RESUMEN
Historically, natural products have been the most successful source of inspiration for the development of new drugs. Members of the Thymelaeaceae family have been of interest owing to their excellent medicinal value. Given the successful history of natural product-based drug discovery, extracts from the aerial parts of Thymelaea hirsuta were essvaluated for their potential anti-human immunodeficiency virus type 1 (HIV-1) activity. Ethyl acetate extracts from leaves (71B) and branches (72B) of Thymelaea hirsuta showed potent and selective activity against HIV-1 wt (EC50 = 0.8 µg/mL) at non-cytotoxic concentrations (CC50 > 100 µg/mL). They proved to be active against HIV-1 variants carrying clinically relevant NNRTI and NRTI mutations at low concentration (0.3-4 µg/mL range) and against the M-tropic strain HIV-1 BaL. The 72B extract, chosen as a lead, was not able to inhibit the RT and protease enzymatic functions. Furthermore, it was not virucidal, since exposure of HIV to high concentration did not affect virus infectivity. The pre-clinical safety profile of this extract showed no adverse effect on the growth of Lactobacilli, and non-toxic concentration of the extract did not influence the Caco-2 epithelial cells monolayer integrity. Additionally, extract 72B prevented syncytia formation at low concentration (0.4 µg/mL). The potent inhibitory effect on the syncytia formation in co-cultures showed that 72B inhibits an early event in the replication cycle of HIV. All of these findings prompt us to carry on new studies on Thymelaea hirsuta extracts.
Asunto(s)
Fármacos Anti-VIH/farmacología , Descubrimiento de Drogas/métodos , Infecciones por VIH/tratamiento farmacológico , Extractos Vegetales/farmacología , Thymelaeaceae/química , Animales , Células CACO-2 , Bovinos , Línea Celular Tumoral , Chlorocebus aethiops , Cricetinae , VIH-1/efectos de los fármacos , Humanos , Hojas de la Planta/química , Células VeroRESUMEN
OBJECTIVE: To investigate the link between Human Herpesvirus 8 (HHV8) infection and plasma oxidative stress in patients with diabetes mellitus type 2 (DM2). RESULTS: Blood samples collected from DM2 and control subjects were screened for the presence of antibodies against HHV8 and for biomarkers of oxidative stress. We determined the products of radical damage on the plasma lipid fraction, such as malondialdehyde (MDA), fatty acid hydroperoxides (HP) and 7-ketocholesterol (7-keto), the oxidation products of unsaturated fatty acids (UFA) and cholesterol, respectively. The level of plasma antioxidant α-tocopherol (α-toc) was also assessed. Relevant differences were observed in the redox status in DM2 and either HHV8-positive or -negative control subjects. The level of α-toc significantly decreased in both DM2 and HHV8-positive subjects. Levels of MDA, HP and 7-keto were much higher in HHV8-positive and DM2 subjects, indicating that plasma oxidative stress is a common feature in both DM2 and HHV8-infection. In addition, 7-keto was further increased in HHV8-positive DM2 patients. We hypothesized that the HHV8-infection may contribute to the production of ROS, and hence to the oxidative stress closely related to the pathogenesis and development of DM2.
Asunto(s)
Diabetes Mellitus Tipo 2/patología , Diabetes Mellitus Tipo 2/virología , Infecciones por Herpesviridae/complicaciones , Herpesvirus Humano 8/fisiología , Estrés Oxidativo , Adulto , Anciano , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2/sangre , Femenino , Infecciones por Herpesviridae/sangre , Humanos , Cetocolesteroles/sangre , Masculino , Malondialdehído/sangre , Persona de Mediana Edad , alfa-Tocoferol/sangreRESUMEN
Dietary habits may strongly influence intestinal homeostasis. Oxysterols, the oxidized products of cholesterol present in cholesterol-containing foodstuffs, have been shown to exert pro-oxidant and pro-inflammatory effects, altering intestinal epithelial layer and thus contributing to the pathogenesis of human inflammatory bowel diseases and colon cancer. Extra virgin olive oil polyphenols possess antioxidant and anti-inflammatory properties, and concentrate in the intestinal lumen, where may help in preventing intestinal diseases. In the present study we evaluated the ability of an extra virgin olive oil phenolic extract to counteract the pro-oxidant and pro-inflammatory action of a representative mixture of dietary oxysterols in the human colon adenocarcinoma cell line (Caco-2) undergoing full differentiation into enterocyte-like cells. Oxysterols treatment significantly altered differentiated Caco-2 cells redox status, leading to oxidant species production and a decrease of GSH levels, after 1â¯h exposure, followed by an increase of cytokines production, IL-6 and IL-8, after 24â¯h. Oxysterol cell treatment also induced after 48â¯h an increase of NO release, due to the induction of iNOS. Pretreatment with the phenolic extract counteracted oxysterols effects, at least in part by modulating one of the main pathways activated in the cellular response to the action of oxysterols, the MAPK-NF-kB pathway. We demonstrated the ability of the phenolic extract to directly modulate p38 and JNK1/2 phosphorylation and activation of NF-kB, following its inhibitor IkB phosphorylation. The phenolic extract also inhibited iNOS induction, keeping NO concentration at the control level. Our results suggest a protective effect at intestinal level of extra virgin olive oil polyphenols, able to prevent or limit redox unbalance and the onset and progression of chronic intestinal inflammation.
Asunto(s)
Antioxidantes/farmacología , Inflamación/prevención & control , Aceite de Oliva/farmacología , Polifenoles/farmacología , Células CACO-2 , Humanos , Inflamación/genética , Inflamación/metabolismo , Inflamación/patología , Interleucina-6/biosíntesis , Interleucina-8/biosíntesis , Mucosa Intestinal/metabolismo , Intestinos/efectos de los fármacos , FN-kappa B/genética , Óxido Nítrico/biosíntesis , Oxidación-Reducción/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Oxiesteroles/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
The emerging role of the diet in the incidence of intestinal inflammatory diseases has stimulated research on the influence of eating habits with pro-inflammatory properties in inducing epithelial barrier disturbance. Cholesterol oxidation products, namely oxysterols, have been shown to promote and sustain oxidative/inflammatory reactions in human digestive tract. This work investigated in an in vitro model the potential ability of a combination of dietary oxysterols representative of a hyper-cholesterol diet to induce the loss of intestinal epithelial layer integrity. The components of the experimental mixture were the main oxysterols stemming from heat-induced cholesterol auto-oxidation, namely 7-ketocholesterol, 5α,6α-and 5ß,6ß-epoxycholesterol, 7α- and 7ß-hydroxycholesterol. These compounds added to monolayers of differentiated CaCo-2 cells in combination or singularly, caused a time-dependent induction of matrix metalloproteinases (MMP)-2 and -9, also known as gelatinases. The hyperactivation of MMP-2 and -9 was found to be associated with decreased levels of the tight junctions zonula occludens-1 (ZO-1), occludin and Junction Adhesion Molecule-A (JAM-A). Together with such a protein loss, particularly evident for ZO-1, a net perturbation of spatial localization of the three tight junctions was observed. Cell monolayer pre-treatment with the selective inhibitor of MMPs ARP100 or polyphenol (-)-epicathechin, previously shown to inhibit NADPH oxidase in the same model system, demonstrated that the decrease of the three tight junction proteins was mainly a consequence of MMPs induction, which was in turn dependent on the pro-oxidant property of the oxysterols investigated. Although further investigation on oxysterols intestinal layer damage mechanism is to be carried on, the consequent - but incomplete - prevention of oxysterols-dependent TJs alteration due to MMPs inhibition, avoided the loss of scaffold protein ZO-1, with possible significant recovery of intestinal monolayer integrity.
Asunto(s)
Colesterol/análogos & derivados , Hidroxicolesteroles/farmacología , Cetocolesteroles/farmacología , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Uniones Estrechas/efectos de los fármacos , Células CACO-2 , Catequina/farmacología , Moléculas de Adhesión Celular/genética , Moléculas de Adhesión Celular/metabolismo , Colesterol/farmacología , Colesterol en la Dieta/metabolismo , Colesterol en la Dieta/farmacología , Impedancia Eléctrica , Activación Enzimática/efectos de los fármacos , Regulación de la Expresión Génica , Humanos , Peroxidación de Lípido , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Ocludina/genética , Ocludina/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Uniones Estrechas/metabolismo , Uniones Estrechas/ultraestructura , Proteína de la Zonula Occludens-1/genética , Proteína de la Zonula Occludens-1/metabolismoRESUMEN
The phenolic fraction of a naturally fermented cultivar of table olives, "Tonda di Cagliari," was investigated for the ability to protect Caco-2 cells against oxidative stress and membrane damage induced by tert-butyl hydroperoxyde (TBH). TBH exposure resulted in an alteration of cellular redox status, with an increase in reactive oxygen species (ROS) and a decrease in reduced glutathione (GSH) level. A loss of the epithelial integrity, as indicated by the decrease of the transepithelial electrical resistance value, was also observed over time, together with an intense lipid peroxidation process. The olives phenolic extract significantly counteracted ROS generation and subsequent alteration of monolayer integrity and membrane oxidative damage. The protective action of the extract is likely due to the scavenging ability of its main components, as hydroxytyrosol, oleuropein, and verbascoside among the secoiridoids and derivatives. Since olives phenolic compounds concentrate in the intestinal lumen, they may be a useful tool in the prevention of intestinal disorders related to oxidative damage.
Asunto(s)
Antioxidantes/farmacología , Enterocitos/efectos de los fármacos , Olea/química , Estrés Oxidativo/efectos de los fármacos , Fenoles/farmacología , Extractos Vegetales/farmacología , Células CACO-2 , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Enterocitos/metabolismo , Glutatión/metabolismo , Humanos , Peroxidación de Lípido/efectos de los fármacos , Oxidación-Reducción , Especies Reactivas de OxígenoRESUMEN
The phenolic fraction of extra virgin olive oil (EVOO) concentrates before absorption in the intestinal lumen, where it may contribute to the modulation of enterocytes response to oxidative and inflammatory stimuli. We evaluated the ability of two monovarietal EVOOs phenolic extracts, Bosana and Nera di Gonnos/Tonda di Cagliari, typical and widespread varieties in Sardinia (Italy), to counteract in enterocytes like Caco-2 cells the pro-oxidant action of oxidized lipids, tert-butyl hydroperoxide (TBH) or a mixture of oxysterols of dietary origin. We confirmed that TBH treatment causes a significant increase of ROS production, GSH depletion, increase of MDA, fatty acids hydroperoxides and 7-ketocholesterol, and showed first evidence of oxidative imbalance and cell damage due to oxysterols exposure. Preincubation of cells with the phenolic extracts significantly attenuated oxidative modifications. Bosana extract showed the highest concentration of total phenols, mainly hydroxytyrosol and tyrosol, and was the most active in presence of TBH, where the free radical scavenging activity of these simple phenols seems to be a determining factor. The two extracts were equally effective, in spite of the different composition, in presence of oxysterols, where ROS production probably occurs according to different and more complex mechanisms.
Asunto(s)
Lípidos/efectos adversos , Aceite de Oliva/química , Fenoles/química , Extractos Vegetales/farmacología , Células CACO-2 , Grasas de la Dieta , Humanos , Lípidos/química , Oxidación-Reducción , Extractos Vegetales/químicaRESUMEN
The aim of this study was to investigate the ability of the sulfate metabolites of hydroxytyrosol (HT) and tyrosol (TYR) to act as antioxidants counteracting the pro-oxidant effect of oxidized cholesterol in intestinal cells. For this purpose, we synthesized sulfate metabolites of HT and TYR using a chemical methodology and examined their antioxidant activity in Caco-2 monolayers in comparison with the parent compounds. Exposure to oxidized cholesterol led to ROS production, oxidative damage, as indicated by the MDA increase, a decrease of reduced glutathione concentration and an enhancement of glutathione peroxidase activity. All the tested compounds were able to counteract the oxidizing action of oxidized cholesterol; HT and TYR sulfate metabolites showed an efficiency in protecting intestinal cells comparable to that of the parent compounds, strengthening the assumption that the potential beneficial effect of the parent compounds is retained, although extensive metabolisation occurs, the resulting metabolites being able to exert a biological action themselves.
Asunto(s)
Colesterol/toxicidad , Enterocitos/efectos de los fármacos , Alcohol Feniletílico/análogos & derivados , Células CACO-2 , Colesterol/química , Humanos , Malondialdehído , Estructura Molecular , Oxidantes/toxicidad , Oxidación-Reducción , Alcohol Feniletílico/química , Alcohol Feniletílico/metabolismo , Alcohol Feniletílico/farmacología , Sulfatos/química , Sulfatos/metabolismoRESUMEN
While neurochemical metabolite modifications, determined by different techniques, have been diffusely reported in human and mice brains affected by transmissible spongiform encephalopathies (TSEs), this aspect has been little studied in the natural animal hosts with the same pathological conditions so far. Herein, we investigated, by high resolution (1)H NMR spectroscopy and multivariate statistical data analysis, the brain metabolite profile of sheep exposed to a scrapie agent in a naturally affected flock. On the basis of clinical examinations and western blotting analysis for the pathological prion protein (PrP(Sc)) in brain tissues, sheep were catalogued as not infected (H), infected with clinical signs (S), and infected without clinical signs (A). By discriminant analysis of spectral data, comparing S vs. H, we found a different metabolite distribution, with inosine, cytosine, creatine, and lactate being higher in S than in H brains, while the branched chain amino acids (leucine, isoleucine, and valine), phenylalanine, uracil, tyrosine, gamma-amino butyric acid, total aspartate (aspartate + N-acetyl aspartate) being lower in S. By a soft independent modelling of class analogy approach, 1 out of 3 A samples was assigned to class H. Furthermore, A brains were found to be higher in choline and choline-containing compounds. By means of partial least squares regression, an excellent correlation was found between the PrP(Sc) amount and the (1)H NMR metabolite profile of infected (S and A) sheep, and the metabolite mostly correlated with PrP(Sc) was alanine. The overall results, obtained using different chemometric tools, were able to describe a brain metabolite profile of infected sheep with and without clinical signs, compared to healthy ones, and indicated alanine as a biomarker for PrP(Sc) amounts in scrapie brains.
Asunto(s)
Encéfalo/metabolismo , Metaboloma , Scrapie/metabolismo , Aminoácidos/metabolismo , Animales , Estudios de Casos y Controles , Metabolómica , Proteínas PrPSc/metabolismo , Espectroscopía de Protones por Resonancia Magnética , OvinosRESUMEN
Prion diseases are fatal neurodegenerative disorders affecting many mammals, ovine scrapie being the archetypal prion disease. Several independent studies in murine and cell-based models of scrapie have highlighted the presence of a link between prion generation and lipid alterations; yet, no data on natural disease are available. In this study we investigated levels of total lipids and cholesterol as well as profiles of fatty acids in brain homogenates from symptomatic and asymptomatic scrapie-infected sheep vs. healthy sheep, all belonging to the same flock. Lipid extracts were analyzed by means of gas chromatography and high performance liquid chromatography. Data of fatty acids were submitted to multivariate statistical analysis to give a picture of the brain lipid profiles of sheep. Interestingly, results revealed abnormalities in the brain fatty acid unsaturation of infected/symptomatic animals. Significant reduction of monoene 18:1 n-9 was detected in brain lipids from infected/symptomatic sheep, as compared to healthy and infected/asymptomatic animals, and this alteration occurred in combination with a significant increase in 18:0 level. The unsupervised Principal Component Analysis showed that infected/symptomatic and healthy sheep samples lie in two different regions of the plot, infected/asymptomatic lie mostly next to healthy. The increase of cerebral saturated fatty acids provides a rough indication of presumed alterations in lipid raft domains of nervous cells during scrapie, suggesting that they may exist in a notable viscous liquid-ordered state. Such physicochemical alteration would have a profound impact on the raft thermodynamic properties, its spatial organization, and signal transduction, all potentially relevant for prion generation.
Asunto(s)
Encéfalo/patología , Ácidos Grasos/análisis , Lípidos/análisis , Scrapie/metabolismo , Scrapie/patología , Animales , Encéfalo/metabolismo , Ácidos Grasos/metabolismo , Metabolismo de los Lípidos , OvinosRESUMEN
One of the most important sites of polyphenol action seems to be in the gastrointestinal system before absorption. We investigated the ability of three wine phenolic extracts, obtained from grape varieties grown in Sardinia, Cannonau (red), Vermentino and Malvasia (white), to exert an antioxidant action against tert-butyl hydroperoxide (TBH)-induced oxidative damage to Caco-2 cell monolayers as a model system of the human intestine. TBH treatment caused the disruption of epithelial integrity, measured as transepithelial electrical resistance, and markers of the peroxidation process of membrane lipids, MDA, fatty acid hydroperoxides and 7-ketocholesterol. All wine extracts were able to counteract the oxidising action of TBH and, in spite of the differences in phenolic composition, exerted a comparable activity. Our findings point out a direct antioxidant action of the wine extracts on enterocytes exposed to oxidising species and further support the opinion that total phenolic content is not essential for antioxidant activity.
Asunto(s)
Mucosa Intestinal/metabolismo , Intestinos/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/farmacología , Vitis/química , Vino/análisis , Antioxidantes/farmacología , Células CACO-2 , Humanos , Peroxidación de Lípido/efectos de los fármacos , Polifenoles/farmacologíaRESUMEN
Hydroxytyrosol (2-(3',4'-dihydroxyphenyl)ethanol; HT), the most active ortho-diphenolic compound, present either in free or esterified form in extravirgin olive oil, is extensively metabolized in vivo mainly to O-methylated, O-sulfated and glucuronide metabolites. We investigated the capacity of three glucuronide metabolites of HT, 3'-O-ß-d-glucuronide and 4'-O-ß-d-glucuronide derivatives and 2-(3',4'-dihydroxyphenyl)ethanol-1-O-ß-d-glucuronide, in comparison with the parent compound, to inhibit H(2)O(2) induced oxidative damage and cell death in LLC-PK1 cells, a porcine kidney epithelial cell line. H(2)O(2) treatment exerted a toxic effect inducing cell death, interacting selectively within the pro-death extracellular-signal relate kinase (ERK 1/2) and the pro-survival Akt/PKB signaling pathways. It also produced direct oxidative damage initiating the membrane lipid peroxidation process. None of the tested glucuronides exhibited any protection against the loss in renal cell viability. They also failed to prevent the changes in the phosphorylation states of ERK and Akt, probably reflecting their inability to enter the cells, while HT was highly effective. Notably, pretreatment with glucuronides exerted a protective effect at the highest concentration tested against membrane oxidative damage, comparable to that of HT: the formation of malondialdehyde, fatty acid hydroperoxides and 7-ketocholesterol was significantly inhibited.
Asunto(s)
Antioxidantes/farmacología , Células Epiteliales/efectos de los fármacos , Glucurónidos/farmacología , Peróxido de Hidrógeno/toxicidad , Estrés Oxidativo/efectos de los fármacos , Alcohol Feniletílico/análogos & derivados , Animales , Antioxidantes/síntesis química , Antioxidantes/química , Línea Celular , Células Epiteliales/metabolismo , Glucurónidos/síntesis química , Glucurónidos/química , Cetocolesteroles/metabolismo , Túbulos Renales/citología , Peróxidos Lipídicos/metabolismo , Malondialdehído/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Alcohol Feniletílico/metabolismo , Alcohol Feniletílico/farmacología , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , PorcinosRESUMEN
The importance of n-3 polyunsaturated fatty acid (n-3 PUFA) intake has long been recognized in human nutrition. Although health benefits, n-3 PUFA are subject to rapid and/or extensive oxidation during processing and storage, resulting in potential alteration in nutritional composition and quality of food. Bottarga, a salted and semi-dried mullet ( Mugil cephalus ) ovary product, is proposed as an important source of n-3 PUFA, having high levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). In this work, we investigated the extent of lipid oxidation of grated bottarga samples during 7 months of storage at -20 °C and room temperature under light exposure. Cell viability, lipid composition, and lipid peroxidation were measured in intestinal differentiated Caco-2 cell monolayers after 6-48 h of incubation with lipid and hydrophilic extracts obtained from bottarga samples at different storage conditions. The storage of bottarga did not affect the n-3 PUFA level, but differences were observed in hydroperoxide levels in samples from different storage conditions. All tested bottarga extracts did not show a toxic effect on cell viability of differentiated Caco-2 cells. Epithelial cells incubated with bottarga oil had significant changes in fatty acid composition but not in cholesterol levels with an accumulation of EPA, DHA, and 22:5. Cell hydroperoxides were higher in treated cells, in relation to the oxidative status of bottarga oil. Moreover, the bottarga lipid extract showed an in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells).
Asunto(s)
División Celular/efectos de los fármacos , Productos Pesqueros , Intestinos/química , Intestinos/citología , Lípidos/análisis , Smegmamorpha , Animales , Células CACO-2 , Supervivencia Celular/efectos de los fármacos , Ácidos Grasos , Ácidos Grasos Omega-3/análisis , Femenino , Productos Pesqueros/análisis , Alimentos en Conserva , Humanos , Intestinos/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Lípidos/aislamiento & purificación , Ovario/químicaRESUMEN
This study examines the protective effect of arzanol, a pyrone-phloroglucinol etherodimer from Helichrysum italicum subsp. microphyllum, against the oxidative modification of lipid components induced by Cu(2+) ions in human low density lipoprotein (LDL) and by tert-butyl hydroperoxide (TBH) in cell membranes. LDL pre-treatment with arzanol significantly preserved lipoproteins from oxidative damage at 2h of oxidation, and showed a remarkable protective effect on the reduction of polyunsaturated fatty acids and cholesterol levels, inhibiting the increase of oxidative products (conjugated dienes fatty acids hydroperoxides, 7ß-hydroxycholesterol, and 7-ketocholesterol). Arzanol, at non-cytotoxic concentrations, exerted a noteworthy protection on TBH-induced oxidative damage in a line of fibroblasts derived from monkey kidney (Vero cells) and in human intestinal epithelial cells (Caco-2), decreasing, in both cell lines, the formation of oxidative products (hydroperoxides and 7-ketocholesterol) from the degradation of unsaturated fatty acids and cholesterol. The cellular uptake and transepithelial transport of the compound were also investigated in Caco-2 cell monolayers. Arzanol appeared to accumulate in Caco-2 epithelial cells. This phenol was able to pass through the intestinal Caco-2 monolayers, the apparent permeability coefficients (P(app)) in the apical-to-basolateral and basolateral-to-apical direction at 2h were 1.93±0.36×10(-5) and 2.20±0.004×10(-5)cm/s, respectively, suggesting a passive diffusion pathway. The results of the work qualify arzanol as a potent natural antioxidant with a protective effect against lipid oxidation in biological systems.
Asunto(s)
Antioxidantes/farmacología , Cobre/efectos adversos , Peroxidación de Lípido/efectos de los fármacos , Lipoproteínas LDL/metabolismo , Floroglucinol/análogos & derivados , Pironas/farmacología , terc-Butilhidroperóxido/efectos adversos , Animales , Antioxidantes/farmacocinética , Transporte Biológico , Células CACO-2 , Chlorocebus aethiops , Humanos , Oxidación-Reducción/efectos de los fármacos , Floroglucinol/farmacocinética , Floroglucinol/farmacología , Pironas/farmacocinética , Células VeroRESUMEN
Complex polyphenols present in extravirgin olive oil are not directly absorbed, but undergo gastrointestinal biotransformation, increasing the relative amount of tyrosol (TYR) and hydroxytyrosol (HT) entering the small and large intestine. We investigated the capacity of TYR and HT to inhibit the insult of dietary lipid hydroperoxydes on the intestinal mucosa, using cultures of Caco-2, a cell line with enterocyte-like features, and studying the effect of tert-butyl hydroperoxide (TBH) treatment on specific cell membrane lipid targets. The effect of homovanillic alcohol (HVA), metabolite of HT in humans and detected as metabolite of HT in Caco-2 cells, was also evaluated. Exposure to TBH induced a significant increase of the level of MDA, the formation of fatty acid hydroperoxides and 7-ketocholesterol and the loss of α-tocopherol. Pretreatment with both HT and HVA protected Caco-2 cells from oxidative damage: there was no significant detection of oxidation products and the level of α-tocopherol was preserved. Noteworthy, TYR also exerted a protective action against fatty acids degradation. In vitro trials, where the simple phenols were tested during linoleic acid and cholesterol oxidation, gave evidence of a direct scavenging of peroxyl radicals and suggested a hydrogen atom-donating activity.
Asunto(s)
Antioxidantes/farmacología , Peroxidación de Lípido/efectos de los fármacos , Fenoles/farmacología , Aceites de Plantas/farmacología , Antioxidantes/química , Células CACO-2 , Supervivencia Celular/efectos de los fármacos , Colesterol/química , Ácidos Grasos/química , Depuradores de Radicales Libres/química , Humanos , Cetocolesteroles/química , Ácido Linoleico/química , Malondialdehído/química , Aceite de Oliva , Oxidación-Reducción , Fenoles/química , Alcohol Feniletílico/análogos & derivados , Alcohol Feniletílico/química , Aceites de Plantas/química , alfa-Tocoferol/químicaRESUMEN
The olive oil polyphenol, hydroxytyrosol (HT), is believed to be capable of exerting protection against oxidative kidney injury. In this study we have investigated the ability of HT and its O-methylated metabolite, homovanillic alcohol (HVA) to protect renal cells against oxidative damage induced by hydrogen peroxide. We show that both compounds were capable of inhibiting hydrogen peroxide-induced kidney cell injury via an ability to interact with both MAP kinase and PI3 kinase signalling pathways, albeit at different concentrations. HT strongly inhibited death and prevented peroxide-induced increases in ERK1/2 and JNK1/2/3 phosphorylation at 0.3 microM, whilst HVA was effective at 10 microM. At similar concentrations, both compounds also prevented peroxide-induced reductions in Akt phosphorylation. We suggest that one potential protective effect exerted by olive oil polyphenols against oxidative kidney cell injury may be attributed to the interactions of HT and HVA with these important intracellular signalling pathways.
Asunto(s)
Citoprotección , Quinasas MAP Reguladas por Señal Extracelular/fisiología , Ácido Homovanílico/farmacología , Peróxido de Hidrógeno/toxicidad , Proteínas Quinasas JNK Activadas por Mitógenos/fisiología , Alcohol Feniletílico/análogos & derivados , Proteínas Proto-Oncogénicas c-akt/fisiología , Transducción de Señal/efectos de los fármacos , Animales , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Alcohol Feniletílico/farmacología , PorcinosRESUMEN
Extra virgin olive oil is rich in phenolic compounds which are believed to exert beneficial effects against many pathological processes, including the development of colon cancer. We show that one of the major polyphenolic constituents of extra virgin olive oil, hydroxytyrosol (HT), exerts strong antiproliferative effects against human colon adenocarcinoma cells via its ability to induce a cell cycle block in G2/M. These antiproliferative effects were preceded by a strong inhibition of extracellular signal-regulated kinase (ERK)1/2 phosphorylation and a downstream reduction of cyclin D1 expression, rather than by inhibition of p38 activity and cyclooxygenase-2 (COX-2) expression. These findings are of particular relevance due to the high colonic concentration of HT compared to the other olive oil polyphenols and may help explain the inverse link between colon cancer and olive oil consumption.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Antineoplásicos Fitogénicos/farmacología , Neoplasias del Colon/tratamiento farmacológico , Ciclina D1/antagonistas & inhibidores , Proteína Quinasa 1 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , Alcohol Feniletílico/análogos & derivados , Adenocarcinoma/patología , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Neoplasias del Colon/patología , Humanos , Alcohol Feniletílico/farmacología , Fosforilación , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
We investigated the capacity of hydroxytyrosol (HT), 3,4-dihydroxyphenylethanol, and homovanillic alcohol (HVA), 4-hydroxy-3-methoxy-phenylethanol, to inhibit H(2)O(2) induced oxidative damage in LLC-PK1, a porcine kidney epithelial cell line, studying the effect of H(2)O(2) on specific cell membrane lipid targets, unsaturated fatty acids and cholesterol. Exposure to H(2)O(2) induced a significant increase of the level of MDA together with a disruption of the membrane structure, with the loss of unsaturated fatty acids, cholesterol and alpha-tocopherol, and the formation of fatty acids hydroperoxides and 7-ketocholesterol. Pretreatment with HT protected renal cells from oxidative damage: the level of membrane lipids was preserved and there was no significant detection of oxidation products. HVA exerted a comparable activity, thus both HT and HVA were able to prevent in renal cells the lipid peroxidation process that plays a central role in tubular cell injury.
Asunto(s)
Células Epiteliales/metabolismo , Ácido Homovanílico/farmacología , Peróxido de Hidrógeno/antagonistas & inhibidores , Peróxido de Hidrógeno/toxicidad , Túbulos Renales/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Oxidantes/toxicidad , Alcohol Feniletílico/análogos & derivados , Sustancias Protectoras , Animales , Antioxidantes/farmacología , Colesterol/metabolismo , Células Epiteliales/efectos de los fármacos , Ácidos Grasos no Esterificados/metabolismo , Túbulos Renales/citología , Túbulos Renales/efectos de los fármacos , Células LLC-PK1 , Malondialdehído/metabolismo , Lípidos de la Membrana/metabolismo , Estrés Oxidativo/efectos de los fármacos , Alcohol Feniletílico/farmacología , Serotonina/metabolismo , Porcinos , alfa-Tocoferol/farmacologíaRESUMEN
Myrtle (Myrtus communis L.), a culinary spice and flavouring agent for alcoholic beverages widespread in the Mediterranean area and especially in Sardinia, contains the structurally unique oligomeric non-prenylated acylphloroglucinols, semimyrtucommulone and myrtucommulone A, whose antioxidant activity was investigated during the oxidative modification of lipid molecules implicated in the onset of cardiovascular diseases. Both acylphloroglucinols showed powerful antioxidant properties during the thermal (140 degrees C), solvent-free degradation of cholesterol. Moreover, the pre-treatment with semimyrtucommulone and myrtucommulone A significantly preserved LDL from oxidative damage induced by Cu(2+) ions at 2h of oxidation, and showed remarkable protective effect on the reduction of polyunsaturated fatty acids and cholesterol, inhibiting the increase of their oxidative products (conjugated dienes fatty acids hydroperoxides, 7beta-hydroxycholesterol, and 7-ketocholesterol). Taking into account the widespread culinary use of myrtle leaves, the results of the present work qualify the natural compounds semimyrtucommulone and myrtucommulone A as interesting dietary antioxidants with potential antiatherogenicity.
Asunto(s)
Colesterol/química , Lipoproteínas LDL/química , Myrtus/metabolismo , Oxígeno/química , Floroglucinol/química , Floroglucinol/farmacología , Antioxidantes/química , Química Física/métodos , Cromatografía de Gases , Ácidos Grasos/química , Humanos , Modelos Químicos , Estrés Oxidativo , Extractos Vegetales/farmacología , Solventes/química , TemperaturaRESUMEN
This study investigated the effect of synthetic capsiate, a simplified analogue of capsiate, and vanillyl alcohol on the oxidative stress induced by tert-butyl hydroperoxide (TBH) in a line of fibroblasts derived from monkey kidney (Vero cells). In response to the TBH-mediated oxidative stress, a reduction of the levels of total unsaturated fatty acids and cholesterol was observed, and a rise in the concentrations of conjugated dienes fatty acids hydroperoxides and 7-ketocholesterol. Pretreatment with both synthetic capsiate and vanillyl alcohol preserved Vero cells from oxidative damage and showed a remarkable protective effect on the reduction of the levels of total unsaturated fatty acids and cholesterol, inhibiting the increase of MDA, conjugated dienes fatty acids hydroperoxides, and 7-ketocholesterol. Both compounds were effective against peroxidation of cell membrane lipids induced by TBH, with synthetic capsiate essentially acting as a pro-drug of vanillyl alcohol, its hydrophilic hydrolytic derivative.
Asunto(s)
Alcoholes Bencílicos/farmacología , Capsaicina/análogos & derivados , Estrés Oxidativo/efectos de los fármacos , terc-Butilhidroperóxido/farmacología , Animales , Alcoholes Bencílicos/análisis , Capsaicina/análisis , Capsaicina/farmacología , Muerte Celular/efectos de los fármacos , Chlorocebus aethiops , Colesterol/análisis , Ácidos Grasos Insaturados/análisis , Fibroblastos/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Malondialdehído/análisis , Células VeroRESUMEN
We investigated the anti-proliferative effects of an olive oil polyphenolic extract on human colon adenocarcinoma cells. Analysis indicated that the extract contained hydroxytyrosol, tyrosol and the various secoiridoid derivatives, including oleuropein. This extract exerted a strong inhibitory effect on cancer cell proliferation, which was linked to the induction of a G2/M phase cell cycle block. Following treatment with the extract (50 microg/ml) the number of cells in the G2/M phase increased to 51.82+/-2.69% relative to control cells (15.1+/-2.5%). This G2/M block was mediated by the ability of olive oil polyphenols (50 microg/ml) to exert rapid inhibition of p38 (38.7+/-4.7%) and CREB (28.6+/-5.5%) phosphorylation which led to a downstream reduction in COX-2 expression (56.9+/-9.3%). Our data suggest that olive oil polyphenols may exert chemopreventative effects in the large intestine by interacting with signalling pathways responsible for colorectal cancer development.