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The broiler industry supplies high-quality animal protein to the world. The ban of antibiotics as growth promoters has opened the way for plenty of phytochemicals and antioxidants to be explored. This study summarizes the use of natural antioxidants in a broiler diet as a way through which to deal with stressors, as well as their effects on the expression of various genes. The transcriptional factors and genes involved in the regulation of redox homeostasis are described and emphasis is placed on nuclear factor erythroid 2-related factor 2 and nuclear factor kappa B. Sources such as fruits, vegetables, spices, mushrooms, and algae contain numerous natural antioxidant compounds. The antioxidant activity of these compounds has also been confirmed at the genome level. This study focuses on the regulation of oxidative stress-related genes, as well as on genes that regulate the inflammatory response, apoptosis, response to heat stress, lipid metabolism, and the intestinal barrier status. The natural compounds presented include, but are not limited to, the following: rutin, lycopene, magnolol, genistein, hesperidin, naringin, quercetin, curcumin, bisdemethoxycurcumin, resveratrol, astaxanthin, squalene, pterostilbene, protocatechuic acid, taraxasterol, myricetin, and proanthocyanidins. Several studies have revealed a dose-dependent action. Future studies should focus on the role of phytogenic compounds as antibiotic alternatives in relation to gut microbiota and their role in eubiosis.
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Greece has a long tradition in cheesemaking, with 22 cheeses registered as protected designation of origin (PDO), 1 as protected geographical indication (PGI), and 1 applied for PGI. Several other cheeses are produced locally without any registration, which significantly contribute to the local economy. The present study investigated the composition (moisture, fat, salt, ash, and protein content), color parameters, and oxidative stability of cheeses that do not have a PDO/PGI certification, purchased from a Greek market. Milk and cheese types were correctly assigned for 62.8 and 82.1 % of samples, respectively, through discriminant analysis. The most important factors for milk type discrimination were L, a and b color attributes, salt, ash, fat-in-dry-matter, moisture-in-non-fat-substance, salt-in-moisture, and malondialdehyde contents, whereas a and b, and moisture, ash, fat, moisture-in-non-fat substance contents, and pH were the most influential characteristics for sample discrimination according to cheese type. A plausible explanation may be the differences in milk chemical composition between three animal species, namely cows, sheep, and goats and for the manufacture procedure and ripening. This is the very first report on the proximate analysis of these, largely ignored, chesses aiming to simulate interest for further study and production valorization.
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Fresh potatoes were deep-fried in olive oil (OO), extra virgin olive oil (EVOO), and their blends with 5%, 10%, and 20% v/v sesame oil (SO). This is the first report on the use of sesame oil as a natural source of antioxidants during olive oil deep frying. The oil was evaluated for anisidine value (AV), free fatty acids (FFAs), extinction coefficient (K232 and K270), Trolox equivalent antioxidant capacity (TEAC), and total phenols (TPs) until the total polar compounds (TPCs) reached 25%. Sesame lignan transformations were monitored through reversed-phase HPLC. While the TPCs in olive oils increased at a steady rate, the addition of 5%, 10%, and 20% v/v SO delayed TPCs' formation for 1, 2, and 3 h, respectively. The addition of 5%, 10%, and 20% v/v SO increased the olive oil frying time by 1.5 h, 3.5 h, and 2.5 h, respectively. The addition of SO to OO reduced the secondary oxidation products' formation rate. The AV for EVOO was lower than OO and all tested blends, even those with EVOO. EVOO was more resistant to oxidation than OO, as measured by the TPCs and TEAC, while the frying time rose from 21.5 to 25.25 h when EVOO replaced OO. The increase in frying time for OO but not for EVOO, after SO addition, points to a niche market for EVOO in deep frying.
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Aceites de Plantas , Aceite de Sésamo , Aceite de Oliva , Oxidación-Reducción , Antioxidantes/farmacologíaRESUMEN
Pomegranate is a fruit bearing-plant that is well known for its medicinal properties. Pomegranate is a good source of phenolic acids, tannins, and flavonoids. Pomegranate juice and by-products have attracted the scientific interest due to their potential health benefits. Currently, the medical community has showed great interest in exploiting pomegranate potential as a protective agent against several human diseases including cancer. This is demonstrated by the fact that there are more than 800 reports in the literature reporting pomegranate's anticancer properties. This review is an update on the research outcomes of pomegranate's potential against different types of human diseases, emphasizing on cancer. In addition, perspectives of potential applications of pomegranate, as a natural additive aiming to improve the quality of animal products, are discussed.
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Vinification by-products display great potential for utilization as feed additives rich in antioxidant compounds. Thus, the effect of dietary ground grape pomace (GGP), wine lees extract rich in yeast cell walls (WYC), and grape stem extracts (PE) on the relative expression of several genes involved in liver oxidative mechanisms and the oxidative status of the blood and breast muscle of broiler chickens was investigated. In total, 240 one-day-old as hatched chicks (Ross 308) were assigned to four treatments, with four replicate pens and 15 birds in each pen. Birds were fed either a basal diet (CON) or a basal diet supplemented with 25 g/kg GGP, or 2 g/kg WYC, or 1 g starch including 100 mg pure stem extract/kg (PE) for 42 days. The polyphenolic content of vinification by-products was determined using an LC-MS/MS library indicating as prevailing compounds procyanidin B1 and B2, gallic acid, caftaric acid, (+)-catechin, quercetin, and trans-resveratrol. Body weight and feed consumption were not significantly affected. The relative transcript level of GPX1 and SOD1 tended to increase in the liver of WYC-fed broilers, while NOX2 tended to decrease in the PE group. SOD activity in blood plasma was significantly increased in WYC and PE compared to the CON group. The total antioxidant capacity measured with FRAP assay showed significantly higher values in the breast muscle of PE-fed broilers, while the malondialdehyde concentration was significantly decreased in both WYC- and PE-fed broilers compared to the CON group. The exploitation of vinification by-products as feed additives appears to be a promising strategy to improve waste valorization and supply animals with bioactive molecules capable of improving animals' oxidative status and products' oxidative stability.
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The beneficial properties of the flavanones hesperidin and naringin as feed additives in poultry have lately been under investigation. In broilers, both flavanones have been shown to exhibit antioxidant properties while their individual effects on fatty acid (FA) composition and the underlying molecular mechanisms of their activity have not been explored. Here, we studied their effects on broiler meats' FA profiles and on the expression of genes related to lipid metabolism, antioxidant defense and anti-inflammatory function. The experimental design comprised six treatment groups of broilers, each supplemented from day 11 until slaughter at 42 days with hesperidin, naringin or vitamin E, as follows: the E1 group received 0.75 g of hesperidin per kg of feed, E2 received 1.5 g hesperidin/kg feed, N1 received 0.75 g naringin/kg feed, N2 received 1.5 g naringin/kg feed, vitamin E (VE) received 0.2 g a-tocopheryl acetate/kg feed, and the control group was not provided with a supplemented feed. The VE treatment group served as a positive control for antioxidant activity. An analysis of the FA profiles of the abdominal adipose tissue (fat pad), major pectoralis (breast) and biceps femoris (thigh) muscles showed that both hesperidin and naringin had significant effects on saturated FA (SFA), polyunsaturated FA (PUFA) and omega n-6 content. Both compounds reduced SFA and increased PUFA and n-6 content, as well as reducing the atherogenicity and thrombogenicity indices in the breast muscle and fat pad. The effects on the thigh muscle were limited. An analysis of gene expression in the liver revealed that naringin significantly increased peroxisome proliferator-activated receptor alpha (PPARα), Acyl-CoA oxidase 1 (ACOX1) and glutathione disulfide reductase (GSR) expression. In the breast muscle, both hesperidin and naringin increased fatty acid synthase (FASN) expression and hesperidin increased the expression of adiponectin. In brief, both hesperidin and naringin supplementation beneficially affected FA profiles in the breast meat and fat pad of broiler chicken. These effects could be attributed to an increase in FA ß-oxidation since the increased expression of related genes (PPARα and ACOX1) was observed in the liver. Furthermore, the antioxidant activity of hesperidin and naringin previously observed in the meat of broilers could be attributed, at least partly, to the regulation of antioxidant defense genes, as evidenced by the increased GSR expression in response to naringin supplementation.
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Mycotoxins appear to be the "Achilles' heel" of the agriculture sector inducing enormous economic losses and representing a severe risk to the health of humans and animals. Although novel determination protocols have been developed and legislation has been implemented within Europe, the side effects of mycotoxins on the homeostatic mechanisms of the animals have not been extensively considered. Feed mycotoxin contamination and the effects on the antioxidant status of livestock (poultry, swine, and ruminants) are presented. The findings support the idea that the antioxidant systems in both monogastrics and ruminants are challenged under the detrimental effect of mycotoxins by increasing the toxic lipid peroxidation by-product malondialdehyde (MDA) and inhibiting the activity of antioxidant defense mechanisms. The degree of oxidative stress is related to the duration of contamination, co-contamination, the synergetic effects, toxin levels, animal age, species, and productive stage. Since the damaging effects of MDA and other by-products derived by lipid peroxidation as well as reactive oxygen species have been extensively studied on human health, a more integrated monitoring mechanism (which will take into account the oxidative stability) is urgently required to be implemented in animal products.
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The objective of the present study was to investigate the effect of the natural flavonoid quercetin dietary supplementation on the alteration of egg metallome by applying the basic principles of elemental metabolomics. One hundred and ninety-two laying hens were allocated into 4 treatment groups: the control (C) group that was fed with a commercial basal diet and the other experimental groups that were offered the same diet further supplemented with quercetin at 200, 400 and 800 mg per kg of feed (Q2, Q4 and Q8 group, respectively) for 28 days. The diets contained the same vitamin and mineral premix, thus all birds received the same amount of elements since no differences on feed intake existed. The egg elemental profile consisted of As, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, Pb, Sb, Se, Sr, V, Zn and was determined using inductively coupled plasma mass spectrometry (ICP-MS). Quercetin supplementation altered the elemental profile. Most notably, quercetin altered the element concentrations predominantly in egg shell and albumen. It increased the concentration of Sb while reduced that of Cr and Se in both egg shell and albumen. Moreover, it increased As, Cd in albumen and V in yolk, while compared to the control, reduced As, Cd, Cr, Cu and V and also raised Ca, Fe, Mg and Ni in egg shell. The presence of quercetin led to differentiation of the deposition of certain trace minerals in egg compartments compared to that of hens fed a basal diet, possibly indicating that tailor made eggs for specific nutritional and health requirements could be created in the future.
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The basic principles of elemental metabolomics were applied to investigate whether alteration of egg metallome could be achieved after two flavonoids addition, namely hesperidin and naringin in diets of laying hens. A total of 72 hens were divided into six groups: Control (C) (basal diet), E1 (750 mg hesperidin/kg diet), E2 (1500 mg hesperidin/kg diet), N1 (750 mg naringin/kg diet), N2 (1500 mg naringin/kg diet), and VE (200 mg vitamin E/kg diet). The same diet was provided to birds of all treatments, with the exception of added supplements. The diets had the same vitamin and mineral premix; thus, all birds received the same number of elements because no differences on feed intake existed. The egg elemental profile consisted of As, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Mo, Ni, Pb, Sb, Se, Sr, V, Zn, and was determined using ICP-MS. Flavonoid supplementation altered the elemental profile. Most notably, in both albumen and yolk, hesperidin increased Ni, Pb, and Sr concentration while it decreased that of Co and Sb. Naringin increased Cd, Cr, Cu, Ni, and V and lowered the concentration of Co and Sb in both yolk and albumen. Vitamin E supplementation, in comparison to the control, decreased Co in both albumen and yolk and also raised Sb in albumen. Flavonoid presence led to the differences in deposition of certain trace minerals in egg compared to that of hens fed a basal diet or a diet with vitamin E supplementation.
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Selenium (Se) is an important trace mineral having many essential roles at the cellular and organismal levels in animal and human health. The biological effects of Se are mainly carried out by selenoproteins (encoded by 25 genes in humans and 24 in mice). As an essential component of selenoproteins, Se performs structural and enzymic roles; in the latter context it is well known for its catalytic and antioxidative functions. Studies involving different animal models have added great value to our understanding regarding the potential implications of Se and selenoproteins in mammalian fertility and reproduction. In this review, we highlight the implications of selenoproteins in male fertility and reproduction followed by the characteristic biological functions of Se and selenoproteins associated with overall male reproductive function. It is evident from observations of past studies (both animal and human) that Se is essentially required for spermatogenesis and male fertility, presumably because of its vital role in modulation of antioxidant defense mechanisms and other essential biological pathways and redox sensitive transcription factors. However, bearing in mind the evidences from mainstream literature, it is also advisable to perform more studies focusing on the elucidation of additional roles played by the peculiar and canonical selenoproteins i.e., glutathione peroxidase 4 (GPX4) and selenoprotein P (SELENOP) in the male reproductive functions. Nevertheless, search for the elucidation of additional putative mechanisms potentially modulated by other biologically relevant selenoproteins should also be included in the scope of future studies. However, as for the implication of Se in fertility and reproduction in men, though a few clinical trials explore the effects of Se supplementation on male fertility, due to inconsistencies in the recruitment of subjects and heterogeneity of designs, the comparison of such studies is still complicated and less clear. Therefore, further research focused on the roles of Se and selenoproteins is awaited for validating the evidences at hand and outlining any therapeutic schemes intended for improving male fertility. As such, new dimensions could be added to the subject of male fertility and Se supplementation.
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Selenium, through incorporation into selenoproteins, is one of the key elements of the antioxidant system. Over the past few years there has been increased interest in exploring those molecular mechanisms in chicken, responsible for the development of this protection system. In more detail, Cd/Pb poisoning and heat stress increase oxidation, mRNA levels of inflammatory proteins, and apoptotic proteins. Selenium seems to enhance the antioxidant status and alleviates these effects via upregulation of antioxidant proteins and other molecular effects. In this review, we analyze avian transcriptome key elements with particular emphasis on interactions with heavy metals and on relation to heat stress.
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The present work was part of a project intended to evaluate whether organic selenium (Se) has the potential to protect against toxic effects exerted by cadmium (Cd). For this reason, 300 as-hatched, one-day-old broiler chickens were randomly allocated in four dietary treatments with five replicate pens per treatment. Chickens in T1 treatment, were offered a diet supplemented with 0.3 ppm Se (as Se-yeast), without added Cd; in T2 treatment, they were offered a diet with 0.3 ppm Se and 10 ppm Cd; in T3 treatment, they were offered a diet with 0.3 ppm Se and 100 ppm Cd; in T4 treatment, chickens were offered a diet supplemented with 3 ppm Se and 100 ppm Cd. Cadmium was added to the diets in T2, T3, and T4 as CdCl2. On the fourth and sixth weeks, liver and breast samples were obtained from two broilers per replicate pen. Relative gene expression levels of catalase (CAT), superoxide dismutase 1 (SOD1) and 2 (SOD2), methionine sulfoxide reductase A (MSRA) and B3 (MSRB3), iodothyronine deiodinase 1 (DIO1), 2 (DIO2), and 3 (DIO3), glutathione peroxidase 1 (GPX1) and 4 (GPX4), thioredoxin reductase 1 (TXNRD1) and 3 (TXNRD3), and metallothionein 3 (MT3) were analyzed by real-time quantitative PCR in liver, whereas the fatty-acid (FA) profile of breast muscle was determined by gas chromatography. Broilers supplemented with 0.3 ppm Se could tolerate low levels of Cd present in the diets, as there were no significant changes in the breast muscle FA profile, whereas excess Cd led to decreased polyunsaturated fatty acids (PUFAs), and in particular n-6 PUFA. Furthermore, treatments mainly affected the messenger RNA (mRNA) expression of SOD2, TXNRD3, and MT3, while age affected CAT, MSRB3, DIO2, DIO3, GPX4, TXNRD1, and MT3. In conclusion, dietary Se may help against the negative effects of Cd, but cannot be effective when Cd is present at excessive amounts in the diet.
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Selenium (Se) is an essential trace element of fundamental importance to health due to its antioxidant, anti-inflammatory, and chemopreventive properties, attributed to its presence within at least 25 selenoproteins (Sel). In this review, we describe some of the recent progress, in our understanding, on the impact of maternal Se intake during the periconceptional period on offspring development and health. Maternal nutrition affects the performance and health of the progeny, and both maternal and offspring Se supplementations are essential for the optimal health and antioxidant protection of the offspring. The case of Se in epigenetic programming and early life nutrition is also discussed.
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This study presents the comprehensive elemental profile of Greek Graviera (Gruyère) cheeses. In total, 105 samples from nine different geographic regions produced from sheep, goat and cow milk and their mixtures were assessed. Elemental signatures of 61 elements were investigated for determination of geographic origin and milk type. Regional and milk type classification through Linear Discriminant Analysis was successful for almost all cases, while a less optimistic cross validation exercise presented lower classification rates. That points to further research using a much larger sample set, increasing confidence for cheese authentication utilizing also bioinformatics tools under development. This is the first study reporting signatures of 61 elements in dairy products including all sixteen rare earth elements and all seven precious metals. Safety and quality were assessed regarding toxic and nutritive elements. According to both EU and USA regulations and directives, Graviera is a nutritional source for trace and macro elements with low levels of toxic elements.
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Queso/análisis , Microbiología de Alimentos , Metabolómica , Animales , Bovinos , Queso/microbiología , Productos Lácteos/análisis , Femenino , Inocuidad de los Alimentos , Cabras , Grecia , Humanos , Leche/química , Leche/metabolismo , Evaluación Nutricional , OvinosRESUMEN
Selenium (Se) is an essential micronutrient that has several important functions in animal and human health. The biological functions of Se are carried out by selenoproteins (encoded by twenty-five genes in human and twenty-four in mice), which are reportedly present in all three domains of life. As a component of selenoproteins, Se has structural and enzymatic functions; in the latter context it is best recognized for its catalytic and antioxidant activities. In this review, we highlight the biological functions of Se and selenoproteins followed by an elaborated review of the relationship between Se and female reproductive function. Data pertaining to Se status and female fertility and reproduction are sparse, with most such studies focusing on the role of Se in pregnancy. Only recently has some light been shed on its potential role in ovarian physiology. The exact underlying molecular and biochemical mechanisms through which Se or selenoproteins modulate female reproduction are largely unknown; their role in human pregnancy and related complications is not yet sufficiently understood. Properly powered, randomized, controlled trials (intervention vs. control) in populations of relatively low Se status will be essential to clarify their role. In the meantime, studies elucidating the potential effect of Se supplementation and selenoproteins (i.e., GPX1, SELENOP, and SELENOS) in ovarian function and overall female reproductive efficiency would be of great value.
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Reproducción , Selenio/metabolismo , Selenoproteínas/metabolismo , Animales , Femenino , Humanos , Ovario/fisiología , EmbarazoRESUMEN
Unlike other essential trace elements that interact with proteins in the form of cofactors, selenium (Se) becomes co-translationally incorporated into the polypeptide chain as part of 21st naturally occurring amino acid, selenocysteine (Sec), encoded by the UGA codon. Any protein that includes Sec in its polypeptide chain is defined as selenoprotein. Members of the selenoproteins family exert various functions and their synthesis depends on specific cofactors and on dietary Se. The Se intake in productive animals such as chickens affect nutrient utilization, production performances, antioxidative status and responses of the immune system. Although several functions of selenoproteins are unknown, many disorders are related to alterations in selenoprotein expression or activity. Selenium insufficiency and polymorphisms or mutations in selenoproteins' genes and synthesis cofactors are involved in the pathophysiology of many diseases, including cardiovascular disorders, immune dysfunctions, cancer, muscle and bone disorders, endocrine functions and neurological disorders. Finally, heavy metal poisoning decreases mRNA levels of selenoproteins and increases mRNA levels of inflammatory factors, underlying the antagonistic effect of Se. This review is an update on Se dependent antioxidant enzymes, presenting the current state of the art and is focusing on results obtained mainly in chicken.
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BACKGROUND: Body size in sheep is an important indicator of productivity, growth and health as well as of environmental adaptation. It is a composite quantitative trait that has been studied with high-throughput genomic methods, i.e. genome-wide association studies (GWAS) in various mammalian species. Several genomic markers have been associated with body size traits and genes have been identified as causative candidates in humans, dog and cattle. A limited number of related GWAS have been performed in various sheep breeds and have identified genomic regions and candidate genes that partly account for body size variability. Here, we conducted a GWAS in Frizarta dairy sheep with phenotypic data from 10 body size measurements and genotypic data (from Illumina ovineSNP50 BeadChip) for 459 ewes. RESULTS: The 10 body size measurements were subjected to principal component analysis and three independent principal components (PC) were constructed, interpretable as width, height and length dimensions, respectively. The GWAS performed for each PC identified 11 significant SNPs, at the chromosome level, one on each of the chromosomes 3, 8, 9, 10, 11, 12, 19, 20, 23 and two on chromosome 25. Nine out of the 11 SNPs were located on previously identified quantitative trait loci for sheep meat, production or reproduction. One hundred and ninety-seven positional candidate genes within a 1-Mb distance from each significant SNP were found. A guilt-by-association-based (GBA) prioritization analysis (PA) was performed to identify the most plausible functional candidate genes. GBA-based PA identified 39 genes that were significantly associated with gene networks relevant to body size traits. Prioritized genes were identified in the vicinity of all significant SNPs except for those on chromosomes 10 and 12. The top five ranking genes were TP53, BMPR1A, PIK3R5, RPL26 and PRKDC. CONCLUSIONS: The results of this GWAS provide evidence for 39 causative candidate genes across nine chromosomal regions for body size traits, some of which are novel and some are previously identified candidates from other studies (e.g. TP53, NTN1 and ZNF521). GBA-based PA has proved to be a useful tool to identify genes with increased biological relevance but it is subjected to certain limitations.
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Tamaño Corporal/genética , Sitios de Carácter Cuantitativo , Ovinos/genética , Animales , Cromosomas/genética , Femenino , Estudio de Asociación del Genoma Completo , Polimorfismo de Nucleótido SimpleRESUMEN
Spatiotemporal release of growth factors from a delivery device can profoundly affect the efficacy of bone growth induction. Here, we report on a delivery platform based on the encapsulation of insulin-like growth factor I (IGF-I) in different poly(D,L-lactide) (PLA) and poly(D,L-lactide-co-glycolide) (PLGA) microsphere (MS) formulations to control IGF-I release kinetics. In vitro IGF-I release profiles generally exhibited an initial burst (14-36% of total IGF-I content), which was followed by a more or less pronounced dormant phase with little release (2 to 34 days), and finally, a third phase of re-increased IGF-I release. The osteoinductive potential of these different IGF-I PL(G)A MS formulations was tested in studies using 8-mm metaphyseal drill hole bone defects in sheep. Histomorphometric analysis at 3 and 6 weeks after surgery showed that new bone formation was improved in the defects locally treated with IGF-I PL(G)A MS (n=5) as compared to defects filled with IGF-I-free PL(G)A MS (n=4). The extent of new bone formation was affected by the particular release kinetics, although a definitive relationship was not evident. Local administration of IGF-I resulted in down-regulation of inflammatory marker genes in all IGF-I treated defects. The over-expression of growth factor genes in response to IGF-I delivery was restricted to formulations that produced osteogenic responses. These experiments demonstrate the osteoinductive potential of sustained IGF-I delivery and show the importance of delivery kinetics for successful IGF-I-based therapies.
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Regeneración Ósea/efectos de los fármacos , Huesos/efectos de los fármacos , Sistemas de Liberación de Medicamentos , Sustancias de Crecimiento/administración & dosificación , Factor I del Crecimiento Similar a la Insulina/administración & dosificación , Cicatrización de Heridas/efectos de los fármacos , Animales , Huesos/lesiones , Huesos/patología , Huesos/fisiología , Línea Celular Tumoral , Composición de Medicamentos , Implantes de Medicamentos , Regulación de la Expresión Génica/efectos de los fármacos , Sustancias de Crecimiento/química , Sustancias de Crecimiento/farmacología , Sustancias de Crecimiento/uso terapéutico , Humanos , Mediadores de Inflamación/antagonistas & inhibidores , Mediadores de Inflamación/metabolismo , Factor I del Crecimiento Similar a la Insulina/química , Factor I del Crecimiento Similar a la Insulina/farmacología , Factor I del Crecimiento Similar a la Insulina/uso terapéutico , Cinética , Ácido Láctico/química , Microesferas , Osteogénesis/efectos de los fármacos , Poliésteres/química , Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Distribución Aleatoria , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/química , Proteínas Recombinantes/farmacología , Proteínas Recombinantes/uso terapéutico , Oveja Doméstica , SolubilidadRESUMEN
Thyroid hormones increase cystatin C levels in vivo. To study whether 3,3',5-triiodo-l-thyronine (T(3)) stimulates the production of cystatin C in vitro, we used a T(3)-responsive osteoblastic cell line (PyMS) which can be kept in serum-free culture. We compared the effects of T(3) on cystatin C mRNA expression (by Northern) and on protein release (by Western and ELISA) with those of dexamethasone (dex). Triiodothyronine increased cystatin C mRNA expression and cystatin C accumulation in culture media in a dose- and time-dependent manner, 1.5-fold at 1 nmol/l after 4d; dex (100 nmol/l) was more potent and increased cystatin C accumulation 3-fold after 4d. Triiodothyronine but not dex stimulated glucose uptake. Our in vitro findings explain in vivo observations. Triiodothyronine-induced increase in the production of cystatin C may be related to an increased cell metabolism and proteolysis control demand.
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Cistatina C/biosíntesis , Osteoblastos/efectos de los fármacos , Triyodotironina/farmacología , Fosfatasa Alcalina/biosíntesis , Animales , Células Cultivadas , Desoxiglucosa/análogos & derivados , Desoxiglucosa/metabolismo , Dexametasona/farmacología , Transportador de Glucosa de Tipo 3/biosíntesis , Osteoblastos/metabolismo , ARN Mensajero/biosíntesis , RatasRESUMEN
Thyroid hormones increase energy expenditure and bone turnover in vivo. To study whether 3,3',5-triiodo-l-thyronine (T3) stimulates the uptake of glucose in osteoblastic cells, PyMS (a cell line derived from rat bone) cells were kept in serum-free culture medium and treated with T3. We measured [1-¹4C]-2-deoxy-D: -glucose (2DG) uptake and looked for expression of the high-affinity glucose transporters GLUT1 and GLUT3 by northern and western analysis. T3 did not influence the cell number but slightly (1.3-fold) increased the protein content of the cell cultures. 2DG uptake was low in serum-deprived cell cultures and was increased by T3 (up to 2.5-fold at 1 nmol l⻹ after 4 days) in a dose- and time-dependent manner. Triiodothyronine at 1 nmol l⻹ increased GLUT1 and GLUT3 abundance in membranes. Therefore, increased glucose uptake induced by T3 in osteoblasts may be mediated by the known high-affinity glucose transporters GLUT1 and GLUT3.