Biomedical subjects

L Arola

Publications and source records attributed to L Arola.

At least 19 recordsLinked to original sources

PubMed2005

Metabolic fate of glucose on 3T3-L1 adipocytes treated with grape seed-derived procyanidin extract (GSPE). Comparison with the effects of insulin.

In this paper we investigate the effects of a grape seed procyanidin extract (GSPE) on the metabolic fate of glucose in adipocytes. Differentiated 3T3-L1 cells were treated with 140 mg/L GSPE or 100 nM insulin for a short period (1 h, acute treatment) or for a long period (15 h, chronic treatment). 2-Deoxy-[1-(3)H]glucose uptake and [1-(14)C]glucose incorporation into cells, glycogen, and lipid were measured. We found that GSPE mimicked the anabolic effects of insulin but there were several important differences. GSPE stimulated glycogen synthesis less than insulin. After chronic exposure, GSPE induced a higher incorporation of glucose into lipid, mainly due to the increase in glucose directed to glycerol synthesis. Our main conclusions, therefore, are that GSPE has insulinomimetic properties and activates glycogen and lipid synthesis. However, the differences between the effects of GSPE and the effects of insulin indicate that GSPE uses mechanisms complementary to those of insulin signaling pathways to bring about these effects.

3T3-L1 Cells↗

PubMed2005

Grape-seed derived procyanidins interfere with adipogenesis of 3T3-L1 cells at the onset of differentiation.

OBJECTIVE: Our group's previous results on the effects of a grape seed procyanidin extract (GSPE) on adipose metabolism showed that peroxisome proliferator-activated receptor-gamma (PPARgamma) plays a central role in the lipolytic effects of GSPE on adipocytes. Since PPARgamma2 is a main regulator of the differentiation process of adipocytes, we investigated whether GSPE affects the adipogenesis of 3T3-L1 cells. DESIGN: We performed a time point screening by treating 3T3-L1 cells with GSPE during the differentiation process for 24 h. MEASUREMENTS: Differentiation markers and differential gene expression due to GSPE treatment (using the microarray technique). RESULTS: Twenty four hour-GSPE treatment at the onset of differentiation reduces adipose-specific markers and maintains the expression of preadipocyte marker preadipocyte factor-1 (Pref-1) significantly elevated. These effects were not found in other time points. Microarray analysis of gene expression after GSPE treatment at the early stage of differentiation showed a modified gene expression profile in which cell cycle and growth-related genes were downregulated by GSPE. CONCLUSION: These results suggest that GSPE affects adipogenesis, mainly at the induction of differentiation, and that procyanidins may have a new role in which they impede the formation of adipose cells.

3T3-L1 Cells↗

PubMed2004

Grape seed-derived procyanidins have an antihyperglycemic effect in streptozotocin-induced diabetic rats and insulinomimetic activity in insulin-sensitive cell lines.

Flavonoids are functional constituents of many fruits and vegetables. Some flavonoids have antidiabetic properties because they improve altered glucose and oxidative metabolisms of diabetic states. Procyanidins are flavonoids with an oligomeric structure, and it has been shown that they can improve the pathological oxidative state of a diabetic situation. To evaluate their effects on glucose metabolism, we administered an extract of grape seed procyanidins (PE) orally to streptozotocin-induced diabetic rats. This had an antihyperglycemic effect, which was significantly increased if PE administration was accompanied by a low insulin dose. The antihyperglycemic effect of PE may be partially due to the insulinomimetic activity of procyanidins on insulin-sensitive cell lines. PE stimulated glucose uptake in L6E9 myotubes and 3T3-L1 adipocytes in a dose-dependent manner. Like insulin action, the effect of PE on glucose uptake was sensitive to wortmannin, an inhibitor of phosphoinositol 3-kinase and to SB203580, an inhibitor of p38 MAPK. PE action also stimulated glucose transporter-4 translocation to the plasma membrane. In summary, procyanidins have insulin-like effects in insulin-sensitive cells that could help to explain their antihyperglycemic effect in vivo. These effects must be added to their antioxidant activity to explain why they can improve diabetic situations.

3T3-L1 Cells↗

PubMed2003

Effect of phenolic compounds on the co-metabolism of citric acid and sugars by Oenococcus oeni from wine.

AIMS: The goal of this study was to examine the growth of Oenococcus oeni in the presence of phenolic compounds under wine conditions and to see how these compounds affect bacterial metabolism. METHODS AND RESULTS: Phenolic compounds have been added to a basal medium that simulates the composition of wine. Fifty milligrams per litre or more of phenolic compounds stimulated bacterial growth. Oenococcus oeni seemed to use citric acid and trehalose, if they were present, before glucose and fructose. Citrate was completely exhausted in three days and the yield of acetate was higher when phenolic compounds were present. CONCLUSIONS: Phenolic compounds reduced the rate of sugar consumption and enhanced citric acid consumption, increasing the yield of acetic acid. SIGNIFICANCE AND IMPACT OF THE STUDY: This study allows a better knowledge of co-metabolism of citric acid and sugars by O. oeni in the presence of phenolic compounds of wine.

Carbohydrate Metabolism↗

PubMed2001

Nonalcoholic components in wine reduce low density lipoprotein cholesterol in normocholesterolemic rats.

Using an experimental model that enables the effects of alcohol to be distinguished from the effects of the nonalcoholic components present in wine, we determined whether wine has effects other than those of alcohol on the metabolism of cholesterol. Male rats were fed a standard diet and had free access to water and either wine or an equivalent alcohol solution for 45 d or 6 mon. Alcohol intake was similar in the two groups of animals. Consumption of the alcohol solution or wine did not influence plasma cholesterol or high density lipoprotein-cholesterol. At 45 d, the consumption both of wine and of alcohol solution reduced low density lipoprotein (LDL)-cholesterol and very low density lipoprotein cholesterol. At 6 mon, only the rats that consumed wine had reduced LDL-cholesterol. After 45 d of consuming alcohol solution, total cholesterol in the aorta was significantly increased mainly as a result of the rise in free cholesterol. In the aorta, the effect of wine consumption was similar to the effect of alcohol solution consumption, although it was less intense. The only clear effect that could be ascribed to the nonalcoholic components in wine was that the LDL-cholesterol was reduced in the long term, although aortic cholesterol was not.

Animals↗

PubMed2000

Effects of copper exposure upon nitrogen metabolism in tissue cultured Vitis vinifera.

The present study analyses the effects of copper treatment on nitrogen metabolism in a closed system. Sauvignon grapevines were cultured in agar and exposed to copper levels ranging from 0.07 (control) to 5 µg Cu g(-1) medium. Ammonium, nitrate, nitrite, individual and total amino acids and protein contents were determined in root, and leaves. Enzyme activities of nitrate and nitrite reductases, glutamine synthetase, glutamate synthase and glutamate dehydrogenase were also determined. Copper exposure produces a dramatic change in nitrogen metabolism, with a reduction of total nitrogen, which reflects the reduction on nitrate and free amino acid contents in both root and leaves. The assimilation of nitrate the main nitrogen source in the medium, requires nitrate reductase, which is reduced to negligible activity as response to copper exposure. Primary nitrogen metabolism is also reduced in leaves, although to a lesser extent than in roots, which may explain the differences between the two organs in response to copper exposure. An alternative system for assimilation of nitrogen through glutamate dehydrogenase in roots is proposed, while higher levels of ammonium and glutamine may fullfil the needs of organic nitrogen in the leaves.

Journal Article↗

PubMed2000

Changes in lipolysis and hormone-sensitive lipase expression caused by procyanidins in 3T3-L1 adipocytes.

OBJECTIVE: To find out whether lipid stores are influenced by phenolic compounds in wine. DESIGN: Differentiated 3T3-L1 cells were treated with catechin, epicatechin or procyanidin extracts with different degrees of polymerization at 150 microM for different periods of time (0.5-24 h). SUBJECTS: Cell line 3T3-L1. MEASUREMENTS: Cellular viability, glycerol-3-phosphate dehydrogenase activity, glycerol release in the medium, HSL mRNA levels, triacylglycerols and protein. RESULTS: Catechin, epicatechin and procyanidin extracts were not toxic for the 3T3-L1 cells in the conditions assayed. Glycerol-3-phosphate dehydrogenase activity was markedly decreased by 150 microM procyanidin extracts. The release of glycerol into the medium was increased in 150 microM procyanidin extract-treated cells and reached a plateau after 15 h exposure. Procyanidins caused a time-dependent reduction in the HSL mRNA levels. CONCLUSIONS: These results suggest that procyanidins from grape and wine affect lipid metabolism whilst their monomers (catechin and epicatechin) do not. This effect is more pronounced when the degree of polymerization is higher. Procyanidin extracts cause a time-dependent reduction in the HSL mRNA levels, inhibit triacylglycerol synthesis and also favour triacylglycerol hydrolysis until the HSL mRNA had reached very low levels.

3T3 Cells↗

PubMed2000

Influence of phenolic compounds on the physiology of Oenococcus oeni from wine.

This study shows that the growth of Oenococcus oeni CECT 4100 in a synthetic medium is affected by phenolic compounds in different ways, depending on their type and concentration. Generally they have no effects at low concentrations, but hydroxycinnamic acids are inhibitory at high concentrations. Malolactic fermentation was stimulated in the presence of catechin and quercetin, but increasingly delayed with increasing amounts of p-coumaric acid. Gallic acid appeared to delay or inhibit the formation of acetic acid from citric acid. This could lead to a better control of malolactic fermentation and suppress the increase in volatile acidity, which is undesirable in the wine-making process.

Acetic Acid↗

PubMed1999

Moderate red wine consumption protects the rat against oxidation in vivo.

The effect of the moderate consumption of red wine on the antioxidant system in rat liver, kidney and plasma has been evaluated. Wistar rats were treated in separate groups as follows: control; red wine for 45 days or 6 months; and 13.5% ethanol for 45 days or 6 months. The consumption of alcoholic beverages was free because the rat could always choose between the alcoholic beverage and the water. In liver, red wine ingestion resulted in higher hepatic superoxide dismutase and glutathione peroxidase activities after 45 days of treatment. The data indicate that wine and ethanol ingestion resulted in lower hepatic malondialdehyde and enhanced hepatic catalase activity in both of the periods studied. In kidney, the reduced glutathione/oxidized glutathione ratio was higher after 45 days of wine consumption, and the malondialdehyde was lower after 6 months of wine consumption. In plasma, malondialdehyde was lower after 6 months of both treatments, but plasmatic vitamin E was higher after red wine consumption while it was lower after ethanol consumption for this period of time. The present study shows that the moderate and prolonged consumption of red wine is consistent with higher protection against oxidation in vivo.

Animals↗

PubMed1999

Ammonium uptake and urea production in hepatocytes from lean and obese Zucker rats.

The metabolic differences in vitro between genetic and dietary obese rats in the uptake of ammonium and amino acids by the liver and their use for ureogenesis have been assayed using hepatocytes isolated from Lean, Obese Zucker (Genetic obese) rats and Dietary obese rats. The hepatocytes of genetic obese animals took up more ammonium and produced higher amounts of urea from ammonium and alanine than those of lean and dietary obese groups (2 and 5 times more respectively). In the lean and dietary obese groups urea synthesis accounted for almost all the nitrogen taken up as ammonium. Thus, dietary and genetic obesity show a widely different handling of nitrogen, and the genetic obese rats need to break down protein to maintain their hepatocyte function.

Alanine↗

PubMed1997

Model for voluntary wine and alcohol consumption in rats.

It has been suggested that moderate consumption of ethanol and wine has a protective effect on human health. Animal models used to date for alcohol consumption can not mimic real situations in humans because the consumption is forced and/or excessive. The present study proposes to determine the effects of a voluntary and ad lib consumption model more similar to that of human behavior. Male Wistar rats had free access to either standard diet and water or the same diet plus red wine, sweet wine, or a solution equivalent to red wine (13.5% ethanol) or to sweet wine (20% ethanol + 130 g/L sucrose) for 30 days or 6 months. Daily wine consumption was 15.8 +/- 0.9 and 2.0 +/- 0.2 ml/day for sweet and red wines, respectively. The consumption of each of the alcoholic solutions was similar to that of the wine they were simulating. Drinking wine or ethanol did not affect food and water intakes or growth rate. Plasma metabolites were not substantially affected by consumption of wine or ethanol. Although moderate and high wine consumption did not change the activity of plasma marker enzymes of tissue damage, the consumption of the 2 alcoholic solutions caused a long-term increase in the activity of aspartate aminotransferase. It seems that wine consumption protects the organism from hepatic lesions induced by ethanol alone.

Alcohol Drinking↗

PubMed1997

Regulation of ammonia-metabolizing enzymes expression in the liver of obese rats: differences between genetic and nutritional obesities.

OBJECTIVE: To determine the expression of carbamoylphosphate synthetase (CPS) and glutamine synthetase (GS) in two different models of obese rats: genetically obese rats and diet obese rats. SUBJECTS: Lean and genetically obese (fa/fa) Zucker rats were used. DESIGN: Lean animals (30-60 d old) were fed for 30 d with standard chow pellets or with a hypercaloric cafeteria diet. Genetically obese rats were fed with standard chow pellets. MEASUREMENTS: Enzyme activity, protein (Western blot) and mRNA (Northern blot) contents of CPS and GS were measured in liver homogenates. RESULTS: In genetically obese animals CPS mRNA content was higher, and GS mRNA content was lower than in control animals; CPS protein content did not change and CPS activity was lower than in control rats. Diet-obese rats had higher levels of CPS and GS mRNAs than control animals; GS protein content and activity was higher than in the control group and at the same time, CPS activity was very low. CONCLUSIONS: In the genetically obese animals the expression of CPS and GS is mainly regulated at the pre-translational level, whereas in the diet obese rats there is a noticeable post-translational component. A reciprocal regulation between CPS and GS can be established at pre-translational levels, whereas at post-transcriptional levels it cannot. It can be concluded that in diet-obese animals the mechanisms involved in retaining nitrogen (low CPS activity) are modulated at the post-translational level.

Ammonia↗

PubMed1997

Muscle amino acid pattern in obese rats.

OBJECTIVE: To determine how the ability of skeletal muscle to manage amino acids is conditioned by obesity. The test was performed in two different models of obese rats: diet-obese rats and genetically obese rats. SUBJECTS: Lean and genetically obese (fa/fa) male Zucker rats were used. DESIGN: For up to 60 d of life lean animals were fed with standard chow pellet or with a hypercaloric cafeteria diet. Genetically obese rats were fed with standard chow pellet during the same period. MEASUREMENTS: Amino acid concentration in the femoral artery and vein and leg blood flow were measured. Free amino acid concentration, protein and nitrogen content and enzyme activities were determined in whole leg muscle. RESULTS: Amino acid availability was increased in diet-obese animals and remained unchanged in the genetically obese group. The genetically obese groups had a lower protein content and unchanged concentration of amino acids in leg muscle. Furthermore, total nitrogen remained unchanged in these animals and they showed an increased activity in alanine aminotransferase and glutamine synthetase. The leg muscle of the diet-obese rats took up several amino acids (Ala, Arg and Val) or released others (mainly Gln and Gly). Conversely, genetically obese rats took up many amino acids and did not release any. CONCLUSIONS: We conclude that in nutritionally obese rats there is an increased availability of amino acids in skeletal muscle. This augmented availability propitiates the increase in the uptake of many amino acids. In genetically obese rats, the lack of variation in amino acid availability points to a possible hereditary alteration that increases the capacity of different amino acid transport systems. Furthermore, the diminished protein content of fa/fa muscle is not due to a lower availability and intracellular pool of amino acids.

Amino Acids↗

PubMed1997

Amino acid metabolism in the kidneys of genetic and nutritionally obese rats.

The ability of the kidney to take up and/or release amino acids has been determined in two models of obesity in Zucker rats, one genetic and the other nutritional (diet-obese). There was a noticeable increase in gluconeogenic amino acids in the arterial blood of diet-obese animals whereas the genetically obese rats showed small variations in the levels of these amino acids. There were significant decreases in renal Gly and Ser, only in the genetically obese rats. Genetically obese animals showed an increase in Glutamine synthetase activity. The uptake and/or release of amino acids showed important variations between the groups. The diet-obese group exhibited greater variation, since this group took up Glu, Ala, Gy, Phe and Citrulline and released Gln, Ser, Arg and Tyr. Genetically obese rats took up Gln, His and Taurine and released Ser. These different patterns may be related to variations in the whole body metabolic rate, since the diet-obese group was more active than the genetically obese group.

Alanine Transaminase↗

PubMed1995

beta-Glucosidase families revealed by computer analysis of protein sequences.

This computational study is a summary of how cloned beta-glucosidase subfamilies are organized. Computations were carried out using General Computer Group, Inc. (GCG) package programs. Twenty-two beta-glucosidases belonging to either cellulolytic or non-cellulolytic organisms were identified. The multialignment of a whole beta-glucosidase family is shown. Two sub-families, A and B, were clearly seen to exist. Sub-family A is further subdivided into sub-families A1 and A2. A1 includes vegetal beta-glucosidases and A2 includes prokaryotic enzymes. Sub-family B has three new sub-families, B1, B2, and B3. The enzymes in B2 are of yeast and/or fungi. Aspartic (D), glutamic (E) and histidine (H) residues, which are thought to be a part of the mechanism of the enzymatic hydrolysis are conserved. The well conserved amino acid sequences of the sub-family A are ITENGA; QUIEGA; HVD; and NEP. The well conserved amino acid sequences of the sub-family B are: SDW; and YN(R,K)(V,L)N.

Amino Acid Sequence↗

PubMed1994

Splanchnic amino acid pattern in genetic and dietary obesity in the rat.

The study of intestinal and hepatic uptake of amino acids by obese rats has been the main objective of this work. The obese animals used were either from genetic or from nutritional basis. In fed state, the intestinal release of amino acids was higher in obese animals than in lean ones (around the double values), but nutritionally and genetically obese rat showed a related pattern, specially for the case of alanine (increased release in relation to controls by a factor of 10). The higher alanine release by intestine is not reversed by 12-h food deprivation. The hepatic availability was also higher in obesity models than in lean animals (increases over 30%). However, the hepatic uptake was increased in genetically obese animals (more than 35%) and decreased in nutritionally obese animals (more than 40%), especially due to alanine uptake (2419, 1100 and 3794 nmols/min/g protein in lean, Diet-ob and fa/fa animals respectively). In obese animals the food deprivation tended to normalize the hepatic uptake of alanine. The differences in alanine uptake between both types of obesity may reflect the differences of urea synthesis.

Amino Acids↗

PubMed1994

Respiratory toxicity of copper.

Respiratory toxicity of copper was tested in Wistar rats by spraying copper sulfate (330 g/l spray) for daily periods of 1 hr in a self-contained chamber for up to 10 days. The respiratory toxicity was compared with that from intraperitoneal administration of 1 mg Cu/mg body weight and with adequate control rats. Analysis of tissue Cu and Zn was done in lung, liver, kidney, and plasma by using atomic absorption spectrophotometry. Similar organ and subcellular distribution of both elements were found between the two treated groups, and only statistically significant higher levels of Cu were found in plasma and liver. After exposure, Cu and Zn were basically associated with a low-molecular-weight component, which eluted as metallothionein in the postmicrosomal fractions.

Administration, Inhalation↗

PubMed1994

Plasma amino acids in hyperphagic pups subjected to a glucose gavage.

An oral gavage of either glucose or saline was given to pups fed either standard diet or cafeteria diet. The plasma amino acid concentrations were measured by a radiochemical method. In the standard diet group, plasma Asn+Asp, Thr, Pro, Cit, Trp and Phe levels were higher in rats receiving a glucose solution than in those given saline solution; taurine (on day 20) and Ser (on day 30) showed also higher plasma values. Plasma Arg and taurine levels in rats receiving glucose were lower than those in rats receiving saline when these pups were fed the cafeteria diet. Tyr (on day 20) and Gly and Pro (on day 30), showed decreased plasma values. The diet consumed during the days preceding a glucose gavage may have pronounced effects on several metabolites, particularly on nitrogen metabolism. The homeostasis of plasma amino acids was held highly constant in spite of the variety of diets supplied, indicating a remarkable homeostatic capacity on amino acidemia against dietetic manipulation.

Administration, Oral↗