Effects of two highly monounsaturated oils on lipid composition and enzyme activities in rat jejunum

The effects of two monounsaturated fatty acid (MUFA) oils, olive oil (OO)and high-oleic sunflower oil (HOSO), with high content in oleic acid butdiffering in their non-fatty acid fraction, on brush-border membrane(BBM) lipid composition and fluidity and on mucosal enzyme activitiesof rat jejunum were studied. Animals were given semipurified diet withlinoleic acid to prevent essential fatty acid deficiency (control group)or semipurified diet containing 10% of either OO or HOSO for 12weeks. There was a significant decrease in the content of jejunalBBM phospholipids together with an increase in the level of freecholesterol in both oil-fed rats, when compared to controlgroup. Although the increase in the BBM free cholesterol levelwas not statistically significant in HOSO-fed rats, a significantdecrease in the phospholipid/free cholesterol ratio was found inboth OO and HOSO-fed animals compared to control group. Rat jejunalBBM had a high level of free fatty acids which was increased in BBMisolated from OO and HOSO-fed animals. There was no statisticalsignificant difference in the phospholipid distribution between thecontrol and the OO group. However, HOSO-fed animals showed the lowestlevel of phosphatidylethanolamine together with the highestphosphatidylcholine content and the phosphatidylcholine/sphingomyelinratio. The fatty acid pattern of jejunal BBM lipids was modifiedaccording to the major fatty acids in the oils. There was a decreasein both stearic acid (18:0) and linoleic acid (18:2 n-6), togetherwith an increase in oleic acid (18:1 n-9) in jenunal BBM isolatedfrom both oil experimental groups. All these results were accompaniedby a significant increase in the BBM fluidity (as assessed bysteady-state fluorescence polarization of diphenylhexatriene) isolatedfrom oil-fed rat, when compared to control group. OO and HOSO-fedanimals had the lowest activities of sucrase and maltase, whilealkaline phosphatase activity only was decreased in HOSO-fedanimals. The specific activity of maltase was not modified in anyexperimental rats. In summary, both MUFA oils induced similar effectson jejunal BBM lipid composition, fluidity, sucrase, maltase andlactase activities. Furthermore, HOSO intake resulted in a lowestalkaline phosphatase activity which was accompanied by changes inindividual phospholipid composition. All these results suggest thateffects of MUFA oils on jejunal BBM lipid composition and hydrolaseactivities are most likely due to the presence of high content ofoleic acid rather than other components contained in the non-fattyacid of olive oil

Intracellular Chloride Concentration Changes Modulate IL-1β Expression and Secretion in Human Bronchial Epithelial Cultured Cells

Cystic fibrosis (CF) is caused by mutations in the CFTR gene, which encodes a cAMP‐regulated chloride channel. Several cellular functions are altered in CF cells. However, it is not clear how the CFTR failure induces those alterations. We have found previously several genes differentially expressed in CF cells, including c‐Src, MUC1, MTND4, and CISD1 (CFTR‐dependent genes). Recently, we also reported the existence of several chloride‐dependent genes, among them GLRX5 and RPS27. Here, varying the intracellular chloride concentration [Cl−]i of IB3‐1 CF bronchial epithelial cells, we show that IL‐1β mRNA expression and secretion are also under Cl− modulation. The response to Cl− is biphasic, with maximal effects at 75 mM Cl−. The regulation of the IL‐1β mRNA expression involves an IL‐1β autocrine effect, since in the presence of the IL‐1β receptor antagonist IL1RN or anti‐IL‐1β blocking antibody, the mRNA response to Cl− disappeared. Similar effects were obtained with the JNK inhibitor SP600125, the c‐Src inhibitor PP2 and the IKK inhibitor III (BMS‐345541). On the other hand, the IL‐1β secretion is still modulated by Cl− in the presence of IL‐1RN, IL‐1β blocking antibody, or cycloheximide, suggesting that Cl− is affecting the IL‐1β maturation/secretion, which in turn starts an autocrine positive feedback loop. In conclusion, the Cl− anion acts as a second messenger for CFTR, modulating the IL‐1β maturation/secretion. The results also imply that, depending on its intracellular concentration, Cl− could be a pro‐inflammatory mediator.Fil: Clauzure, Mariangeles. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Valdivieso, Ángel Gabriel. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Massip Copiz, María Macarena. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Mori, Consuelo. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; ArgentinaFil: Dugour, Andrea Vanesa. Fundación Pablo Cassara; ArgentinaFil: Figueroa, Juan Manuel. Fundación Pablo Cassara; ArgentinaFil: Santa Coloma, Tomás Antonio. Pontificia Universidad Católica Argentina "Santa María de los Buenos Aires". Instituto de Investigaciones Biomédicas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas; Argentin

Obtaining from grape pomace an enzymatic extract with anti-inflammatory properties

Grape pomace, a winemaking industry by-product, is a rich source of bioactive dietary compounds. Using proteases we have developed an enzymatic process for obtaining a water-soluble extract (GP-EE) that contains biomolecules such as peptides, carbohydrates, lipids and polyphenols in soluble form. Of especial interest is its high polyphenol content (12 %), of which 77 % are flavonoids and 33 % are phenolic acids. The present study evaluates in vitro the potential anti-inflammatory effect of GP-EE by monitoring the expression of inflammatory molecules on N13 microglia cells stimulated with lipopolysaccharide (LPS). GP-EE decreases the mRNA levels of the inflammatory molecules studied. The molecules under study were as follows: inducible nitric oxide synthase (iNOS), tumor necrosis factor- α (TNF-α), interleukin-1β (IL-1β), the ionized calcium binding adaptor molecule-1(Iba-1) and the Toll like receptor-4 (TLR-4), as well as the iNOS protein level in LPS-stimulated microglia. Our findings suggest that, as a result of its ability to regulate excessive microglial activation, GP-EE possesses antiinflammatory properties. Therefore, acting as a chemopreventive agent, it may be of therapeutic interest in neurodegenerative diseases involving neuroinflammation. We can, therefore, propose GP-EE as a useful natural extract and one that would be beneficial to apply in the field of functional food

NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) modulation by intracellular Cl– concentration

The impairment of the cystic fibrosis transmembrane conductance regulator (CFTR) activity induces intracellular chloride (Cl–) accumulation. The anion Cl–, acting as a second messenger, stimulates the secretion of interleukin-1β (IL-1β), which starts an autocrine positive feedback loop. Here, we show that NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) are indirectly modulated by the intracellular Cl– concentration, showing maximal expression and activity at 75 mM Cl–, in the presence of the ionophores nigericin and tributyltin. The expression of PYD and CARD domain containing (PYCARD/ASC) remained constant from 0 to 125 mM Cl–. The CASP1 inhibitor VX-765 and the NLRP3 inflammasome inhibitor MCC950 completely blocked the Cl–-stimulated IL-1β mRNA expression and partially the IL-1β secretion. DCF fluorescence (cellular reactive oxygen species, cROS) and MitoSOX fluorescence (mitochondrial ROS, mtROS) also showed maximal ROS levels at 75 mM Cl–, a response strongly inhibited by the ROS scavenger N-acetyl-L-cysteine (NAC) or the NADPH oxidase (NOX) inhibitor GKT137831. These inhibitors also affected CASP1 and NLRP3 mRNA and protein expression. More importantly, the serum/glucocorticoid regulated kinase 1 (SGK1) inhibitor GSK650394, or its shRNAs, completely abrogated the IL-1β mRNA response to Cl– and the IL-1β secretion, interrupting the autocrine IL-1β loop. The results suggest that Cl– effects are mediated by SGK1, in which under Cl– modulation stimulates the secretion of mature IL-1β, in turn, responsible for the upregulation of ROS, CASP1, NLRP3 and IL-1β itself, through autocrine signalling.Fil: Clauzure, Mariangeles. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentina. Universidad Nacional de La Pampa; ArgentinaFil: Valdivieso, Ángel Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Dugour, Andrea Vanesa. Fundación Pablo Cassará; ArgentinaFil: Mori, Consuelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Massip Copiz, María Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Aguilar, María Á.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Sotomayor, Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Asensio, Cristian Jorge Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; ArgentinaFil: Figueroa, Juan M.. Fundación Pablo Cassará; ArgentinaFil: Santa Coloma, Tomás Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Ciencias Biológicas y Biomédicas del Sur. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Instituto de Ciencias Biológicas y Biomédicas del Sur; Argentin

Alteraciones morfofuncionales provocadas en el riñón e hígado de rata tras la administración del pesticida atrazina

Los Pesticidas o Plaguicidas han supuesto un gran adelanto en la Agricultura, sin embargo, y ya que su utilización es cada día más frecuente, se hace obligatorio un conocimiento más profundo de los mismos encaminado sobre todo a determinar la posible toxicidad que de una forma directa o indirecta puede tener sobre la humanidad. Aunque se han descrito numerosos casos de intoxicaciones agudas por estos productos son más frecuentes las intoxicaciones crónicas, debido a su estabilidad en el medio ambiente y alimentos y a la facilidad con que se acumulan en los tejidos humanos. Existe no obstante, hoy en día poca información sobre los efectos tóxicos que producen la mayoría de los pesticidas y la repercusión de su acumulación en el organismo sobre estados fisiológicos y patológicos. Por todo ello, y dado que Sevilla es la segunda provincia de España en cuanto a la utilización de Plaguicidas (252), se está llevando a cabo en el Instituto de Investigaciones Científicas “Rector González García” trabajos sobre el efecto de pesticidas a distintos niveles, tanto sobre vegetales, estudiando las modificaciones del índice mitótico en las células meristemáticas, como en animales, dilucidando la acción de diversos pesticidas sobre los distintos órganos. El presente trabajo estudia las alteraciones en riñón e hígado producidas por la atrazina, herbicida que en la actualidad está teniendo un gran auge en Agricultura. Actúa sobre los vegetales inhibiendo la fotosíntesis, si bien, sus efectos tóxicos sobre mamíferos son aún poco conocidos. Se sabe que tiene tendencia a acumularse principalmente en el tejido renal y hepático. En este sentido y con objeto de aportar nuevos datos sobre su toxicidad hemos evaluado las alteraciones morfofuncionales de este pesticida a diversas dosis subletales sobre los órganos dosis subletales sobre los órganos de excreción y metabolización por excelencia

Effects of dietary fats (fish, olive and high-oleic-acid sunflower oils) on lipid composition and antioxidant enzymes in rat liver

The effects of two oleic-acid-rich diets (containing olive oil, OO, and high-oleic-acid sunflower oil, HOSO) on plasma and liver lipid composition detoxification enzyme activities, were compared with those of a fish-oil (FO) diet and a control diet. Compared with the control diet, plasma and hepatic total triacylglycerol concentrations were increased in the animals fed on the HOSO and OO diets and decreased in those fed on the FO diet. The animals fed on FO showed the highest level of cholesterol in the liver and had lower plasma cholesterol concentrations when compared with those fed on the two oleic-acid-rich diets. In comparison with the animals fed on the diets enriched in oleic acid, the FO group showed higher hepatic levels of polyunsaturated fatty acids of the n - 3 series and lower levels of fatty acids of the n - 6 series. Livers of FO-fed rats, compared with those of OO- and HOSO-fed rats showed: (1) significantly higher activities of catalase (EC 1.11.1.6) glutathione peroxidase (EC 1.11.1.9) and Cu/Zn superoxide dismutase (EC 1.15.1.1); (2) no differences in the NADPH-cytochrome c reductase (EC 1.6.99.3) activity. The HOSO diet had a similar effect on liver antioxidant enzyme activities as the OO diet. In conclusion, it appears that changes in the liver fatty acid composition due mainly to n - 3 lipids may enhance the efficiency of the antioxidant defence system. The two monounsaturated fatty acids oils studied (OO and HOSO), with the same high content of oleic acid but different contents of natural antioxidants, had similar effects on the antioxidant enzyme activities measured.This study was supported by Grant ALI96-0456 from the Comision Interministerial de Ciencia y Tecnología, Spain.Peer Reviewe

Liver lipid composition and antioxidant enzyme activities of spontaneously hypertensive rats after ingestion of dietary fats (fish, olive and high-oleic sunflower oils)

Hypertension is associated with greater than normal lipoperoxidation and an imbalance in antioxidant status, suggesting that oxidative stress is important in the pathogenesis of this disease. Although many studies have examined the effect of antioxidants in the diet on hypertensión and other disorders, less attention has been given to the evaluation of the role of specific dietary lipids in modulating endogenous antioxidant enzyme status. Previously, we have described that liver antioxidant enzyme activities may be modulated by consumption of different oils in normotensive rats. The purpose of the present study was to examine the effects of feeding different lipidic diets (olive oil, OO, high-oleic-acid sunflower oil, HOSO, and fish oil, FO) on liver antioxidant enzyme activities of spontaneously hypertensive rats (SHR). Plasma and liver lipid composition was also studied. Total triacylglycerol concentration increases in plasma and liver of animals fed on the HOSO and OO diets and decreases in those fed on the FO diet, relative to rats fed the control diet. The animals fed on the oil-enriched diet show similar hepatic cholesterol and phospholipid contents, which are higher than the control group. Consumption of the FO diet results in a decrease in the total cholesterol and phospholipid concentration in plasma, compared with the high-oleic-acid diets. In liver, the FO group show higher levels of polyunsaturated fatty acids (PUFA) of the (n - 3) series, in relation to the animals fed on the diets enriched in oleic acid. Livers of FO-fed rats, compared with those of OO- and HOSO-fed rats showed: (i) significantly higher activities of catalase, glutathione peroxidase and Cu/Zn superoxide dismutase; (ii) no differences in the NADPH-cytochrome c reductase activity. The HOSO diet had a similar effect on liver antioxidant enzyme activities as the OO diet. In conclusion, it appears that changes in the liver fatty acid composition due mainly to n - 3 lipids may enhance the efficiency of the antioxidant defence system and may yield a benefit in the hypertension status. The two monounsaturated fatty acids oils studied (OO and HOSO), with the same high content of oleic acid, but different content of natural antioxidants, had similar effects on the antioxidant enzyme activities studied.The present study was supported by a grant (AL199-0863) from the Comisión Interministerial de Ciencia y Tecnología, Spain.Peer reviewe