Isovitexin as marker and bioactive compound in the antinociceptive activity of the Brazilian crude drug extracts of Echinodorus scaber and E. grandiflorus

Abstract Echinodorus scaber Rataj and Echinodorus grandiflorus (Cham. & Schltdl.) Micheli, Alismataceae, are popularly used to relieve inflammatory complaints and as diuretic. A study on the antinociceptive effect and selected marker compounds in eleven extracts from different locations was undertaken and their antinociceptive effect was assessed. The fingerprints were compared by HPLC-DAD and the content of vitexin, isovitexin, isoorientin and vitexin-2-O-rhamnoside were determined. All samples presented antinociceptive activity reducing the writhes by 36.4-62.5% and 47.4-79.8% at 10 and 50 mg/kg, respectively; indomethacin (5 mg/kg) reduced writhes by 82.6-90.1%. The content of the flavonoids C-glycosides, however, presented a strong variation. Isovitexin and isoorientin were found in all the samples, with content ranging from traces to 14.70 µg/mg and 2.12-84.27 µg/mg extract, respectively, while vitexin-2-O-rhamnoside occurred in quantifiable amounts only in 3 out of 11 samples ranging from 5.43 to 33.13 µg/mg extract; vitexin was not detected at all or detected in trace amounts. According to the fingerprints, the samples could be arranged in four main groups. All eleven extracts showed antinociceptive activity. Isovitexin was the only flavonoid present in all samples and can be regarded, acting in synergy with the other compounds or not, as the responsible for the antinociceptive activity. Therefore, isovitexin is a good choice as chemical marker when the antinociceptive activity of E. scaber and E. grandiflorus is investigated.

Hippocratic screening and subchronic oral toxicity assessments of the methanol extract of Vatairea macrocarpa heartwood in rodents

Vatairea macrocarpa (Benth.) Ducke, Fabaceae, is popularly known as 'angelim'. Its heartwood macerate is used to treat inflammation, gastric ulcer, diabetes and infections. The oral acute and subchronic toxicity of the methanol extract of V. macrocarpa heartwood (MEVm) was evaluated. In the Hippocratic screening, a single administration of MEVm was given orally to mice at doses ranging from 100 to 5000 mg/kg. In the subchronic study, MEVm was given orally as a daily administration for thirty days to Wistar rats at doses of 20, 100 and 500 mg/kg. In Hippocrtaic screening, doses of MEVm up to 5000 mg/kg did not cause any relevant behavioral changes or deaths thus making it impossible to establish the LD50. In subchronic assay, body weight gains and food intake were significantly reduced at the last week of treatment with 20 and 500 mg/kg dose. Serum triacylglycerides, total proteins and γ-glutamyltransferase activity were significantly reduced, while alkaline phosphatase activity was elevated. In hematological parameters, MEVm increased the percentage of segmented neutrophils cells at the highest dose. All alterations observed were minor in nature and were not accompanied by any relevant clinical signs or any histopathological changes. In conclusion, the results demonstrate relative safety profile of MEVm in the experimental animals.

Anti-inflammatory, antinociceptive, and antipyretic effects of methanol extract of Cariniana rubra stem bark in animal models

Cariniana rubra Miers (Lecythidaceae), popularly known as "jequitibá-vermelho'', is a large Brazilian tree whose bark is used in infusion and decoction for the treatment of inflammatory conditions. This study aims to assess the anti-inflammatory, antinociceptive, and antipyretic effects of Cariniana rubra methanolic stem bark extract (EM Cr) using experimental animals. Anti-inflammatory activity of EM Cr was tested on carrageenan and dextran-induced rat paw edema, carrageenan-induced pleurisy in rats and acetic acid-increase vascular permeability in mice. Antinociceptive and antipyretic activities were evaluated using acetic acid-induced writhing, formalin and hot-plate tests in mice, as well as brewer's yeast-induced pyrexia in rats. The extract inhibitied carrageenan and dextran-induced edema, reduced exudate volume and leukocyte migration on the carrageenan-induced pleurisy and on the vascular permeability increase induced by acetic acid. The EM Cr inhibited nociception on the acetic acid-induced writhing and in the second phase of formalin test, and decreased rectal temperature. It was, however, inactive against thermal nociception.Phytochemical analysis with EM Cr showed the occurrence of saponins, triterpenes, sterols and phenolic compounds. Phytosterols (β-sitosterol, stigmasterol), pentacyclic triterpenes (α- and β-amyrin as a mixture), arjunolic acid, a phytosterol glycoside (sitosterol 3-O-β-D-glucopyranoside), and triterpenoid saponins (28-β-glucopyranosyl-23-O-acetyl arjunolic acid; 3-O-β-glucopyranosyl arjunolic acid and 28-O-[α-L-Rhamnopyranosyl-(1→2)-β-glucopyranosyl]-23- O-acetyl arjunolic acid) were the main identified compounds. It can be presumed that EM Cr caused their effects by inhibiting the liberation and/or action of different inflammatory mediators. These findings support the traditional use of Cariniana rubra preparations to treat inflammation.

Cariniana rubra Miers (Lecythidaceae), popularmente conhecido como "jequitibá-vermelho'', é uma árvore brasileira de grande porte, cuja casca é utilizada nas formas de infusão e decocção para o tratamento de condições inflamatórias. Os efeitos antiinflamatório, antinociceptivo e antipirético do extrato metanólico da casca do caule de Cariniana rubra (EM Cr) foram avaliados em animais experimentais. A atividade antiinflamatória do EM Cr foi testada nos modelos de edema depata induzido por carragenina e dextrana em ratos, pleurisia induzida por carragenina em ratos e permeabilidade vascular aumentada por ácido acético em ratos. As atividades antinociceptiva e antipirética foram avaliadas utilizando os modelos de nocicepções induzidos por ácido acético e formalina, placa quente em camundongos e de pirexia, pela injeção de levedura de cerveja em ratos. O extrato inibiu o edema induzido porcarragenina e dextrana, reduziu o volume de exsudato e a migração de leucócitos na pleurisia induzida por carragenina eo aumento da permeabilidade vascular induzida por ácidoacético. O EM Cr inibiu a nocicepção nas contorções induzidas por ácido acético e na segunda fase do teste de formalina e diminuiu a temperatura retal. No entanto, foi inefetivo no teste da placa quente. A análise química por via úmida deu resultados positivos para saponinas, triterpenos, esteroides e compostos fenólicos. Fitosteróis e triterpenóides pentacíclicos (β-sitosterol, estigmasterol, α and β-amirinas em mistura e ácido arjunólico) e as saponinas triterpenoidais: 3-O-β-D-glucopiranosideo de sitosterol; ácido arjunólico 28-β-glucopiranosila-23-O-acetila; ácido arjunólico 3-O-β-glucopiranosila e ácido arjunólico 28-O-[α-L-rhamnopiranosil-(1→2)-β-D-glucopiranosila]-23-O-acetila. Pode-se presumir que os efeitos do EM Cr foram causados pela inibição da liberação e/ou ação de diversos mediadores inflamatórios. Estes resultados validam o uso tradicional das preparações caseiras de Cariniana rubra para tratar a inflamação.

Anti-inflammatory effect of the ethanolic extract from Bowdichia virgilioides H.B.K stem bark

Bowdichia virgilioides H.B.K stem bark (Fabaceae), locally known as "sucupira-preta", is a reputed folk-remedy to treat some inflammatory disorders. To validate its traditional claim, the ethanolic extract from B. virgilioides was evaluated in several animal models of inflammation and nociception. The extract at oral doses of 100 to 1000 mg/kg body weight caused a significant inhibition of carrageenan-induced hind paw oedema, suppression of exudate volume and leukocyte immigration in rat pleurisy induced by carrageenan, and reduction of granuloma weights in the model of subcutaneous granulomas promoted by cotton pellets. In addition, the plant extract significantly inhibited the vascular permeability increase induced by intraperitoneal acetic acid. It also showed marked antinociceptive effect in acetic acid-induced writhing test and in the second phase of formalin test in mice. These findings evidence the anti-inflammatory potential of Bowdichia virgilioides bark and supports its traditional use in inflammatory conditions.

A casca do caule de Bowdichia virgilioides H.B.K (Fabaceae), conhecida localmente como sucupira-preta, é um remédio popular muito utilizado para tratar inflamações. Com o objetivo de validar sua crença popular, o extrato etanólico de B. virgilioides foi avaliado em vários modelos experimentais de inflamação e nocicepção. O extrato administrado, via oral, em doses de 100 a 1000 mg/kg de peso corporal produziu inibição significativa no edema de pata induzido por carragenina, no aumento na permeabilidade vascular induzido por acido acético, no volume de exudato e na migração leucocitária no teste de pleurisia induzida por carragenina, bem como no peso de granulomas induzidos por pelotas de algodão, em ratos. Em camundongos, o EE Bv reduziu o número de contorções abdominais induzidas por ácido acético e a lambedura da pata na segunda fase do teste da formalina. Esses resultados validam o potencial anti-inflamatório da casca de Bowdichia virgilioides e referendam seu uso tradicional em condições inflamatórias.

Evaluation of the antifungal activity and mode of action of Lafoensia pacari A. St.-Hil., Lythraceae, stem-bark extracts, fractions and ellagic acid / Avaliação da atividade antifúngica e modo de ação dos extratos da entrecasca, frações e ácido elágico de Lafoensia pacari A. St.-Hil., Lythraceae

Stem-bark extracts, fractions and the isolated constituent, ellagic acid of Lafoensia pacari St. Hil. (Lythraceae) were in vitro assayed for antifungal activity against a panel of yeasts, hialohyphomycetes as well as dermatophytes with the microbroth dilution method. The EtOH extract and its fractions and ellagic acid exhibited activity against Candida spp and Saccharomyces cerevisiae with MIC values between 250-1000 µg/mL, but they showed no action against filamentous fungi and dermatophytes (MIC>1000 µg/mL). Active extracts were evaluated in Neurospora crassa hyphal growth inhibition and sorbitol assays and then the effect of ergosterol on the MIC of ellagic acid was studied. The active extracts and its fractions and ellagic acid showed a blotchy zone around the paper disk and induced malformations of the hypha. Besides, MIC of the ellagic acid against the Saccharomyces cerevisiae was raised from 62 to 250 µg/mL in the presence of sorbitol 0.8 M, suggesting that the ellagic acid would probably exert its action on fungal cell wall. These results indicate that ellagic acid might be the main active antifungal compound of Lafoensia pacari and further suggest that the mode of antifungal action of these extracts and ellagic acid could be associated with the inhibition of fungal cell wall.

Os extratos, frações e ácido elágico, isolados da entrecasca de Lafoensia pacari A. St.-Hil., Lythraceae, foram testados in vitro para atividade antifúngica, frente a um painel de leveduras, hialo-hifomicetos e dermatófitos, utilizando o método de microdiluição. O extrato EtOH, frações e ácido elágico exibiram atividade contra Candida spp. e Saccharomyces cerevisiae com valores de CIM entre 250-1000 µg/mL, porιm não mostraram ação contra fungos filamentosos e dermatófitos (CIM>1000 µg/mL). Os extratos ativos foram avaliados nos ensaios de inibição do crescimento das hifas de Neurospora crassa, no teste do sorbitol, e pelo estudo do efeito do ergosterol na CIM do ácido elágico. Os extratos ativos, frações e ácido elágico mostraram zonas manchadas ao redor dos discos de papel e induziram malformações nas hifas. Além disso, a CIM do ácido elágico contra Saccharomyces cerevisiae passou de 62 para 250 µg/mL na presença do sorbitol 0,8 M, sugerindo que o ácido elágico provavelmente poderia exercer ação na parede celular fúngica. Esses resultados indicam que o ácido elágico pode ser o principal composto antifúngico de Lafoensia pacari, sugerindo que o modo de ação antifúngico desses extratos e ácido elágico poderia estar associado à inibição da parede celular fúngica.

Triagem da atividade antibacteriana in vitro do látex e extratos de Croton urucurana Baillon / In vitro screening of antibacterial activity of the latex and extracts from Croton urucurana Baillon

Avaliou-se a atividade antibacteriana dos látex e extratos de diferentes polaridades e farmacógenos de Croton urucurana em ensaios antimicrobianos. Os farmacógenos foram coletados em Barão de Melgaço-MT. Os extratos foram obtidos por maceração a frio em hexano, diclorometano, acetato de etila, etanol e clorofórmio, rotaevaporado e seco em estufa. No ensaio de difusão em disco, os látex mostraram potente ação contra todas as bactérias, com exceção da E. coli, não diferindo quanto à potência e espectro antibacterianos. Extratos em hexano, diclorometano e etanol das folhas mostraram atividade contra S. pyogenes, K. pneumoniae, P. aeruginosa, S. typhimurium, S. aureus e S. epidermidis nas maiores doses. Extratos da entrecasca foram ativos contra S. aureus, S. epidermidis, P. aeruginosa, E. faecalis, S. pyogenes, E. coli, K. pneumoniae e S. typhimurium. Os látex apresentaram espectro de ação e potência maiores que os extratos obtidos da entrecasca e folhas. Dos extratos obtidos da entrecasca, o clorofórmico foi o mais potente, seguido pelo etanólico, indicando a presença de diferentes princípios ativos. Na microdiluição em caldo, os látex e extratos foram ativos, porém com maior potência para os primeiros. Os resultados evidenciam atividade antibacteriana para C. urucurana, em diferentes partes da planta e por diferentes metabólitos secundários.

We evaluated the antibacterial activity of the latex and extracts from different polarities and pharmacogens of Croton urucurana using antimicrobial assays. The pharmacogens were collected in Barão de Melgaço-MT. The extracts were obtained by cold maceration in hexane, dichloromethane, ethyl acetate, ethanol and chloroform. They were concentrated in rotatory evaporator and dried in stove. In the disk diffusion assay, the latex showed a potent action against all bacterial strains, excepting E. coli, not differing in the potency and antibacterial spectrum. The hexane, dichloromethane and ethanol extracts of leaves showed activity against S. pyogenes, K. pneumoniae, P. aeruginosa, S. typhimurium, S. aureus and S. epidermidis in the major doses. Extracts obtained of stem bark were actives against S. aureus, S. epidermidis, P. aeruginosa, E. faecalis, S. pyogenes, E. coli, K. pneumoniae and S. typhimurium. The latex showed higher potency and broad-spectrum of action than extracts from stem bark and leaves. Among the stem bark extracts, the chloroform was the most potent one, followed by the ethanol extract. This result suggests the presence of different active principles. In the broth-microdilution, the latex and all extracts showed activity, even though, the latex presented more potency. Our results indicate that C. urucurana presents antibacterial activity, in different parts of the plant and by different secondary metabolites.