ABSTRACT
Recent studies have expanded the appreciation of the roles of oxysterols triggering inflammatory, immune cytotoxic and apoptotic processes, but have not been considered for proteome analysis. A comparative proteomic study in intestinal epithelial cell cultures incubated (60 µM/24 h) with 7keto-cholesterol or 7keto-stigmasterol was performed. The influence of both compounds was studied following the nLC-TripleTOF analysis. Findings were compared to results for control cultures. In the principal component analysis (PCA) of proteome patterns, two components were extracted accounting for 99.8% of the variance in the protein expression. PCA analysis clearly discriminated between the perturbations in the proteome of cell cultures incubated with 7keto-cholesterol and 7keto-stigmasterol. These proteins participate in mitochondrial function, lipid homeostasis, inflammation and immunity and cell proliferation. Remarkable differences between proteome patterns in cell cultures exposed to 7keto-cholesterol and 7keto-stigmasterol affect macrophage migration inhibitory factor, apolipoprotein E, Bcl-2-associated transcription factor and cellular retinoic acid-binding protein. Besides, exposure to 7keto-stigmasterol increased the concentration of ubiquitin-conjugating enzyme E2 and the mitochondrial superoxide dismutase protein. Such findings raise new questions about safety studies and the regulatory potential of oxysterols in the differentiation and function of intestinal and associated immune cells, their response to environmental stimuli and impairment of absorption processes.
Subject(s)
Enterocytes/drug effects , Gene Expression Regulation/drug effects , Ketocholesterols/toxicity , Oxidants/toxicity , Proteome/drug effects , Stigmasterol/analogs & derivatives , Caco-2 Cells , Enterocytes/enzymology , Enterocytes/metabolism , Gene Expression Profiling , Humans , Peptide Mapping , Principal Component Analysis , Proteome/metabolism , RNA, Messenger/metabolism , Stigmasterol/toxicityABSTRACT
INTRODUCTION: Most of the initiation and maintenance pediatric formulas commercialized in Spain contain prebiotics and/or nucleotides aiming at achieving beneficial effects on prevention of different pathologies and immune protection, similar to human breast milk. However, according to the current legislation, its inclusion is optional since sufficient scientific evidences supporting its mandatory inclusion is lacking. OBJECTIVE: To systematically review the scientific evidence from randomized clinical studies with a control group allowing determining the beneficial role for infant health derived from the inclusion of prebiotics and/or nucleotides in pediatric formulas. METHODOLOGY: We looked for and selected both original papers and reviews in Spanish and English language of placebo controlled randomized clinical studies published in the databases MEDLINE/PubMed, Scielo, Science Direct, and Scopus, until October of 2011. RESULTS: We found 43 randomized clinical trials meeting the inclusion criteria. CONCLUSIONS: More long-term randomized studies with higher number of patients, and standardized supplemental amounts and experimental conditions are needed to establish healthy statements with stronger scientific support regarding the addition of a mixture of prebiotics (GOS/FOS) and/or nucleotides in pediatric formulas. The current trend to include them in pediatric formulas may be justified based on the currently available evidence, as well as their safety and their presence in human breast milk.
Subject(s)
Infant Food , Infant Nutritional Physiological Phenomena , Nucleotides , Prebiotics , Female , Humans , Infant , Infant, Newborn , Male , Milk, Human/chemistry , Nucleotides/analysis , Prebiotics/analysis , Randomized Controlled Trials as TopicABSTRACT
Introducción: Una gran mayoría de preparados infantiles de inicio y continuación comercializados en España presentan en su formulación prebióticos y/o nucleótidos, con objeto de lograr efectos beneficiosos en la prevención de diferentes patologías y protección inmunológica, similares a los de la leche materna. No obstante, atendiendo a la reglamentación vigente, su adición es opcional, ya que no existe suficiente evidencia científica que certifique la obligatoriedad de su inclusión. Objetivo: Revisar sistemáticamente la evidencia científica proveniente de estudios clínicos aleatorizados con grupo control que permita determinar el papel beneficioso para la salud de los lactantes derivada de la adición de prebióticos y/o nucleótidos en preparados infantiles. Metodología: Se buscaron y seleccionaron tanto artículos originales como revisiones en inglés y español de estudios clínicos aleatorizados con grupo control, publicados en las bases de datos de MEDLINE/PubMed, Scielo, Science Direct y Scopus, hasta octubre de 2011. Resultados: Se encontraron 43 ensayos clínicos aleatorizados que cumplieron con los criterios de inclusión. Conclusiones: Son necesarios más estudios a largo plazo, con mayor número de individuos y aleatorizados, así como estandarización de las cantidades suplementadas y demás condiciones experimentales, para poder establecer declaraciones saludables con mayor base científica respecto a la adición de la mezcla de prebióticos (GOS/FOS) y/o nucleótidos en preparados infantiles. La tendencia actual a su inclusión en preparados infantiles puede justificarse en base a las evidencias científicas disponibles hasta el momento, así como a su seguridad y a su presencia en la leche materna (AU)
Introduction: Most of the initiation and maintenance pediatric formulas commercialized in Spain contain prebiotics and/or nucleotides aiming at achieving beneficial effects on prevention of different pathologies and immune protection, similar to human breast milk. However, according to the current legislation, its inclusion is optional since sufficient scientific evidences supporting its mandatory inclusion is lacking. Objective: To systematically review the scientific evidence from randomized clinical studies with a control group allowing determining the beneficial role for infant health derived from the inclusion of prebiotics and/or nucleotides in pediatric formulas. Methodology: We looked for and selected both original papers and reviews in Spanish and English language of placebo controlled randomized clinical studies published in the databases MEDLINE/PubMed, Scielo, Science Direct, and Scopus, until October of 2011. Results: We found 43 randomized clinical trials meeting the inclusion criteria. Conclusions: More long-term randomized studies with higher number of patients, and standardized supplemental amounts and experimental conditions are needed to establish healthy statements with stronger scientific support regarding the addition of a mixture of prebiotics (GOS/FOS) and/or nucleotides in pediatric formulas. The current trend to include them in pediatric formulas may be justified based on the currently available evidence, as well as their safety and their presence in human breast milk (AU)
Subject(s)
Humans , Male , Female , Infant, Newborn , Child , Infant Food , Infant Nutritional Physiological Phenomena , Nucleotides/analysis , Prebiotics/analysis , Milk, Human/chemistryABSTRACT
INTRODUCTION: Fruit beverages are commonly supplemented with milk, vitamins and/or minerals in order to improve their healthy effects by providing some bioactive components that can act additively or synergistically against oxidative stress. AIMS: To test whether iron, zinc, and milk added to fruit beverages do not affect the cytoprotective effect against oxidative damage to Caco-2 cells through GSH-related enzymes induction and cell cycle progression preservation, in comparison with non-supplemented fruit beverage. METHODS: Caco-2 cells were incubated 24 h with the bioaccessible fraction (BF) of eight fruit beverages with/without iron and/or zinc, and/or milk, and then challenged with H2O2 (5 mmol L-1 -2 h). Mitochondrial enzyme activities (MTT test), GSH-Rd and GSH-Px enzyme activities, cell cycle progression and caspase-3 activity were measured. RESULTS AND DISCUSSION: Fruit beverages prevented the deleterious effect of H2O2 on cell viability, with almost all samples reaching control basal levels. Only independent iron or zinc supplementation with/without milk exerted positive effects upon GSH-Rd activity. Both minerals with milk, afforded improved preservation of GSH-Px activity. All samples prevented the decrease in the G1 phase of cell cycle induced by H2O2, except iron supplemented samples with/without milk, but none of them avoided the increase in sub-G1 phase. However, this fact was not associated to caspase-3 activity, with a probable positive effect of zinc upon this parameter. CONCLUSION: Mineral and/or milk supplementation of fruit beverages helps in the prevention of oxidative stress in Caco-2 cells based on cell viability maintenance, GSH-related enzymes activation, cell cycle distribution preservation and inhibition of caspase-3 activation.
Subject(s)
Beverages , Fruit/chemistry , Hydrogen Peroxide/antagonists & inhibitors , Hydrogen Peroxide/pharmacology , Milk , Minerals/pharmacology , Oxidative Stress/drug effects , Animals , Caco-2 Cells , Caspase 3/metabolism , Cell Cycle/drug effects , Cell Survival/drug effects , Dietary Supplements , Digestion/physiology , Glutathione/metabolism , Glutathione Peroxidase/metabolism , Glutathione Reductase/metabolism , Humans , Mitochondria/drug effects , Mitochondria/enzymologyABSTRACT
Introduction: Fruit beverages are commonly supplemented with milk, vitamins and/or minerals in order to improve their healthy effects by providing some bioactive components that can act additively or synergistically against oxidative stress. Aims: To test whether iron, zinc, and milk added to fruit beverages do not affect the cytoprotective effect against oxidative damage to Caco-2 cells through GSH-related enzymes induction and cell cycle progression preservation, in comparison with non-supplemented fruit beverage. Methods: Caco-2 cells were incubated 24 h with the bioaccessible fraction (BF) of eight fruit beverages with/without iron and/or zinc, and/or milk, and then challenged with H2O2 (5 mmol L-1 -2 h). Mitochondrial enzyme activities (MTT test), GSH-Rd and GSH-Px enzyme activities, cell cycle progression and caspase-3 activity were measured. Results and discussion: Fruit beverages prevented the deleterious effect of H2O2 on cell viability, with almost all samples reaching control basal levels. Only independent iron or zinc supplementation with/without milk exerted positive effects upon GSH-Rd activity. Both minerals with milk, afforded improved preservation of GSH-Px activity. All samples prevented the decrease in the G1 phase of cell cycle induced by H2O2, except iron supplemented samples with/without milk, but none of them avoided the increase in sub-G1 phase. However, this fact was not associated to caspase-3 activity, with a probable positive effect of zinc upon this parameter. Conclusion: Mineral and/or milk supplementation of fruit beverages helps in the prevention of oxidative stress in Caco-2 cells based on cell viability maintenance, GSH-related enzymes activation, cell cycle distribution preservation and inhibition of caspase-3 activation (AU)
Introducción: En la actualidad las bebidas a base de zumo de frutas llevan adicionadas leche, vitaminas y/o minerales con objeto de mejorar sus efectos beneficiosos para la salud mediante el aporte de numerosos compuestos bioactivos que pueden actuar de forma aditiva o sinérgica frente al estrés oxidativo. Objetivos: Evaluar si la adición de hierro, cinc y leche a las bebidas a base de zumo de frutas no afecta al efecto cito-protector frente al daño oxidativo en células Caco-2 a través de la inducción de enzimas del ciclo del GSH y la preservación de la progresión del ciclo celular, en comparación con la bebida a base de zumo de frutas no suplementada. Métodos: Las células Caco-2 se incubaron 24 h con las fracciones bioaccesibles (FB) de ocho bebidas a base de zumo de frutas con/sin hierro y/o cinc y/o leche, y se sometieron a estrés oxidativo con H2O2 (5 mmol L-1-2 h). Se determinó la actividad enzimática mitocondrial (test MTT), la actividad de las enzimas GSH-Rd y GSH-Px, la progresión del ciclo celular y la actividad de la enzima caspasa-3. Resultados y discusión: Las bebidas a base de zumo de frutas previnieron del efecto perjudicial del H2O2 sobre la viabilidad celular, con casi todas las muestras alcanzando los niveles basales del control. Sólo la adición independiente de hierro o cinc con/sin leche ejerció efectos positivos sobre la actividad de la enzima GSH-Rd. Por otra parte, ambos minerales con leche proporcionaron una mejor preservación en la actividad de la enzima GSH-Px. Todas las muestras previnieron el descenso en la fase G1 del ciclo celular inducido por el H2O2, excepto la muestra adicionada de hierro, pero ninguna de ellas evitó el incremento en la fase subG1 del ciclo celular. Sin embargo, este hecho no estuvo asociado con la actividad de la enzima caspasa-3, con un probable efecto positivo del cinc sobre este parámetro. Conclusión: La adición de minerales y/o leche a las bebidas a base de zumo de frutas ayuda en la prevención de estrés oxidativo en células Caco-2 mediante el mantenimiento de la viabilidad celular, activación de enzimas del ciclo del GSH, preservación de la progresión del ciclo celular e inhibición en la activación de la enzima caspasa-3 (AU)
Subject(s)
Humans , Animals , Beverages , Fruit/chemistry , Hydrogen Peroxide , Hydrogen Peroxide/pharmacology , Milk , Minerals/pharmacology , Oxidative StressABSTRACT
INTRODUCTION: Colorectal cancer risks could be reduced by polyphenol-rich diets that inhibit tumour cell growth. AIMS: To determine the polyphenolic profile of four fruit beverages (FbZn, FbZnFe, FbZnM and FbZnFeM) as affected by the presence of Zn with/without Fe and with/without skimmed milk, and the digestion conditions. To evaluate the antiproliferative activity of bioaccessible fractions against Caco-2 and HT-29 cells. To clarify whether cell cycle arrest and/or apoptosis is involved in their possible antiproliferative activity. METHODS: The polyphenolic profiles were analyzed by RP-HPLC-DAD before and after in vitro gastrointestinal digestion. Cell proliferation and viability were measured using Trypan blue test, mitochondrial enzyme activity by means MTT test, cell cycle distribution using flow cytometry and apoptosis by means Hoechst dye. RESULTS AND DISCUSSION: The presence of zinc, iron and/or milk decreased the soluble extractable phenolic content before digestion probably by chelate formation, FbZn and FbZnFe being the samples with the highest soluble extractable phenolics. After digestion, a decrease in phenolics was observed in all zinc-fortified samples (up to 32% with respect to the original fruit beverages) - the FbZnFeM sample showing the lowest soluble extractable phenolic content, though with the lowest percentage decrease in phenolics (14%). FbZnM digest (approximately 50 microM total soluble extractable phenolics) was the sample that most inhibited Caco-2 and HT-29 cell proliferation after 24 h of incubation, without cytotoxicity. The specific combination of phytochemicals in FbZnM digest proved cytostatic and significantly suppressed proliferation through cell cycle arrest in the S-phase in both cell lines, without apoptosis.
Subject(s)
Beverages/analysis , Flavonoids/analysis , Flavonoids/pharmacology , Food, Fortified/analysis , Fruit , Phenols/analysis , Phenols/pharmacology , Zinc/analysis , Zinc/pharmacology , Caco-2 Cells/drug effects , Cell Proliferation/drug effects , Colonic Neoplasms , HT29 Cells/drug effects , Humans , PolyphenolsABSTRACT
Introduction: Colorectal cancer risks could be reduced by polyphenol-rich diets that inhibit tumour cell growth. Aims: To determine the polyphenolic profile of four fruit beverages (FbZn, FbZnFe, FbZnM and FbZnFeM) as affected by the presence of Zn with/without Fe and with/without skimmed milk, and the digestion conditions. To evaluate the antiproliferative activity of bioaccessible fractions against Caco-2 and HT-29 cells. To clarify whether cell cycle arrest and/or apoptosis is involved in their possible antiproliferative activity. Methods: The polyphenolic profiles were analyzed by RP-HPLC-DAD before and after in vitro gastrointestinal digestion. Cell proliferation and viability were measured using Trypan blue test, mitochondrial enzyme activity by means MTT test, cell cycle distribution using flow cytometry and apoptosis by means Hoechst dye. Results and discussion: The presence of zinc, iron and/or milk decreased the soluble extractable phenolic content before digestion probably by chelate formation, FbZn and FbZnFe being the samples with the highest soluble extractable phenolics. After digestion, a decrease in phenolics was observed in all zinc-fortified samples (up to 32% with respect to the original fruit beverages) - the FbZnFeM sample showing the lowest soluble extractable phenolic content, though with the lowest percentage decrease in phenolics (14%). FbZnM digest (~50 μM total soluble extractable phenolics) was the sample that most inhibited Caco-2 and HT-29 cell proliferation after 24 h of incubation, without cytotoxicity. The specific combination of phytochemicals in FbZnM digest proved cytostatic and significantly suppressed proliferation through cell cycle arrest in the S-phase in both cell lines, without apoptosis (AU)
Introducción: Los riesgos de cáncer colorrectal podrían reducirse mediante dietas ricas en polifenoles, los cuales pueden inhibir el crecimiento de células tumorales. Objetivos: Determinar cómo el perfil polifenólico de cuatro bebidas a base de zumo de frutas (FbZn, FbZnFe, FbZnM and FbZnFeM) puede verse afectado por la presencia de Zn con/sin Fe y con/sin leche desnatada, así como por las condiciones de digestión gastrointestinal. Evaluar la actividad antiproliferativa de las fracciones bioaccesibles en células Caco-2 y HT-29. Averiguar si un arresto en el ciclo celular y/o apoptosis están implicados en su posible actividad antiproliferativa. Métodos: Los perfiles polifenólicos se analizaron mediante RP-HPLC-DAD antes y después de la digestión gastrointestinal in vitro. La proliferación y viabilidad celular se determinaron con el azul tripán, la actividad enzimática mitocondrial por medio del test MTT, la distribución del ciclo celular por citometría de flujo y la apoptosis mediante la sonda fluorescente Hoechst. Resultados y discusión: La presencia de cinc, hierro y/o leche disminuyó el contenido de polifenoles solubles extraíbles antes de la digestión, probablemente por formación de quelatos. Tras la digestión, se observa un descenso en los polifenoles en todas las muestras suplementadas con cinc (hasta un 32% con respecto a las bebidas de frutas originales), siendo la muestra FbZnFeM aquella con menor contenido de polifenoles solubles extraíbles, aunque con el menor porcentaje de descenso de polifenoles (14%). La muestra digerida de FbZnM (~50 μM polifenoles totales solubles extraíbles) fue la que más inhibió la proliferación de las células Caco-2 y HT-29 tras 24 h de incubación, sin citotoxicidad. La combinación específica de compuestos fitoquímicos en la fracción bioaccesible de FbZnM actuó de forma citostática, disminuyendo la proliferación celular de forma significativa mediante arresto del ciclo celular en la fase S en ambas líneas celulares, no acompañado de apoptosis (AU)
