Eugenia punicifolia (Kunth) DC, known as “pedra ume-caá” or “pitanguinha do cerrado”, is popularly used to treat infectious diseases like inflammation, and diabetes. Therefore, this study aimed to develop and validate an analytical method by high-performance liquid chromatography with diode array detector (HPLC-DAD) for the seasonal quantification of gallic acid (GA) and ellagic acid (EA), and to co-validate the method for quantification from the ethanol extract and fractions of the E. punicifolia leaves. Chromatographic separations were performed with a mobile phase gradient of acetonitrile/methanol/water acidified with 0.2% formic acid, a flow of 0.8 mL min 1, detection at 254 nm, and a C18 column (250 × 4.6 mm, 5 µm). Method validation parameters were determined following Brazilian legislation. The HPLC-DAD analytical method developed for the quantification of GA and EA was validated and demonstrated to be selective, linear, precise, accurate, robust, and without matrix effect. Accordingly, it is helpful for the analysis of these tannins in crude ethanol extracts and fractions from E. punicifoila leaves. Thus, our results may be of great help for quality control assessment and standardization of raw materials containing Eugenia punicifolia.

Abstract The antinociceptive and anti-inflammatory activities of crude ethanolic extract of Celtis iguanaea leaves and their active fractions are reported. The oral treatment with crude ethanolic extract (CEE; 100, 300 or 1000 mg/Kg) inhibited the number of writhings in a dose-dependent manner. The intermediate dose also inhibited formalin-induced nociception in both phases. The oral treatment with dichloromethane fraction (DF; 9 mg/Kg) produced antinociceptive effect in both phases of formalin test; however, the treatment with ethyl acetate fraction (EAF; 16 mg/Kg) reduced pain only in the second phase of this test. The oral treatments with CEE (300 mg/Kg) or DF (9 mg/Kg) reduced the nociception induced by capsaicin and pre-treatment with naloxone did not change these effects. The oral administration of CEE (300 mg/Kg), DF (9 mg/Kg) or ethyl EAF (16 mg/Kg) reduced ear edema, leukocytes migration and myeloperoxidase activity. Furthermore, the oral treatment with CEE (300 mg/Kg) or EAF (16 mg/Kg) reduced the level of Tumor Necrosis Factor - Alpha (TNF-α) in the pleurisy test. In conclusion, the DF showed antinociceptive activity that involves the vanilloid system as well as anti-inflammatory effect and the EAF showed anti-inflammatory activity involving the reduction of TNF-α cytokine.

ABSTRACT In this work, we developed and validated a HPLC-PDA method for the quantification of hibalactone in Hydrocotyle umbellata L., Araliaceae, subterraneous parts extracts and optimized its ultrasound-assisted extraction. Chromatographic separations were carried out with an isocratic mobile phase of acetonitrile/methanol/water (10:65:25), a flow of 0.8 ml min−1, detection at 290 nm and C18 column (250 × 4.6 mm, 5 µm). The method validation parameters were determined according to Brazilian legislation. The optimization of the hibalactone ultrasound-assisted extraction was performed using Box–Behnken design and response surface methodology. The HPLC method for hibalactone quantification proved to be selective, linear, precise, accurate and robust, being useful for the analysis of hibalactone in H. umbellata subterraneous parts extracts. The optimal ultrasound-assisted extraction conditions were obtained with solid-to-liquid ratio of 1:5 g ml−1, ethanolic strength of 70% (v/v) and temperature of 65 °C. The results can provide support of the quality control and standardization of raw materials from H. umbellata.

Levantamentos etnobotânicos de plantas nativas do cerrado evidenciam que as folhas deCeltis iguanaea (Jacq.) Sargent (Cannabaceae), popularmente conhecidas como “esporão-de-galo” são usadas na medicina popular para dores no corpo, asma, cólicas, má-digestão, infecções urinárias, disfunções renais, como estimulante ou como diurético. O presente trabalho teve como objetivo avaliar as possíveis atividades citotóxica, genotóxica e antigenotóxica do extrato aquoso das folhas de C. Iguanaea (CALE) através do teste do micronúcleo em medula óssea de camundongos. Para avaliar a genotoxicidade de CALE, os camundongos foram tratados por via oral com três diferentes concentrações de extrato (100, 300 e 500 mg.kg−1). Para a investigação da atividade antigenotóxica, foram utilizadas as mesmas doses concomitantemente a uma única dose i.p. de Mitomicina C (MMC, 4mg.kg−1). Com exceção do controle negativo (apenas 24h), todas as outras doses tiveram as frequências de eritrócitos policromáticos micronucleados (EPCMN) avaliadas em 24 e 48 horas após o tratamento. A Genotoxicidade foi avaliada por meio da frequência de eritrócitos policromáticos micronucleados (MNPCE), enquanto que a citotoxicidade foi mensurada pela razão entre eritrócitos policromáticos e normocromáticos (PCE / NCE). Os resultados mostraram que CALE não apresentou uma redução significativa na razão PCE/NCE, nem um aumento considerável na frequência de EPCMN. No entanto, CALE reduziu a toxicidade na medula óssea (aumento na razão PCE/NCE) e diminuiu a frequência de micronúcleos induzidos pela MMC. Nos resultados obtidos, CALE não apresentou efeitos citotóxicos e genotóxicos, contudo apresentou ações antigenotóxicas e anticitotóxicas nas condições experimentais aplicadas neste estudo.

Ethnobotanical surveys of Cerrado native plants show that leaves of Celtis iguanaea (Jacq.) Sargent (Cannabaceae), popularly known in Brazil as “esporão de galo”, are used in folk medicine for body pain, asthma, cramps, poor digestion, urinary infection, kidney dysfunctions, as well as a stimulant and diuretic. This work aimed at evaluating possible C. iguanaea aqueous leaf extract (CALE) cytotoxicity, genotoxicity, and antigenotoxicity using the mouse bone marrow micronucleous test. To assess CALE genotoxicity, Swiss mice were orally treated with three different extract concentrations (100, 300, and 500 mgkg−1). To evaluate its antigenotoxicity, the same doses were used simultaneously with a single i.p. dose of mitomycin C (MMC, 4mg.kg−1). The frequencies of micronucleated polychromatic erythrocytes (MNPCE) were evaluated 24 h and 48 h after administration except for the negative control (24 h). Genotoxicity was evaluated using the frequency of micronucleated polychromatic erythrocytes (MNPCE), whereas cytotoxicity was assessed by the polychromatic and normochromatic erythrocytes ratio (PCE/NCE). The results showed that CALE did not exhibit a significant reduction in the PCE/NCE ratio, neither a considerable increase in the frequency of MNPCE. Nonetheless, CALE reduced bone marrow toxicity (increased PCE/NCE ratio) and decreased the micronuclei frequency induced by MMC. We can conclude that CALE presented no cytotoxic and genotoxic effects, but showed antigenotoxic and anticytotoxic actions under the experimental conditions applied in this study.

A Hydrocotyle umbellata L., é uma espécie pertence à família Araliaceae, popularmente conhecida como acariçoba. Suas indicações na medicina popular incluem tratamento de úlceras cutâneas, eczemas e reumatismo. O objetivo deste trabalho é avaliar as atividades antinociceptiva e/ou anti-inflamatória do extrato etanólico das partes subterrâneas da acariçoba (EEA). O EEA reduziu a resposta nociceptiva do animais no teste das contorções abdominais induzidas por ácido acético e em ambas as fases do teste da formalina. O EEA também mostrou uma atividade analgésica supra-espinhal ao aumentar o limiar nociceptivo no método da placa quente. O EEA reduziu tanto o número de leucócitos migrados quanto o extravasamento plasmático para a cavidade pleural na pleurisia induzida por carragenina. Além de reduzir o edema de pata induzido por carragenina até a segunda hora e também o edema de pata induzido por dextrana. Em conclusão os nossos resultados mostram que o EEA de H. umbellata L. possui atividades analgésica e anti-inflamatória, e que um bloqueio da atividade ou redução da liberação de diferentes mediadores tais como histamina e serotonina, podem estar envolvidos nestes efeitos farmacológicos.

The Hydrocotyle umbellata L. is a specimen of the Araliaceae family popularly known as acariçoba. Its indications in folk medicine include treatment of skin ulcers, and rheumatism. The aim of this study was to evaluate the antinociceptive and anti-inflammatory activities of the ethanolic extract from acariçoba's underground parts (EEA). EEA reduced the nociceptive response of the animals as evaluated in the acetic acid-induced writhing test and in both phases of formalin test. EEA also presented a supraspinal analgesic activity by increasing the pain latency in the hot plate test. Moreover, EEA reduced the leukocytes migration and plasma extravasation to pleural cavity in the carrageenan-induced pleurisy, besides reducing the edema induced by carrageenan until the second hour and also the edema induced by dextran. In conclusion our results showed that EEA of H. umbellata L. presents analgesic and anti-inflammatory activities, and that a blockade of activity or reduction in the release of different mediators, such as histamine and serotonin, could be involved in these pharmacologic effects.

Acetic acid-induced writhing, hot-plate, carrageenan-induced pleurisy, formalin-induced pain, croton oil-induced ear edema, vascular permeability tests and phospholipase A2 activity assay were used to study the analgesic and/or anti-inflammatory activity of the hydromethanolic fraction of ethanolic extract from Spiranthera odoratissima A. St.-Hil., Rutaceae, leaves (HMF) and its subfraction (sub-Fr10-28). HMF and sub-Fr10-28 reduced the leukocyte migration on the carrageenan-induced pleurisy test; sub-Fr10-28 reduced the pain reaction time in the second phase of formalin-induced pain, as well as the ear edema and vascular permeability. Both HMF and sub-Fr10-28 inhibited the phospholipase A2 activity. These results suggest that the analgesic effect of this plant could be, in part, due to an anti-inflammatory action produced by the inhibition of phospholipase A2 activity.

The plant Hydrocotyle umbellata L., Araliaceae (water pennywort), is widely used in Brazilian folk medicine to reduce anxiety. This work investigates the anxiolytic-like effects of the ethanol extract from H. umbellata subterraneous parts as well as the extract's other putative central nervous system effects that could justify its common use. Oral dosing of the extract (0.3 and 1 g/kg) clearly showed an anxiolytic-like profile in the elevated plus maze test where it increased the percentage of entries into and the time spent in the open arms of the maze. In the marble-burying test, the extract induced anxiolytic-like effects only at a dose of 1 g/kg, which also causes mild sedative properties in other models. The sedated state was characterized by a slight reduction in spontaneous exploratory activity during the open field test and a potentiating of pentobarbital-induced hypnosis. No signs of motor impairment were detected in the rota rod or chimney tests. The extract did not show antidepressant properties in mice as assessed by the forced swimming test. These results support the use of H. umbellata in Brazilian folk medicine as an anxiolytic and contribute to the scientific knowledge of this possible phytotherapeutic resource.

This study evaluates the bioactivity of the crude ethanol extract and ethyl acetate, hexane and chloroform fractions obtained from Eugenia uniflora leaves using the hepatopancreas of Oreochromis niloticus L. as an experimental model. The ethanol extract and fractions were administered to the fish orally with their feed. Twenty-four hours later, the fish were sacrificed and their livers dissected, fixed in neutral formalin, embedded in paraffin and sectioned. Histological analyses were performed using Masson's trichrome and Haematoxylin-Eosin. Histochemical studies were performed using Feulgen, PAS (Periodic Acid Schiff) and PAS + salivary amylase and Sudan IV stain. The qualitative analysis of the material showed that the crude extract and the ethyl, chloroform and hexane fractions induced vasodilation, vascular congestion and toxicity due to the presence of eosinophilic granular cells, rodlet cells, some leukocytic infiltrate and rare focal necroses. The Nile tilapia proved to be a satisfactory model for screening plant products.

Os óleos essenciais de Toona ciliata, Cedrela odorata e C. fissilis foram analisados por CG-EM. Cedrela apresentou em maior percentagem sesquiterpenos formados a partir do precursor pirofosfato de cis- e trans-farnesila. Já Toona mostra uma tendência em produzir principalmente sesquiterpenos derivados do pirofosfato de trans-farnesila. Estes resultados confirmam que a classificação destes dois gêneros em uma mesma tribo, Cedreleae, continua problemática. As respostas em eletroantenogramas médios (EAGs) dos óleos essenciais de T. ciliata e C. odorata, em fêmeas de Hypsipyla grandella, foram significativamente maiores que aquelas obtidas em machos, sugerindo que as fêmeas utilizariam os odores destes óleos para a seleção da planta hospedeira (gêneros de Swietenioidea) e na escolha de locais para oviposição.

The essential oils of Toona ciliata, Cedrela odorata and C. fissilis have been analysed by GC-MS. Cedrela contains the main sesquiterpenes formed from the cis- and trans-farnesyl pyrophosphate. In contrast, Toona tend to produce mainly sesquiterpenes formed from the trans-precursor. These results show that the affiliation of Toona in the tribe Cedreleae together with Cedrela is still rather problematic. Mean electroantennogram responses (EAGs) to the essential oils from T. ciliata and C. odorata, in Hypsipyla grandella females were significantly greater than those obtained for males, suggesting that females would use attractant odours messages from the host-plant (genera of Swietenioideae) as search strategy behaviour for habitat location and oviposition.