Anti-nociceptive and anti-inflammatory effects of the ethanol extract of Arenga pinnata (Wurmb) Merr. fruit.

ETHNOPHARMACOLOGICAL RELEVANCE: Arenga pinnata (Wurmb) Merr. is a medicinal and edible plant belonging to family Palmae. The fruits of this plant were used in traditional folk medicine due to its analgesia and anti-inflammatory activities. This study aimed to investigate the analgesic and anti-inflammatory properties and the mechanism of the ethanol extract of A. pinnata (Wurmb) Merr. fruit (EAF) on different experimental models. MATERIALS AND METHODS: High-performance liquid chromatography (HPLC) was used to determine the chromatographic profile and to analyze the composition of EAF. In the acute toxicity test, all mice were orally administered EAF at a maximum dosage of 26 g/kg and were then monitored for 14 days. The potential analgesic activity of EAF was evaluated by using animal pain models, namely the acetic acid-induced writhing test and the hot plate test in mice. The underlying mechanisms of analgesia were determined by pretreating with naloxone, capsaicin and cinnamaldehyde to evaluate the involvement of the opioid system and transient receptor potential channels (TRP channels). The anti-inflammatory activity of EAF was evaluated by using the following inflammatory animal models: xylene-induced ear edema in mice and Complete Freund's adjuvant (CFA)-induced paw swelling in rats. EAF was orally administered at the doses of 1.625, 3.25 and 6.5 g/kg in mice and 1.125, 2.25 and 4.5 g/kg in rats. The underlying mechanism of the anti-inflammatory activity was determined by enzyme-linked immunosorbent assay (ELISA) kits and real time-PCR used to measure the expression levels of interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), prostaglandin E2 (PGE2) and cyclooxygenase-2 (COX-2). Western blot analysis was used to determine the expression levels of proteins related to the nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPK) signaling pathways in paw tissues. RESULTS: Five compounds, namely (5-(hydroxymethyl) furan-2-yl) methanediol,4'-hydroxy-N-(4-hydroxy-3-methoxybenzoyl)-3',5'-dimethoxybenzamide, (+)-lyonirenisol-3a-O-ß-D-glucopyranoside, (-)-lyonirenisol-3a-O-ß-glucopyranoside and liquiritin, were firstly identified from A. pinnata (Wurmb) Merr. fruit by HPLC-UV analysis. In the acute toxicity test, no treatment-related toxicological signs or mortality was observed in mice administered doses up to 26 g/kg. Bodyweight was not obviously different among the treatment groups and the vehicle group. EAF significantly inhibited the pain response induced by acetic acid and increased the latency time in the hot plate test in mice. The anti-nociception effect of EAF in the formalin test was not alleviated by pretreatment with naloxone. However, the nociception induced by injection with capsaicin and cinnamaldehyde was significantly reduced by EAF. Compared with vehicle treatment, EAF significantly inhibited the formation of xylene-induced ear edema and CFA adjuvant-induced paw swelling. EAF markedly inhibited the production of IL-1ß, TNF-α, PGE2 and IL-6 induced by CFA in paw tissues. Furthermore, the phosphorylation of IKKα, IKKß, IκBα, p38, ERK1/2, and JNK and the nuclear translation of NF-κB p65 induced by CFA in paw tissues were significantly inhibited by EAF treatment compared with vehicle treatment. CONCLUSION: For the first time, this study provides pharmacological evidence for the analgesic and anti-inflammatory activities of EAF and the underlying mechanism, suggesting that EAF might be a potential candidate for reducing pain and inflammatory disorders.

Acute and subacute oral toxicity assessment of the oil extracted from Attalea phalerata Mart ex Spreng. pulp fruit in rats.

Attalea phalerata Mart. ex Spreng., popularly known as "bacuri", is a native plant from the brazilian Cerrado and used in folk medicine as a pulmonary decongestant, an anti-inflammatory for joints and antipyretic. There is an expectation about the use in chronic disease of the Attalea phalerata oil since its composition is high in carotenoids and beneficial fatty acids. The aim of the study was to evaluate the toxicological profile of the oil extracted from Attalea phalerata Mart. ex Spreng. pulp (APO). Acute and subacute toxicity studies were performed in male and female Wistar rats according to the OECD - Guidelines 425 and 407. For the acute toxicity, one single dose of the APO (2000mg/kg) was administered by gavage to five female rats. In the subacute toxicity, four different doses (125, 250, 500 and 1000mg/kg) of the APO were administered to male and female rats for 28 consecutive days. No deaths or behavioral changes were observed during both experiments as well as no changes in organ weights, hematological, histopathological parameters. The biochemical parameters showed changes in phosphatase alkaline and albumin levels, however these values are within the normal range for the species. A significant reduction in cholesterol and triglycerides was also observed in some of the animals treated with the APO. Therefore, the LD50 is higher than 2000mg/kg and the APO oil can be considered safe at the doses tested in rats. However, further assessments are required in order to proceed to clinical studies in humans.

Análise da citotoxicidade do extrato do fruto de juçara (Euterpe oleracea Mart) do Maranhão em células malignas humanas / Analysis of cytotoxicity of fruit extract juçara (euterpe oleracea mart) of Maranhão in human malignant cells

A juçara, Euterpe oleracea Mart., fruta indígena da Amazônia Legal, é rica em fitoquímicos com atividades anti-oxidante, antiinflamatória e anti-câncer. Este estudo tem por objetivo analisar os efeitos do extrato hidroalcoólico da casca, caroço e fruto total da juçara em diferentes linhagens de células malignas humana. Os frutos foram coletados no Parque da Juçara, localizado no Maracanã, município de São Luís, seguida da confecção da excicata que se mantém registrada no Herbário Rosa Mochel do Núcleo de Estudos Biológicos da Universidade Estadual do Maranhão. Os extratos hidroalcoólicos da casca, caroço e fruto total foram extraidos no Laboratório de Farmacologia e Psicobiologia da UERJ. As linhagens celulares utilizadas nos ensaios foram MCF-7 (adenocarcinoma de mama), CACO-2 e HT-20 (adenocarcinoma colo retal) e adenocarcinoma na mama (MDA-MB-468). As linhagens foram tratadas com 10, 20 e 40µg/mL dos extratos por 24 e 48 horas e feitas às análises. Células MCF-7 controle apresentaram núcleo proeminente com nucléolos evidentes. Após tratamento com o extrato hidroalcoólico da casca da juçara, as células mostraram morfologia arredondada com retração do citoplasma. O ensaio de viabilidade com MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) demonstrou uma redução na viabilidade das células. Após 48 horas, o tratamento das células com 20µg/mL do extrato da casca reduziu a viabilidade sendo que o efeito citotóxico do tratamento com 40µg/mL do extrato da casca foi potencializado. Células tratadas com 10µg/mL do extrato do caroço de juçara apresentavam-se arredondadas com consequente redução no volume celular. A concentração 20µg/mL de extrato hidroalcoólico do caroço, causou severa redução no volume das células e ocasionou o surgimento de vacúolos intracelulares. O mesmo foi observado após tratamento com 40µg/mL. O tratamento com 40µg/mL do extrato hidroalcoólico do fruto total, modificou drasticamente a morfologia das células MCF-7...

Juçara, Euterpe oleracea Mart., an indigenous fruit from Amazon, is rich in phytochemicals with antioxidant, anti-inflammatory and anti-cancer activity. This study aims to analyze the effects of the hydroalcoholic extract of the bark, seed and total fruit of juçara in different human malignant cell lines. Fruits were collected at the Maracana Ecological Park, in São Luís, followed by excicata manufacturing that remains registered in the Herbarium Rosa Mochel from the Nucleus of Biological Studies at the State University of Maranhão. The hydroalcoholic extracts of bark, seed and fruit were all obtained in the Laboratory of Pharmacology and Psychobiology UERJ. The cell lines used in the tests were MCF-7 and MDA-MB-468 (breast adenocarcinoma) and CACO-2 and HT-20 (colorectal adenocarcinoma). Strains were treated with 10, 20 and 40μg/mL of extracts for 24 and 48 hours. Control MCF-7 cells showed prominent nucleus with evident nucleoli. After treatment with the hydroalcoholic extract from the bark of juçara, the cells showed rounded morphology with retraction of the cytoplasm. The MTT viability assay showed a reduction in cell viability. After 48 hours, treatment of cells with 20μg/mL of bark extract reduced cell viability and the cytotoxic effect of treatment with 40μg/mL extract of the bark was potentiated. Cells treated with 10μg/mL of the bark extract were rounded with consequent reduction in cell volume. The concentration of 20μg/mL of bark extract caused severe reduction in volume of the cells and caused the appearance of intracellular vacuoles. The same was observed after treatment with 40μg/mL. Treatment with 40μg/mL of the hydroalcoholic extract of total fruit dramatically changed the morphology of the MCF-7 cells causing vacuolization and lysis with apparent loss of cytoplasmic contents. MTT assay showed a reduction in viability of MCF-7 cells treated with 20 and 40μg/mL after 24 hours of treatment. Analysis by electron microscopy showed the appearance...