RESUMEN
Cell penetrating peptides (CPPs) are usually short, highly cationic peptides that are capable of crossing the cell membrane and transport cargos of varied size and nature in cells by energy- and receptor-independent mechanisms. An additional potential is the newly discovered anti-tumor activity of certain CPPs, including RW16 (RRWRRWWRRWWRRWRR) which is derived from penetratin and is investigated here. The use of CPPs in therapeutics, diagnosis and potential application as anti-tumor agents increases the necessity of understanding their mode of action, a subject yet not totally understood. With this in mind, the membrane interaction and perturbation mechanisms of RW16 with both zwitterionic and anionic lipid model systems (used as representative models of healthy vs tumor cells) were investigated using a large panoply of biophysical techniques. It was shown that RW16 autoassociates and that its oligomerization state highly influences its membrane interaction. Overall a stronger association and perturbation of anionic membranes was observed, especially in the presence of oligomeric peptide, when compared to zwitterionic ones. This might explain, at least in part, the anti-tumor activity and so the selective interaction with cancer cells whose membranes have been shown to be especially anionic. Hydrophobic contacts between the peptide and lipids were also shown to play an important role in the interaction. That probably results from the tryptophan insertion into the fatty acid lipid area following a peptide flip after the first electrostatic recognition. A model is presented that reflects the ensemble of results.
Asunto(s)
Antibacterianos/metabolismo , Antineoplásicos/metabolismo , Permeabilidad de la Membrana Celular , Membrana Celular/metabolismo , Péptidos de Penetración Celular/metabolismo , Lípidos de la Membrana/metabolismo , Antibacterianos/química , Antibacterianos/farmacología , Antineoplásicos/química , Antineoplásicos/farmacología , Péptidos de Penetración Celular/química , Péptidos de Penetración Celular/farmacología , Escherichia coli/efectos de los fármacos , Escherichia coli/crecimiento & desarrollo , Interacciones Hidrofóbicas e Hidrofílicas , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/crecimiento & desarrollo , Liposomas , Pruebas de Sensibilidad Microbiana , Conformación Proteica , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/crecimiento & desarrollo , TriptófanoRESUMEN
BACKGROUND: Viticultural residues from commercial viticultural activities represent a potentially important source of bioactive stilbenes such as resveratrol. The main aim of the present study was therefore to isolate, identify and perform biological assays against amyloid-ß peptide aggregation of original stilbenes from Vitis vinifera shoots. RESULTS: A new resveratrol oligomer, (Z)-cis-miyabenol C (3), was isolated from Vitis vinifera grapevine shoots together with two newly reported oligostilbenes from Vitis vinifera shoots, vitisinol C (1) and (E)-cis-miyabenol C (2), and six known compounds: piceatannol, resveratrol, (E)-ε-viniferin (trans-ε-viniferin), ω-viniferin, vitisinol C and (E)-miyabenol C. The structures of these resveratrol derivatives were established on the basis of detailed spectroscopic analysis including nuclear magnetic resonance experiments. All the newly reported compounds were tested for their anti-aggregative activity against amyloid-ß fibril formation. Vitisinol C was found to exert a significant activity against amyloid-ß aggregation. CONCLUSION: Vitis vinifera grapevine shoots are potentially interesting as a source of new bioactive stilbenes, such as vitisinol C.
Asunto(s)
Descubrimiento de Drogas , Residuos Industriales/análisis , Nootrópicos/aislamiento & purificación , Extractos Vegetales/química , Brotes de la Planta/química , Estilbenos/aislamiento & purificación , Vitis/química , Agricultura/economía , Péptidos beta-Amiloides/antagonistas & inhibidores , Péptidos beta-Amiloides/metabolismo , Benzofuranos/análisis , Benzofuranos/química , Benzofuranos/economía , Benzofuranos/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Francia , Humanos , Residuos Industriales/economía , Estructura Molecular , Fármacos Neuroprotectores/química , Fármacos Neuroprotectores/economía , Fármacos Neuroprotectores/aislamiento & purificación , Fármacos Neuroprotectores/farmacología , Nootrópicos/química , Nootrópicos/economía , Nootrópicos/farmacología , Fragmentos de Péptidos/antagonistas & inhibidores , Fragmentos de Péptidos/metabolismo , Fenoles/química , Fenoles/economía , Extractos Vegetales/economía , Agregado de Proteínas/efectos de los fármacos , Agregación Patológica de Proteínas , Espectrometría de Masa por Ionización de Electrospray , Estereoisomerismo , Estilbenos/análisis , Estilbenos/química , Estilbenos/economía , Estilbenos/farmacología , EstilbestrolesRESUMEN
The aggregation of α-synuclein is one on the key pathogenic events in Parkinson's disease. In the present study, we investigated the inhibitory capacities of stilbenes against α-synuclein aggregation and toxicity. Thioflavin T fluorescence, transmission electronic microscopy, and SDS-PAGE analysis were performed to investigate the inhibitory effects of three stilbenes against α-synuclein aggregation: piceatannol, ampelopsin A, and isohopeaphenol. Lipid vesicle permeabilization assays were performed to screen stilbenes for protection against membrane damage induced by aggregated α-synuclein. The viability of PC12 cells was examined using an MTT assay to assess the preventive effects of stilbenes against α-synuclein-induced toxicity. Piceatannol inhibited the formation of α synuclein fibrils and was able to destabilize preformed filaments. It seems to induce the formation of small soluble complexes protecting membranes against α-synuclein-induced damage. Finally, piceatannol protected cells against α-synuclein-induced toxicity. The oligomers tested (ampelopsin A and hopeaphenol) were less active.
Asunto(s)
Estilbenos/farmacología , Vino/análisis , alfa-Sinucleína/toxicidad , Animales , Benzotiazoles , Supervivencia Celular/efectos de los fármacos , Electroforesis en Gel de Poliacrilamida , Flavonoides/farmacología , Microscopía Electrónica de Transmisión , Células PC12 , Enfermedad de Parkinson/tratamiento farmacológico , Fenoles , Ratas , Tiazoles , alfa-Sinucleína/antagonistas & inhibidoresRESUMEN
Several cultivars of peach fruit (Prunus persica L.) were investigated. Their phenolic composition and concentration were assessed by LC-MS. Concentrations were calculated in mg per g of dry weight extract. Their antioxidant capacity (Folin-Ciocalteu, ORAC, DPPH, ABTS, PFRAP and ICA), inhibitory property against ß-amyloid and α-synuclein fibril formation and protective capacity against Aß-induced toxicity on PC12 cell lines (viability assessed by MTT assay and intracellular ROS production by DCFH-DA assay) were evaluated. Fifteen different phenolic compounds were identified and quantified. In particular, new isorhamnetin derivatives were identified. Phenolic contents were ranged between 19 and 82mg/g. Spring Belle extract had the highest content and Romea the lowest. Except for the ICA assay, a good correlation between phenolic content and the antioxidant capacities of peach fruit extracts was found, indicating that phenolic compounds are major contributors to their antioxidant capacity. Results indicate that the phenolic extract of peach cultivars inhibits Aß and αS fibril formation and protects PC12 cell lines against Aß-induced toxicity.
Asunto(s)
Frutas/química , Fenoles/análisis , Prunus persica , Animales , Antioxidantes/análisis , Antioxidantes/farmacología , Cromatografía Liquida , Fluoresceínas , Espectrometría de Masas , Células PC12 , Fenoles/química , Fenoles/farmacología , Ratas , Especies Reactivas de Oxígeno/químicaRESUMEN
The flavonol profiles of grape berry skins were analysed in order to assess phenotypic variation between six grapevines belonging to six different species: Vitis vinifera, Vitiscandicans, Vitischampinii, Vitisamurensis, Vitiscinerea and Vitisdoaniana. High-performance liquid chromatography coupled to mass spectrometry (LC-MS) and NMR spectrometry (LC-NMR) were used to separate and identify the flavonols present in these species. The combination of LC-MS and LC-NMR data resulted in the identification of eighteen flavonols. In particular, the new flavonol diglycoside and pentoside derivatives were determined. In addition, the antioxidant capacities of flavonol grape skin extracts were evaluated by using an oxygen radical absorbance capacity method (ORAC).