Biotransformation of a tetrahydrofuran lignan by the endophytic fungus Phomopsis Sp.

The biotrasformation of the tetrahydrofuran lignan, (-)-grandisin, by the endophitic fungus Phomopsis sp, obtained from Viguiera arenaria, led to the formation of a new compound determined as 3,4-dimethyl-2-(4'-hydroxy-3',5'-dimethoxyphenyl)-5-methoxy-tetrahydrofuran. The metabolite was evaluated against the parasite Trypanosoma cruzi, the causative agent of Chagas's disease, and showed a trypanocidal activity (IC50 9.8 μmol L-1) similar to the natural precursor (IC50 3.7 μmol L-1).A biotransformação da lignana tetraidrofurânica, (-)-grandisina, pelo fungo endofítico Phomopsis sp, obtido de Viguiera arenaria, conduziu à formação de um novo metabólito caracterizado como 3,4-dimetil-2-(4'-hidróxi-3',5'-dimetóxifenil)-5-metóxi-tetraidrofurano. O metabólito foi analisado contra o parasita Trypanosoma cruzi, o agente causador da doença de Chagas, e mostrou uma atividade tripanocida (IC50 9,8 μmol L-1) similar ao precursor natural (IC50 3,7 μmol L-1).Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Antinociceptivna i protuupalna svojstva vodeno-etanolnog ekstrakta pokožice grožđa vrste Vitis labrusca izolirane iz otpada vinske industrije

Research background. Extracts from grape pomace, including the wine, show many biological effects such as antioxidant and anti-inflammatory activities. Unfortunately, winemakers discard the bagasse, so the waste is not exploited, although it contains bioactive compounds with antioxidant and anti-inflammatory properties. The work aims to analyze the hydroethanolic extract of peels from Vitis labrusca agro-industrial waste and to evaluate its antinociceptive and anti-inflammatory properties. This study is relevant for reusing a residue and adding value to the grape economic chain. Experimental approach. A representative sample of pomace was obtained and the peels were used to produce the extract. The phenolic compounds were determined by mass spectrometry in multiple reaction monitoring mode and Folin-Ciocalteu colorimetric method, using gallic acid as standard. The biological analyses were carried out using mice orally treated with crude extract at doses of 30, 100 and 300 mg/kg. We evaluated mechanical hyperalgesia by the von Frey method, thermal heat hyperalgesia using a hot plate at 55 °C, paw edema using a pachymeter, and neutrophil recruitment by measurement of myeloperoxidase activity. The nephrotoxicity and hepatotoxicity were evaluated by biochemical analyses using blood samples that were collected after the Vitis labrusca administration. Results and conclusions. In all wet winemaking residues peel mass fraction was 75%, and in dry residues 59%. We identified nine anthocyanins (3-O-glucosides: peonidin, delphinidin, petunidin and malvidin; 3-p-coumaroyl-glucosides: cyanidin, peonidin, petunidin and malvidin, and malvidin-3,5-diglucoside), five flavonoids (apigenin-7-glucoside, luteolin-7-glucoside, quercetin-3-galactoside, isorhamnetin-3-glucoside and myricetin-3-rutinoside), and mass fraction of phenolic compounds, expressed as gallic acid equivalents, was 26.62 mg/g. In vivo assays showed that Vitis labrusca extract at mass fractions 100 and 300 mg/kg reduced carrageenan-induced mechanical and thermal hyperalgesia, 50% of the paw edema, and neutrophil recruitment. In addition, there were no indications of nephrotoxicity and hepatotoxicity. Our extract obtained from winemaking residue has analgesic and anti-inflammatory properties, related at least in part to the presence of phenolic compounds, and it is not toxic to renal and hepatic tissues. Novelty and scientific contribution. This bio-product can be used as an alternative to synthetic anti-inflammatory agents with the same pharmacological potential and fewer side effects. We demonstrated that Vitis labrusca winemaking waste can be used for the production of antinociceptive and anti-inflammatory products (nutraceutical, pharmaceutical and cosmetics) without toxicity, contributing to the environmental economy.Pozadina istraživanja. Ekstrakt komine grožđa, kao i vino, ima mnoga biološka svojstva, poput antioksidacijskog i protuupalnog učinka. Nažalost, proizvođači vina odbacuju neiskorišteni trop iako sadržava bioaktivne spojeve s antioksidacijskim i protuupalnim svojstvima. Svrha je ovoga rada bila ispitati antinociceptivna i protuupalna svojstva vodeno-etanolnog ekstrakta pokožice grožđa vrste Vitis labrusca iz agroindustrijskog otpada. Značaj je ovoga istraživanja u tome što se korištenjem otpada daje dodatna ekonomska vrijednost grožđu u uzgojnom lancu. Eksperimentalni pristup. Ekstrakt je dobiven iz pokožica izdvojenih iz reprezentativnog uzorka komine. Udjel fenolnih spojeva određen je praćenjem višestrukih reakcija pomoću masene spektrometrije i metodom Folin-Ciocalteu, uz galnu kiselinu kao standard. Biološka aktivnost ekstrakata ispitana je na miševima koji su hranjeni sirovim ekstraktom u dozama od 30, 100 i 300 mg/kg. Ispitani su sljedeći parametri: mehanička hiperalgezija pomoću von Frey filamenata, toplinska hiperalgezija na vrućoj ploči pri 55 °C, edem šape pomoću pomičnog mjerila i aktivnost mijeloperoksidaze kao pokazatelj aktivacije neutrofila. Nefrotoksičnost i hepatotoksičnost su ispitane biokemijskim pretragama uzoraka krvi miševa hranjenih ekstraktom grožđa vrste Vitis labrusca. Rezultati i zaključci. Maseni udjel pokožica u svim uzorcima otpada vinske industrije iznosio je 75 % mokre tvari i 59 % suhe tvari. Identificirali smo devet različitih antocijanina (3-O-glukozide peonidin, delfinidin, petunidin i malvidin; 3-p-kumaroil-glukozide cijanidin, peonidin, pe¬tunidin i malvidin, te malvidin-3,5-diglukozid), pet flavonoida (apigenin-7-glukozid, luteolin-7-glukozid, kvercetin-3-galaktozid, izorhamnetin-3-glukozid i miri¬cetin-3-rutinozid), a maseni udjel fenolnih spojeva, izražen kao ekvivalent galne kiseline, bio je 26,62 mg/g. Ispitivanja in vivo pokazala su da su ektrakti grožđa vrste Vitis labrusca masenog udjela 100 i 300 mg/kg smanjili mehaničku i toplinsku hiperalgeziju kod miševa nakon injekcije karagenana, reducirali edem šape za 50 % i smanjili broj neutrofila. Osim toga, nije bilo pokazatelja nefrotoksičnosti i hepatotoksičnosti. Ekstrakt dobiven iz otpada vinske industrije ima analgetska i protuupalna svojstva, djelomično zbog toga što sadržava fenolne spojeve, a nije toksičan za tkiva bubrega i jetre. Novina i znanstveni doprinos. Dobiveni se bioproizvod može upotrijebiti kao alternativa sintetičkim protuupalnim agensima, s istim farmakološkim potencijalom a manje nuspojava. Pokazali smo da se vinski otpad grožđa vrste Vitis labrusca može upotrijebiti za proizvodnju antinociceptivnih i protuupalnih proizvoda (nutraceutičkih, farmaceutskih i kozmetičkih) koji nemaju toksični učinak, te na taj način pridonijeti zaštiti okoliša

Budlein A, a Sesquiterpene Lactone From Viguiera robusta, Alleviates Pain and Inflammation in a Model of Acute Gout Arthritis in Mice

Background: Gout is the most common inflammatory arthritis worldwide. It is a painful inflammatory disease induced by the deposition of monosodium urate (MSU) crystals in the joints and peri-articular tissues. Sesquiterpene lactones (SLs) are secondary metabolite biosynthesized mainly by species from the family Asteraceae. It has been demonstrated that SLs present anti-inflammatory, analgesic, antitumoral, antiparasitic, and antimicrobial activities. In this study, we aimed at evaluating the efficacy of the SL budlein A in a model of acute gout arthritis in mice.Methods: Experiments were conducted in male Swiss or male LysM-eGFP mice. Animals were treated with budlein A (1 or 10 mg/kg) or vehicle 30 min before stimulus with MSU (100 μg/10 μL, intra-articular). Knee joint withdrawal threshold and edema were evaluated using electronic von Frey and caliper, respectively, 1–15 h after MSU injection. Leukocyte recruitment was determined by counting cells (Neubauer chamber), H&E staining, and using LysM-eGFP mice by confocal microscopy. Inflammasome components, Il-1β, and Tnf-α mRNA expression were determined by RT-qPCR. IL-1β and TNF-α production (in vitro) and NF-κB activation (in vitro and in vivo) were evaluated by ELISA. In vitro analysis using LPS-primed bone marrow-derived macrophages (BMDMs) was performed 5 h after stimulation with MSU crystals. For these experiments, BMDMs were either treated or pre-treated with budlein A at concentrations of 1, 3, or 10 μg/mL.Results: We demonstrated that budlein A reduced mechanical hypersensitivity and knee joint edema. Moreover, it reduced neutrophil recruitment, phagocytosis of MSU crystals by neutrophils, and Il-1β and Tnf-α mRNA expression in the knee joint. In vitro, budlein A decreased TNF-α production, which might be related to the inhibition of NF-κB activation. Furthermore, budlein A also reduced the IL-1β maturation, possibly by targeting inflammasome assembly in macrophages.Conclusion: Budlein A reduced pain and inflammation in a model of acute gout arthritis in mice. Therefore, it is likely that molecules with the ability of targeting NF-κB activation and inflammasome assembly, such as budlein A, are interesting approaches to treat gout flares

Infraspecific variation in the chemistry of glandular trichomes of two Brazilian Viguiera species (Heliantheae; Asteraceae)

A chemical survey on the glandular trichome chemistry of different populations of two Brazilian Viguiera species has been performed by the glandular trichome microsampling technique and isocratic HPLC analyses. In all six analysed samples of V. robusta, the furanoheliangolide budlein A and its tiglate and methacrylate isomers were detected as the major compounds. They were accompanied by various constituents in mostly minor or trace amounts. The chemical pattern of all samples was qualitatively very similar, thus featuring V. robusta as a taxon of high "chemoconsistency". In contrast, V. quinqueremis represented an example of "chemodiversity". Although budlein A and its derivatives were present in five of the six analysed populations, other heliangolides and germacrolides co-occurred and partly dominated in quantity. Distinct compound patterns divided the samples of V. quinqueremis into three chemical subgroups

Further sesquiterpene lactones from viguiera robusta and the potential anti-inflammatory activity of a heliangolide: Inhibition of human neutrophil elastase release

In addition to known heliangolides, a new eudesmanolide was isolated from the leaf rinse extract of Viguiera robusta (Asteraceae). Structural elucidation was based oil spectral analysis. It is the first report on eudesmanolides in members of the subgenus Calanticaria of Viguiera. In this work, the main isolated compound, the furanoheliangolide budlein A, besides its previously, reported in vitro and in vivo anti-inflammatory activities, inhibited human neutrophil elastase release. The inhibition was at the concentration of (16.83 +/- 1.96) mu M for formylated bacterial tripeptide (fMLP) stimulation and (11.84 +/- 1.62) mu M for platelet aggregation factor (PAF) stimulation, being slightly less active than the reference drug parthenolide. The results are important to demonstrate the potential anti-inflammatory activities of sesquiterpene lactones and corroborate the previous studies using other targets.CAPESFAPESPCNP

Trypanocidal Activity of Pimarane Diterpenes from Viguiera arenaria (Asteraceae)

Five structurally related pimarane diterpenes isolated from the roots of Viguiera arenaria and a further compound obtained by chemical derivatization were evaluated in vitro against the trypomastigote forms of Trypanosoma cruzi. The natural compound ent-15-pimarene-8 beta,19-diol and the derivative ent-8(14),15-pimaradiene-3 beta-acetoxy showed the highest trypanocidal activity, displaying IC50 values of 116.5 +/- 1.21 and 149.3 +/- 1.07 mu M, respectively, while the positive control, violet gentian, showed an IC50 of 76 mu M. Based on the results, it can be concluded that minor structural differences among the tested diterpenes influence significantly the trypanocidal activity, thus bringing new perspectives to the establishment of structure-activity relationships among this type of metabolites to the treatment of Chagas` disease. Copyright (C) 2008 John Wiley & Sons, Ltd.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq

Budlein A from Viguiera robusta inhibits leukocyte-endothelial cell interactions, adhesion molecule expression and inflammatory mediators release

Budlein A has been reported to exert some analgesic and anti-inflammatory properties. In this study, we have evaluated its effect on LPS-induced leukocyte recruitment in vivo and the mechanisms involved in its anti-inflammatory activity. In vivo, intravital videomicroscopy was used to determine the effects of budlein A on LPS-induced leukocyte-endothelial cell interactions in the murine cremasteric microcirculation. In vitro, the effects of budlein A on LPS-induced cytokine, chemokine and nitrites release, T-cell proliferative response as well as cell adhesion molecule expression (CAM) were evaluated. In vivo, intraperitoneal administration of budlein A (2.6 mM/kg) caused a significant reduction of LPS-induced leukocyte rolling flux, adhesion and emigration by 84, 92 and 96% respectively. In vitro, T-cell proliferative response was also affected by budlein A. When murine J774 macrophages were incubated with the sesquiterpene lactone, LPS-induced IL-1 beta, tumor necrosis factor-alpha (TNF-alpha) and keratinocyte-derived chemokine (KC) release were concentration-dependently inhibited. In human umbilical vein endothelial cells (HUVECs), budlein A also reduced the production of TNF-alpha, monocyte chemoattractant protein-1 (MCP-1), IL-8, nitrites and CAM expression elicited by LPS. Budlein A is a potent inhibitor of LPS-induced leukocyte accumulation in vivo. This effect appears to be mediated through inhibition of cytokine and chemokine release and down-regulation of CAM expression. Thus, it has potential therapeutic interest for the control of leukocyte recruitment that occurs in different inflammatory disorders. (C) 2009 Elsevier GrnbH. All rights reserved.CICYT[SAF 2008-03477]Conselleria of Education and Culture (Generalitat Valenciana)[03/166]Funda do de Amparo A Pesquisa do Estado de Sao Paulo (FAPESP) (Brazil)Programa de Mobilidade IniernacionalSpanish Ministry of Education and ScienceFAPESPUniversity of Valencia, Spai

Kaurenoic Acid from Sphagneticola trilobata Inhibits Inflammatory Pain: Effect on Cytokine Production and Activation of the NO-Cyclic GMP-Protein Kinase G-ATP-Sensitive Potassium Channel Signaling Pathway

Kaurenoic acid [ent-kaur-16-en-19-oic acid (1)] is a diterpene present in several plants including Sphagneticola trilobata. The only documented evidence for its antinociceptive effect is that it inhibits the writhing response induced by acetic acid in mice. Therefore, the analgesic effect of 1 in different models of pain and its mechanisms in mice were investigated further. Intraperitoneal and oral treatment with 1 dose-dependently inhibited inflammatory nociception induced by acetic acid. Oral treatment with 1 also inhibited overt nociception-like behavior induced by phenyl-p-benzoquinone, complete Freund's adjuvant (CFA), and both phases of the formalin test. Compound 1 also inhibited acute carrageenin- and PGE(2)-induced and chronic CFA-induced inflammatory mechanical hyperalgesia. Mechanistically, 1 inhibited the production of the hyperalgesic cytokines TNF-alpha and IL-1 beta. Furthermore, the analgesic effect of 1 was inhibited by L-NAME, ODQ, KT5823, and glybenclamide treatment, demonstrating that such activity also depends on activation of the NO-cyclic GMP-protein kinase G-ATP-sensitive potassium channel signaling pathway, respectively. These results demonstrate that 1 exhibits an analgesic effect in a consistent manner and that its mechanisms involve the inhibition of cytokine production and activation of the NO-cyclic GMP-protein lcinase G-ATP-sensitive potassium channel signaling pathway.Fundacao de Amparo a Pesquisa do Estado de So Paulo (FAPESP, Brazil)Fundacao de Amparo a Pesquisa do Estado de So Paulo (FAPESP, Brazil)Conselho Nacional de Pesquisa (CNPq, Brazil)Conselho Nacional de Pesquisa (CNPq, Brazil)Coordenadoria de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), BrazilCoordenadoria de Aperfeicoamento de Pessoal de Nivel Superior (CAPES, Brazil)Fundacao Araucaria and Governo do Estado do Parana (Brazil)Fundacao Araucaria and Governo do Estado do Parana (Brazil