.

A superordem Rosiflorae (sensu Dahlgren, 1980), Angiospermae, é composta por doze ordens e trinta e oito famílias. Em geral são plantas de hábito variado e muito freqüentes em regiões temperadas. A química das espécies de Rosiflorae é muito diversificada, mas evidencia a proximidade filogenética entre as ordens, com exceção de Buxales. Os flavonóides e os triterpenóides mostram-se como verdadeiros marcadores quimiossistemáticos em nível de superordem, devido, não somente ao seu grande número de ocorrências, mas também pela sua elevada diversidade estrutural; contudo os alcalóides são os marcadores para Buxales. Avaliação das correlações dos parâmetros químicos entre si e com os parâmetros morfológico e químico-morfológico, para ordens e famílias de Rosiflorae, com base nos seus flavonóides e triterpenóides, evidenciaram uma grande proximidade filogenética entre esses táxons, além de mostrar gradientes evolutivos, em que Trochodendrales e Saxifragales são posicionadas como a ordem mais primitiva e a ordem mais evoluída, respectivamente. Com relação aos tipos flavonoídicos produzidos na superordem, verifica-se uma maior produção de flavonóis em relação às flavonas, o que acarreta uma baixa relação favona/flavonol, confirmando o posicionamento primitivo da superordem, indicado pelos índices morfológico e químico-morfológico. A proteção das hidroxilas flavonoídicas mostra uma nítida preferência na proteção por glicosilação e desproteção em detrimento à proteção por metilação. A ordem Buxales praticamente encontra-se isolada na superordem com grande produção de alcalóides muito característicos, sendo bastante diferenciada da produção alcaloídica relativamente pobre da superordem.

Avaliação das atividades antioxidante, antimicrobiana e citotóxica de Couepia grandiflora Benth. (Chrysobalanaceae) / Evaluation of cytotoxicity, antioxidant, antibacterial and antifungal activities of Couepia grandiflora Benth. (Chrysobalanaceae)

Couepia grandiflora Benth. é conhecida popularmente como fruta-de-ema, pertence à família Chrysobalanaceae, e ocorre em regiões tropicais e subtropicais. Plantas dessa família são utilizadas pelas populações, principalmente, para o tratamento da diarréia, disenteria e malária. Neste trabalho, avaliou-se as atividades citotóxica, antibacteriana, antifúngica e antioxidante dos extratos hexânico e etanólico de C. grandiflora. O teste com Artemia salina usado para avaliar a citotoxicidade dos extratos hexânico e etanálico apresentou valores de CL50 de 0,1082 mg/mL e 0,1408 mg/mL, respectivamente. O extrato etanálico de C. grandiflora mostrou atividade antibacteriana contra Staphylococcus aureus e Pseudomonas aeruginosa, enquanto que o extrato hexânico mostrou atividade somente contra P aeruginosa. Os dois extratos não mostraram atividade contra Escherichia coli. As Concentrações Inibitórias Mínimas (CIM) do extrato etanólico foram de 188, 125, 188 μL/mL, para Staphylococcus aureus, Escherichia coli e Pseudomonas aeruginosa, respectivamente. Quanto à avaliação da atividade antioxidante, os extratos hexânico e etanólico mostraram-se ativos com valores de CE50= 3,1 e 5,6 μg/mL, respectivamente, quando comparados com os padrões. Na avaliação da atividade antifúngica, os extratos etanálico e hexânico de C. grandiflora não apresentaram tal atividade. Conclui-se que os extratos etanólico e hexânico de C. grandiflora demostraram atividades antibacteriana, antioxidante e citotoxicidade frente à Artemia salina e ausência de atividade antifúngica.

Couepia grandiflora Benth., known popularly as ema fruit, belongs to the family Chrysobalanaceae, and is distributed in tropical and subtropical areas. Plants of this family are used mainly for the treatrnent of the diarrhea. dysentery and malaria. In this work, it was evaluated the cytotoxic, antibacterial, antifungal and antioxidant activíties of the hexane and ethanolic extracts from C. grandiflora. The cytotoxic activíty of the hexane and ethanolic extracts showed a LC50 of 0.1082 mg/mL and 0.1408 mg/mL, respectively. The ethanolic extract of C. grandiflora showed antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa. while the hexane extract showed activity just against P. aeruginosa. The two extracts did not show activity against Escherichia coli. The Minímum Inhibitory Concentrations (MIC) of the ethanolic extract were 188, 125 and 188 μL/mL for Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, respectively. The evaluation of the antioxidant activity of the hexane and ethanolic extracts showed good results when compared to the reference drugs(EC50=3.1 and 5.6 mg/mL, respectively). It was not detected antifungal activity for hexane and ethanolic extracts from C. grandiflora.

Avaliação de atividades biológicas das sementes de Stryphnodendron obovatum Benth. (Leguminosae) / Evaluation of biological activities of the seeds of Stryphnodendron obovatum Benth. (Leguminosae)

Stryphnodendron obovatum Benth., conhecido como "barbatimão", é uma espécie pertencente à família Leguminosae, sub-família Mimosoideae, e é amplamente distribuído em campos e cerrados. Na medicina popular, cascas de S. obovatum são usadas no tratamento de processos inflamatórios, como cicatrizante, para diarréia, frieira. Neste trabalho investigou-se a presença de proteínas e as atividades citotóxica, antibacteriana, antifúngica do extrato salino das sementes de S. obovatum. O extrato salino S. obovatum não apresentou toxidade frente ao ensaio com Artemia salina, nem mostrou atividade antibacteriana contra Staphylococcus aureus, Pseudomonas aeruginosa e Escherichia coli. Na avaliação da atividade antioxidante, o extrato salino apresentou uma CE50 de 12, 193 µg/mL, enquanto a do padrão positivo BHT foi 2,98 µg/mL. O extrato salino de S. obovatum não apresentou atividade antifúngica, tanto na técnica de bioautografia com o fungo Cladosporium sphaerospermum, quanto no método de difusão em disco, realizado com Candida albicans. Foi realizado teste de atividade enzimática na qual observou-se a hidrólise do substrato H-D-Benzoil-arginina-p-nitroanilida (Bz-Arg-pNan).

Stryphnodendron obovatum Benth., popularly known as "barbatimão", belongs to the Leguminosae fami/y, of the Mimosoideae subfamily, and is present in fields and in "cerrados". S. obovatum bark is used in popular medicine for treating inflammatory processes, for healing wounds, and as cure for diarrhea and chílblain. This research investigates the presence of proteins and the cytotoxic, antifungal, antibacterial and antioxidant activities of the S. obovatum seed saline extract. The saline extract did not show cytotoxicity against Artemia salina nor any antibacterial activíty against Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia colí. The evaluation of the antioxidant activity showed a CEso=12.193 µg/mL, and the BHT positive pattern presented 2.98 µg/mL. The S. obovatum saline extract was tested against Cladosporium sphaerospermum and Candida albicans, using the bioautography technique and the disk diffusion method. Benzoyl­arginine-p-nitroanilide (Bz-Arg-pNan) was hydrolyzed by the saline extract.

High-speed countercurrent chromatography as a valuable tool to isolate C-glycosylflavones from Cecropia lyratiloba Miquel.

A new apigeninglycoside, apigenin 6-C-galactosyl-6"-O-beta-galactopyranoside (1), isoorientin, and a mixture of orientin and isovitexin were isolated from leaves of Cecropia lyratiloba by high-speed countercurrent chromatography using a solvent system containing ethyl acetate, butanol, methanol and water. The structural elucidation of 1 was based on NMR spectroscopy.