Sambucus nigra: A traditional medicine effective in reducing inflammation in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Sambucus nigra L. is a plant of European origin and popularly known as elder, elderberry, black elder, European elder, European elderberry, and European black elderberry, being described in pharmacopoeia of several countries. Its flowers and berries have been used in folk medicine to treat feverish conditions, coughing, nasal congestion, and influenza besides its popular use as anti-inflammatory, analgesic, and diuretic agent. AIM OF THE STUDY: The aim of this investigation was to elucidate the anti-inflammatory and the relaxant effect of the lyophilized aqueous extract obtained from S. nigra's flowers on in vivo and in vitro inflammation assays and on the isolated rat vascular and airway smooth muscle tissue. MATERIAL AND METHODS: The anti-inflammatory activity of the extract was investigated using carrageenan-induced inflammation model in the subcutaneous tissue of male Swiss mice orally treated with S. nigra extract (30, 100, 300 or 600 mg/kg). Leukocyte influx and the secretion of chemical mediators were quantified in the inflamed exudate. Additionally, histological analysis of the pouches was performed. N-Formyl-methionine-leucine-phenylalanine-induced chemotaxis, lipopolysaccharide-induced TNF, IL-6, IL-1ß, IL-10 and NO production, and adhesion molecule expression (CD62L, CD49d and CD18, flow cytometry) were analyzed in vitro using oyster glycogen-recruited peritoneal neutrophils or macrophages (RAW 264.7) stimulated with LPS and treated with the extract (1, 10 or 100 µg/mL). The resolution of inflammation was accessed by efferocytosis assay, and the antinociceptive activity was investigated using carrageenan-induced mechanical hypersensitivity. Finally, the effect of the extract was evaluated in isolated rat aorta and trachea rings. RESULTS: The oral treatment with S. nigra promoted reduction in the neutrophil migration as well as the decrease of TNF, IL-1ß and IL-6 levels in the inflamed exudate. In vitro treatment with S. nigra decreased NO2-, TNF, IL-1ß and IL-6 and promoted increase of IL-10 in LPS-stimulated neutrophils. Similarly, the extract reduced the NO2-, TNF and IL-6 in LPS-stimulated macrophages. Rutin, the major constituent of S. nigra extract reduced NO2-, TNF, IL-1ß, and IL-6 and promoted the increase of IL-10 in LPS-stimulated neutrophils supernatant. The extract also shed CD62L and CD18 expressions. The extract was able to increase the efferocytosis of apoptotic neutrophils by increasing the IL-10 and decreasing the TNF levels. Additionally, the extract reduced the hypersensitivity induced by carrageenan and promoted a relaxant effect in isolated vascular and non-vascular rat tissue. CONCLUSIONS: S. nigra flowers extract presents anti-inflammatory effect by modulating macrophage and neutrophil functions including the production of inflammatory mediators and cell migration, by promoting efferocytosis and consequently the resolution of acute inflammation, besides exerting antinociceptive effects, scientifically proving its popular use as medicinal plant. Allied to the relaxant effect in both vascular and non-vascular smooth muscle tissue, S. nigra extract represents an important tool for the management of acute inflammation.

RP-HPLC and LC-MS-MS determination of a bioactive artefact from Ipomoea pes-caprae extract

Abstract Solvents play important and critical role in natural product chemistry and could generate artefacts during the extraction and purification of metabolites from a biological matrix. This study aimed to correlate the chromatographic profile with biological activity of Ipomoea pes-caprae (L.) R. Br., Convolvulaceae, extracts obtained with hydroethanolic extraction. Thus, aerial parts of I. pes-caprae were extracted with different concentration of ethanol (50, 70 and 90°GL) and the obtained extracts were analysed by HPLC-UV. HPLC data were studied employing chemometrics to discriminate the samples. Moreover these samples were further characterized by using UPLC-QTOF/MS data. The extracts were also biomonitored through the paw-oedema and spontaneous nociception induced by trypsin in mice. Different chromatographic profiles were obtained and the exploratory analysis clearly revealed higher level of ethyl caffeate in extracts of lower strength of ethanol (50°GL). This compound was suggested to be an artefact formed by transesterification of caffeoylquinic acid derivatives present in the plant, once it was not observed when other solvents were employed. During the biological assay, only the extract obtained with ethanol 50°GL presented significant inhibition of inflammation (45 ± 9%) and nociception (24 ± 3%). Ethyl caffeate seems to be linked to the anti-inflammatory effect since it reduced 86 ± 5% of paw-oedema induced by trypsin. Artefacts could contribute to the biological activity of herbal preparations and consequently lead to misinterpretation of the results.

Photochemoprotective effects against UVA and UVB irradiation and photosafety assessment of Litchi chinensis leaves extract.

Actually, there has been an increase in the use of natural products as skin photoprotective agents. In this way, the aim of present study was to investigate the L. chinensis leaves extract photochemoprotection potential and photosafety using in vitro methods. The extract cytotoxicity, cytoprotection and photochemoprotection against UVA and UVB radiation were assayed in L929 cells. The DNA damage was evaluated by comet assay. DCFH-DA, SOD, CAT and GSH assays were employed to verify the oxidative stress. We also determined the spectrophotometric Solar Protection Factor (SPF) of the extract. The photosafety was evaluated in red blood cells (RBC). In addition, the HET-CAM and agarose overlay tests were employed to evaluate the irritant potential. The results obtained show that extract is not cytotoxic and present cytoprotective activity against H2O2 and is able to protect the cells against DNA damage induced by UVA and UVB radiation. The extract was able to reduce the ROS production. The SPF obtained was 18.9 at 1mg/mL. We demonstrate that L. chinensis extract is photosafe and protect RBCs against oxidative damage, and did not induce irritation. Data herein obtained pointed out the potential of L. chinensis extract for photochemoprotection against UVA/UVB radiation and its damaging effects on human skin.

Gastroprotective activity of methanol extract and marrubiin obtained from leaves of Marrubium vulgare L. (Lamiaceae).

OBJECTIVES: The purpose of this study was to assess the gastroprotective properties of the methanol extract and the diterpene marrubiin obtained from the leaves of M. vulgare. METHODS: Assays were performed using different protocols in mice. Studies focusing on mechanisms of gastroprotection were also undertaken. KEY FINDINGS: In the model of ethanol-induced ulcers, we observed a significant reduction in all the parameters analysed; the curative ratios obtained were 49.31±0.57, 74.31±0.91 and 79.86±0.59 for the groups treated with 50 and 100mg/kg of extract of M. vulgare and omeprazole (30mg/kg), respectively. For indomethacin-induced ulcers, the percentages of ulcer inhibition were 50.32±5.60, 66.24±4.30, 82.17±04.09 and 67.52±4.38, for the groups treated with 25, 50 and 100mg/kg M. vulgare and positive control (cimetidine), respectively. In both models, the marrubiin (25mg/kg) produced a significant reduction in all the parameters when compared with the control group (P<0.01). There was also a significant increase in pH and mucus production in the groups treated with M. vulgare extract and marubiin. The results also demonstrated that the gastroprotection induced by the extract and marubiin is related to the activity of nitric oxide and endogenous sulfhydryls, which are important gastroprotective factors. CONCLUSIONS: The results of this study show that the extract of M. vulgare and marrubiin displays antiulcer activity and that this effect can be partly attributed to the isolated diterpene.