RESUMO
Silver linden (Tilia tomentosa Moench, TtM) flowers possess several health-promoting properties, especially at the neurological level, such as intestinal relaxation activity associated with specific flavonols, particularly quercetin and kaempferol derivatives. However, such molecules are susceptible to degradation upon different triggers like heat, light and extreme pH values. To overcome the scarce stability of TtM flowers bioactive molecules and make them suitable for developing functional food and supplements, we applied microencapsulation. Spray-drying microencapsulation of TtM flowers extract was performed using three starch-derived wall materials: maltodextrin 12 DE (MD12) and 19 DE (MD19), and OSA-modified starch (OSA-S). The stability of total phenols, flavanols, and antioxidant capacity was monitored for 70 days under accelerated stress conditions (40 °C/70% RH) by HPLC and spectrophotometric methods, and the intestinal contractile activity was tested in a murine model. In comparison to MD12 and MD19, OSA-S stood out for the higher encapsulation efficiency of quercetin and kaempferol glycosides (+ 36-47% compared to MD12 and + 18-24% compared to MD19) and stability thereof (half-life on average + 30% compared to MD12 and + 51% compared to MD19). The intestinal contractile activity of OAS-S powders resulted comparable to the original extract, indicating that flavonols were biologically active and accessible. Our results underly the potential advantages of OSA-S encapsulated formulation as a functional ingredient for the development of nutraceutical products.
Assuntos
Tilia , Animais , Camundongos , Flores/química , Quempferóis/análise , Extratos Vegetais/química , Quercetina/análise , Amido/química , Tilia/químicaRESUMO
Microalgae and microalgae-derived compounds have great potential as supplements in the human diet and as a source of bioactive products with health benefits. Spirulina (Arthrospira platensis (Nordstedt) Gomont, or Spirulina platensis) belongs to the class of cyanobacteria and has been studied for its numerous health benefits, which include anti-inflammatory properties, among others. This work was aimed at comparing some spirulina products available on the Italian market. The commercial products here analyzed consisted of spirulina cultivated and processed with different approaches. Single-component spirulina products in powder and flake form, free of any type of excipient produced from four different companies operating in the sector, have been analyzed. The macro- and micromorphological examination, and the content of pigments, phycobiliproteins, phenols, and proteins have shown differences regarding the morphology and chemical composition, especially for those classes of particularly unstable compounds such as chlorophylls and carotenoids, suggesting a great influence of both culture conditions and processing methods.
Assuntos
Microalgas , Spirulina , Animais , Carotenoides/química , Decapodiformes , Humanos , Pós , Spirulina/químicaRESUMO
Dietary supplementation with nutrients able to control intestinal and systemic inflammation is of marketable interest. Indeed, gastrointestinal homeostasis plays a significant role in maintaining human health. In this setting, E. gracilis may sustain or promote human health, but the effects on the intestinal inflammatory milieu are not clear. In this study, we investigated the anti-inflammatory activity of E. gracilis and inferred possible mechanisms. Paramylon, crude, and fractionated extracts were obtained from E. gracilis grown in vitro. Phytoconstituents of the extracts were characterized using TLC and HPLC UV-Vis. The anti-inflammatory and antioxidant activities were investigated in primary human macrophages and an intestinal epithelial cell line (HT-29). The analysis of the extracts led to identifying ß-carotene, neoxanthin, diadinoxanthin, canthaxanthin, and breakdown products such as pheophytins and pheophorbides. E. gracilis fractionated extracts reduced the production of tumor necrosis factor-α triggered by bacterial lipopolysaccharide (LPS) in the short and long terms. Pheophytin a and b and canthaxanthin increased the intracellular reducing potential and dampened the production of LPS-induced reactive oxygen species and lipid peroxidation, intracellular events usually involved in the perpetuation of chronic inflammatory disorders. This study rationalizes the role of specific extract fractions of E. gracilis in controlling LPS-driven intestinal inflammation.
RESUMO
Functional gastrointestinal disorders (FGIDs) are characterized by abdominal pain, bloating and bowel disturbances. FGID therapy is primarily symptomatic, including treatment with herbal remedies. Flower extract of Tilia tomentosa Moench (TtM) is occasionally used as an anti-spasmodic in popular medicine. Since its effect on intestinal response is unknown, we evaluated the influence of TtM extract on small intestine contractility. Ileal preparations from C57BL/6J mice were mounted in organ baths to assess changes in muscle tension, following addition of TtM extract (0.5-36 µg/mL) or a vehicle (ethanol). Changes in contractile response to receptor- and non-receptor-mediated stimuli were assessed in ileal preparations pretreated with 12 µg/mL TtM. Alterations in the enteric nervous system neuroglial network were analyzed by confocal immunofluorescence. Increasing addition of TtM induced a marked relaxation in ileal specimens compared to the vehicle. Pretreatment with TtM affected cholinergic and tachykininergic neuromuscular contractions as well as K+-induced smooth muscle depolarization. Following incubation with TtM, a significant reduction in non-adrenergic non-cholinergic-mediated relaxation sensitive to Nω-Nitro-L-arginine methyl ester hydrochloride (pan-nitric oxide synthase inhibitor) was found. In vitro incubation of intestinal specimens with TtM did not affect the myenteric plexus neuroglial network. Our findings show that TtM-induced intestinal relaxation is mediated by nitric oxide pathways, providing a pharmacological basis for the use of TtM in FGIDs.
Assuntos
Intestino Delgado/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Extratos Vegetais/farmacologia , Tilia , Animais , Gastroenteropatias/tratamento farmacológico , Gastroenteropatias/fisiopatologia , Íleo/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BLRESUMO
Melaleuca alternifolia tea tree oil (TTO) is largely used in cutaneous infections. Clinical observations reported antibacterial, antifungal, and antiviral activities, whereas in vitro experiments ascribed most of biological properties to terpinen-4-ol. Since different plant chemotypes and storage conditions result in variations of chemical composition of commercially available TTO, in this study we investigated the antimicrobial activity and the chemical profile of ten commercially available TTO products. The antimicrobial activity was assessed against Candida glabrata, Herpes simplex virus type 1 (HSV-1), methicillin-resistant Staphylococcus aureus (MRSA), and Pseudomonas aeruginosa grown in planktonic mode or biofilms. Only five out of ten TTO batches reported significant antimicrobial activity. The identified TTO products reduced bacterial survival in biofilms, generated oxidative damage in C. glabrata, and diminished HSV-1 infectivity. GC-MS analysis revealed that all the analyzed TTO batches fitted into the terpinen-4-ol chemotype even if we reported great variability in composition of nine major ISO-specified TTO components. Overall, we were not able to ascribe the antimicrobial activity to the content in terpinen-4-ol. We therefore conclude that the antimicrobial activity of TTO results from complex interaction among different components.
Assuntos
Anti-Infecciosos/farmacologia , Candida glabrata/crescimento & desenvolvimento , Herpesvirus Humano 1/crescimento & desenvolvimento , Melaleuca/química , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Pseudomonas aeruginosa/crescimento & desenvolvimento , Óleo de Melaleuca/farmacologia , Biofilmes/efeitos dos fármacos , Candida glabrata/efeitos dos fármacos , Herpesvirus Humano 1/efeitos dos fármacos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Terpenos/farmacologiaRESUMO
The antiproliferative effect of the naturally occurring polyphenol chlorogenic acid (CGA) was evaluated in combination with either cisplatin or oxaliplatin in human cervical carcinoma cell lines that were either sensitive (A431) or resistant to cisplatin (A431Pt), in order to provide evidence to overcome drug resistance. Cytotoxicity of platinating drugs (IC50 - 10(-6) - 10(-5) M) was enhanced by 1-2 orders of magnitude by increasing incubation times (1, 4, and 24 hours) in the two cell lines. CGA treatment presented low cytotoxicity per se (IC50 ~ 10(-4) M at 24 h) if compared with platinum drugs and its activity was similar in A431Pt cells and in their sensitive A431 counterpart. The combination of the platinating drugs with CGA (10(-6) - 10(-4) M) indicated variable effects on cytotoxicity, ranging from potentiation to various degrees of antagonism (in A431 cells) and no effect (in A431Pt cells). In order to explain the different cytotoxic activity elicited by oxaliplatin and cisplatin in association with CGA, the possible presence of chemical interactions was investigated by HPLC analysis. The drug association with CGA caused evident changes in their chromatographic profile, suggesting occurrence of in vitro chemical interactions.
Assuntos
Antineoplásicos/uso terapêutico , Ácido Clorogênico/uso terapêutico , Cisplatino/uso terapêutico , Compostos Organoplatínicos/uso terapêutico , Neoplasias do Colo do Útero/tratamento farmacológico , Linhagem Celular Tumoral , Sinergismo Farmacológico , Feminino , Humanos , OxaliplatinaRESUMO
Studies have evidenced the biological properties of fucoxanthin and the commercialisation of various nutritional supplements based on seaweed extracts that claim high fucoxanthin content is constantly growing. However, it was demonstrated that fucoxanthin is highly susceptible to degradation, and in this context the need for simple and reliable analytical methods for fucoxanthin analysis is evident. This article presents a simple thin-layer chromatography (TLC) densitometric method for the determination of fucoxanthin from Undaria pinnatifida extracts and commercial formulations. Fucoxanthin was separated on TLC silica gel 60 F254 plates with acetone:n-hexane 6:4 v/v as mobile phase. Densitometric analysis was performed using the Image J image-processing program available online. Calibration data revealed good linear relationship (R(2) = 0.9955) between peak area and concentration of fucoxanthin. The repeatability was 2.0 (% CV), and intra-day and inter-day precisions were 1.5 and 2.0 (% RSD), respectively. The proposed method provides an inexpensive tool in preliminary evaluation of fucoxanthin-based products.
Assuntos
Cromatografia em Camada Fina/métodos , Alga Marinha/química , Xantofilas/análise , Calibragem , Densitometria/métodos , HexanosRESUMO
Coronilla scorpioides (L.) W.D.J. Koch is known for producing several compounds with pharmaceutical interest, such as the hydroxycoumarins umbelliferone, scopoletin and daphnoretin, the dihydrofuranocoumarin marmesin, and the furocoumarin psoralen. In vitro callus cultures of C. scorpioides were established from hypocotyl, leaf, stem internode and root explants in order to evaluate the possibility of in vitro production of these active secondary metabolites. Calli were obtained with high frequency from all the explant types both in B5 and MS medium. However, after the third subculture, B5 medium, giving the best results, was selected for subsequent transfers. Homogeneous calli were kept either in darkness or in light. Chemical analyses showed that scopoletin and the intermediate products of the biogenetic pathway of psoralen, umbelliferone and marmesin, were always present in the calli and excreted into the media, while daphnoretin was never detected. Light seems to be a prerequisite for psoralen biosynthesis. Root-derived calli produced a significantly higher amount of psoralen (137.5 microg g(-1) DW). Principal component analysis showed that umbelliferone, marmesin and psoralen contents are related to variables associated with different explant types.
Assuntos
Cumarínicos/metabolismo , Fabaceae/metabolismo , Cumarínicos/química , Fabaceae/química , Germinação , Folhas de Planta/citologia , Raízes de Plantas/citologia , Caules de Planta/citologia , Sementes/fisiologia , Técnicas de Cultura de TecidosRESUMO
Fucoxanthin is one of the most abundant carotenoids and possesses a number of beneficial medicinal qualities which include its anti-oxidant, anti-obesity and anti-cancer properties. In this study, the photostability of fucoxanthin in extracts with different chemical profiles was studied. The extracts were obtained from Undaria pinnatifida, a seaweed rich in this carotenoid, using conventional liquid solvent extraction procedures and the QuEChERS method. All the extracts contained all-trans-fucoxanthin as the major compound. Conventional procedures produced a fucoxanthin purity of lower than 50%, whereas after liquid-liquid partition, PSA cleanup, and PSA and GCB cleanup (QuEChERS method) fucoxanthin purity increased to 70%, 86%, and 94%, respectively. Although in the acetone extract the initial content of fucoxanthin was the highest, results demonstrate that coextractives play an important role in enhancing the rate of photodegradation. After light exposure, the conventional extracts lost around 90% of the initial fucoxanthin content. On the other hand, the extracts obtained by the QuEChERS method showed significantly higher light stability than the conventional extracts. These results suggest that the QuEChERS method could be used and further improved to obtain more purified and stable extracts for fucoxanthin from U. pinnatifida.
Assuntos
Undaria/química , Xantofilas/química , Cromatografia Líquida de Alta Pressão , Extratos Vegetais/análise , Extratos Vegetais/química , Xantofilas/análiseRESUMO
The Crambe tataria glucosinolate/myrosinase system in seeds and leaves of in vivo and in vitro regenerated plantlets, and two callus cell lines was investigated. It was demonstrated that in all the extracts glucosinolates were present and the myrosinase system was operative. There appears to be no discrimination between the glucosinolates used as substrates, but the hydrolysis rates were different regardless of the nature of the side chain. This is one of the first studies demonstrating that undifferentiated cells are able to synthetize glucosinolates and have an operating myrosinase system.
Assuntos
Crambe (Planta)/metabolismo , Glucosinolatos/biossíntese , Glicosídeo Hidrolases/fisiologia , Cromatografia Líquida de Alta PressãoRESUMO
A method based on FIA-ESI-MS has been developed to profile the major constituents of Hypericum perforatum extracts. The objective was to obtain simultaneous semi-quantitative data on hypericin, chlorogenic acid, rutin, quercitrin, quercetin and hyperforin contents for a high-throughput screening of the raw plant material. The principal drawback of FIA-ESI analysis of complex mixtures involves ion suppression effects: the ionization of some components of the mixture can be severely suppressed by ionization of others. The results show that this problem can be alleviated using a new approach to generate calibration curves.
Assuntos
Hypericum/química , Calibragem , Cromatografia Líquida de Alta Pressão , Indicadores e Reagentes , Extratos Vegetais/química , Padrões de Referência , Espectrometria de Massas por Ionização por Electrospray , Espectrofotometria UltravioletaRESUMO
St. John's Wort extracts are used for the treatment of mild to moderately severe depression, and their composition and standardization have been thoroughly investigated. Standardization of St. John's Wort extracts has to cope with several factors that affect the phytochemical profile of the plant, with geographic location, seasonal variations and subspecies all being important. To address this issue, three different subspecies of Hypericum perforatum L. were profiled in relation to different maturation phases, evaluating the variations in the context of the major secondary metabolites from this plant. HPLC analysis indicated that H. perforatum subsp. perforatum is richer in secondary metabolites than the other subspecies, and that the three subspecies show a different profile during the developmental stages. Hypericins, hyperforins and flavonoids peak at quite different stages, and the standardization of Hyperici extracts based only on the contents of hypericin is not adequate to guarantee batch reproducibility of the extracts.
Assuntos
Antidepressivos/metabolismo , Flavonoides/metabolismo , Hypericum/metabolismo , Perileno/análogos & derivados , Floroglucinol/análogos & derivados , Extratos Vegetais/química , Terpenos/metabolismo , Antracenos , Compostos Bicíclicos com Pontes/metabolismo , Flavonoides/análise , Flores , Hypericum/química , Hypericum/genética , Perileno/metabolismo , Floroglucinol/metabolismo , Extratos Vegetais/normas , Folhas de Planta , Padrões de Referência , Especificidade da EspécieRESUMO
The biologically active naphthodianthrones hypericin and pseudohypericin were detected by electrospray ionization mass spectrometry (ESI-MS/MS) in microsamples from the sepals of Hypericum elodes (Hypericaceae) containing the so-called "red glands", i.e. stipitate glands with red-coloured heads. The occurrence of hypericins in the red glands of H. elodes supports the taxonomic position of the section Elodes within the genus Hypericum and provides evidence that the ability of carrying out the biosynthetic pathway leading to the naphthodianthrone compounds, rather than the absolute amounts produced, should be regarded as a chemical marker of the phylogenetically more advanced sections of genus Hypericum. The biologically active phloroglucinol derivatives hyperforin and adhyperforin, so far found only in H. perforatum, were also detected and evidence for their localization in the sepal secretory canals with large lumen, is given.
Assuntos
Hypericum/química , Perileno/análogos & derivados , Perileno/isolamento & purificação , Antracenos , Flores/química , Microscopia Eletrônica de Varredura , Perileno/química , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
A widespread occurrence of melatonin in plant kingdom has been reported. The circadian rhythm in the level of melatonin observed in both unicellular algae and higher plants, suggests a role in regulation of photoperiodic and rhythmic phenomena, i.e. a similar function for melatonin in both plants and animals. Evidence has been obtained for a role of melatonin in plant morphogenesis, but more research is needed to ascertain other suggested physiological roles in higher plants (seed dormancy regulation, radical scavenger activity, interaction with calmodulin) as well the ecological significance of the high melatonin levels recorded in alpine plants. Setting-up more reliable analytical methods for melatonin detection and quantification is a basic requirement to get more insight into melatonin roles in plant physiology and ecology.