Subcellular distribution of selenium and Se-containing proteins in human liver

Selenium is an essential trace element in many living organisms. In the present paper, the subcellular distribution of selenium and Se-containing proteins in human liver samples, which were obtained from normal subjects who had an accidental death, was investigated by differential centrifugation and column chromatography. Selenium was mainly enriched in nuclei, mitochondria and cytosol. Almost half of Se existed in the nuclei due to their large amount in liver and high Se concentration. 15-30% of Se was found in small compounds with Mr<2000 in the liver components separated by dialysis. The average abundance of Se in small molecular mass species of whole-liver was 23.6%, which suggested most of Se associated with biological macromolecules. Eight kinds of Se-containing proteins with molecular mass of 335+/-20, 249+/-15, 106+/-11, 84.6+/-5.8, 70. 5+/-5.4, 45.6+/-1.5, 14.8+/-2.6, 8.5+/-1.2 kDa were found in the subcellular fractions of human liver. Among them the 335, 84.6 and 8. 5 kDa proteins were individually present in one subcellular fraction, whereas the others coexisted in two, three or four subcellular fractions. The most abundant Se-containing proteins, 70.5 and 14.8 kDa, accounted for 33.6% and 48.5% in the whole-liver soluble Se-containing protein, respectively. The former was enriched in cytosol and the latter was mainly present in nuclei and mitochondria.     

Effect of sex and time of sampling on selenium and glutathione peroxidase activity in tissues of mature rats

Sprague-Dawley rats were used to investigate variations in measures of glutathione peroxidase (GSH-Px) and selenium (Se) concentration resulting from diurnal cycles and sex. Mature rats (equal numbers of males and females) were killed at 4 h intervals over a 48 h period (0200, 0600, 1000, 1800 and 2200 h each day). Selenium and GSH-Px were measured in plasma, erythrocytes, and liver and kidney cytosols. Selenium concentrations did not vary diurnally, but plasma GSH-Px activities were higher during the light than dark periods. Males had greater plasma GSH-Px activities and Se concentrations (42 EU and .45 mg/kg, respectively) than females (35 EU and .41 mg/kg respectively). GSH-Px activities were also higher in male kidney cytosols than females (117 and 76 EU, respectively). Selenium and GSH-Px activities, however, were lower in male liver cytosols (.48 mg/kg and 272 EU) than females (1.19 mg/kg and 795 EU, respectively). These data suggest that Se is distributed differently in male and female rats and the difference in Se distribution is accomplished by differences in GSH-Px activities.     

Subcellular distribution of selenium in deficient mouse liver.

Selenium (Se)-deficient mice were labelled in vivo with single pulses of [75Se]selenite, and the intrahepatic distribution of the trace element was studied by subcellular fractionation. At 1 h after intraperitoneal injection of 3.3 or 10 micrograms of Se/kg body weight, 15% of the respective doses were found in the liver. Accumulation in the subcellular fractions followed the order: Golgi vesicular much greater than lysosomal greater than cytosolic = microsomal greater than mitochondrial, peroxisomal, nuclear and plasma-membrane fraction. At a dose of 3.3 micrograms/kg, more than 90% of the hepatic Se was protein-bound. When cross-contamination was accounted for, the following specific Se contents of the subcellular compartments were extrapolated: Golgi apparatus, 7.50 pmol/mg; cytosol, 0.90 pmol/mg; endoplasmic reticulum, 0.80 pmol/mg; mitochondria, 0.49 pmol/mg; nuclei, lysosomes, peroxisomes and plasma membrane, less than 0.4 pmol/mg. At 10 micrograms/kg, a roughly 2-3-fold increase in Se content of all fractions was found without major changes in the intrahepatic distribution pattern. An extraordinary rise in the cytosolic fraction was due to an apparently non-protein-bound Se pool. At 24 h after dosing, total hepatic Se had decreased to 6% of the initial dose and had become predominantly protein-bound. The 60% decrease in hepatic Se was reflected in a similar fall in the subcellular levels of the trace element. The Golgi apparatus still had the highest specific Se content, although accumulation was 5 times less than that after 1 h. The cytosolic pool accounted for 50% of the hepatic Se at both labelling times. After 1 h the Golgi apparatus was, with 19%, the second largest intrahepatic pool, followed by the endoplasmic reticulum with 16%. The high affinity and fast response of the Golgi apparatus to Se supplementation of deficient mice is interpreted in terms of a predominant function of this cell compartment in the processing and the export of Se-proteins from the liver.     

Effect of fat on human liver selenium concentration

Liver samples from 94 adult men and eight fetuses were obtained at autopsy from the Helsinki region. After lyophilization, the samples were analyzed for selenium (Se) both before and after extraction of fat with hexane/isopropanol. An inverse exponential relationship (R ²=0.63) existed between unextracted adult liver Se concentration and the corresponding liver fat concentration. The relationship ceased to exist when Se was determined on the defatted tissue. The mean defatted adult liver Se concentration, 16.8 μM/kg dry wt, was significantly higher (p<0.001) than that of unextracted samples, 12.1 μM/kg. The Se concentration of fetal liver was not affected by the fat extraction. An age relationship (p<0.01) was found between subjects under 25 y and those over 40 y when the Se concentration was expressed per unextracted liver tissue but when expressed per defatted liver tissue the difference between age groups widened to comprise subjects over 60 y. Hepatic fat (mean Se concentration 2.7 μM/kg) thus constitutes a diluting factor. We conclude that the Se concentration of fatty human liver results in an underestimation of the selenium status of subjects who have an unphysiologically high liver fat concentration.     

Selenium in cereals: improving the efficiency of agronomic biofortification in the UK

Wheat, despite its relatively low selenium (Se) concentration in the UK, is still an important dietary Se source and its biofortification by use of Se fertiliser may be an efficient means to increase the relatively low Se status of the population. We need to know more about the fate of Se applied to the soil and how to ensure the efficiency of Se application, and the three studies reported in this issue of Plant and Soil are timely and informative. Selenium in soil, both globally and locally, is notoriously variable; however, the soils in these studies yielded wheat grain Se concentrations in the narrow range of 16–44 ng/g. The low plant Se levels reported here are not surprising, given that selenite is the dominant Se form in these soils. A regression equation (which used total and extractable Se and extractable S as variables) explained a high proportion of the variance in grain Se concentration. Sulphur application (a common practice on UK wheat growing soils) had variable effects on grain Se concentration, depending on soil S status, pH and possibly other factors. A fertiliser methodology study investigated ways to optimise Se application for the purpose of biofortification. It was calculated that an application of a modest 10 g Se/ha as selenate would increase the grain Se concentration of UK wheat from around 30 ng/g to 300 ng/g. The national Se fertiliser program in Finland shows that this increase would have a large effect on population Se status. However, Se recovery in grain at this application rate is only 14%, and it can be argued that large-scale agronomic biofortification of cereals with Se would be somewhat wasteful of a relatively scarce trace element. Selenium’s effects and interactions in soil, plants, animals and humans are complex and often surprising and will keep researchers busy well into the future.     

Application of sodium selenate to cowpea (Vigna unguiculata L.) increases shoot and grain Se partitioning with strong genotypic interactions

BackgorundCowpea is a crop widely used in developing countries due its rusticity. Besides its rich genotypic variability, most breeding programs do not explore its potential to improve elements uptake. Selenium (Se) is a scarce element in most soils, resulting in its deficiency being common in human diets. This study aimed to evaluate the interaction between biofortification with Se and genotypic variation in cowpea, on the concentrations of Se in roots, leaves + stem and grains.MethodsTwenty-nine cowpea genotypes were grown in a greenhouse in the absence (control) and presence of Se (12.5 μg Se kg⁻¹ soil) as sodium selenate, in fully randomized scheme. The plants were cultivated until grains harvest. The following variables were determined: roots dry weight (g), leaves + stems dry weight (g), grains dry weight (g), Se concentration (mg kg⁻¹) in roots, leaves + stems and grains, and Se partitioning to shoots and grains.ResultsSelenium application increased the Se concentration in roots, leaves + stems and grains in all genotypes. At least twofold variation in grain Se concentration was observed among genotypes. Selenium application did not impair biomass accumulation, including grain dry weight. Genotype “BRS Guariba” had the largest Se concentration in grains and leaves + stems. Genotype MNC04-795 F-158 had the largest partitioning of Se to shoots and grain, due to elevated dry weights of leaves + stems and grain, and high Se concentrations in these tissues.ConclusionThis information might be valuable in future breeding programs to select for genotypes with better abilities to accumulate Se in grain to reduce widespread human Se undernutrition.     

Lead effects in the chick during selenium deficiency

1. Growing chicks (Gallus domesticus) were fed a selenium-deficient diet supplemented with 0 or 2000 ppm lead (Pb) and 0 or 0.1 ppm selenium (Se). 2. Selenium addition stimulated growth at 0 but not at 2000 ppm Pb, while Pb depressed growth at both levels of Se. 3. Selenium addition stimulated Se-dependent glutathione peroxidase (GSH-Px) activity in liver, but Pb was without effect on GSH-Px activity. 4. Lead addition increased non-protein sulfhydryl (NPSH) concentrations in liver, kidney and thigh muscle. NPSH levels were not altered by Se. 5. The reported antagonism between Pb and Se does not appear to be mediated through effects on GSH-Px or NPSH metabolism.     

Seasonal fluctuations of selenium and sulfur accumulation in selenium hyperaccumulators and related nonaccumulators

Some plants hyperaccumulate selenium (Se) up to 1% of dry weight. This study was performed to obtain insight into whole-plant Se fluxes in hyperaccumulators. Selenium hyperaccumulators Astragalus bisulcatus and Stanleya pinnata were monitored over two growing seasons for seasonal fluctuations in concentrations of Se and the chemically similar element sulfur (S). The related nonhyperaccumulators Astragalus sericoleucus, Oxytropis sericea and Thlaspi montanum were included for comparison. In both hyperaccumulators leaf Se decreased from April to October, coinciding with Se hyperaccumulation in flowers and seeds. Root Se levels were lowest in summer. Selenium concentration decreased with leaf age in both hyperaccumulators. Leaf S levels peaked in summer in all plant species, as did Se levels in nonhyperaccumulators. Selenium and S levels tended to be negatively correlated in hyperaccumulators, and positively correlated in nonhyperaccumulators. These results suggest a specific flow of Se in hyperaccumulator plants over the growing season, from root to young leaves in spring, followed by remobilization from aging leaves to reproductive tissues in summer, and back to roots in the autumn.     

Effects of excess dietary selenite on lead toxicity in sheep

The hypothesis that excess dietary selenite ameliorates lead (Pb) toxicosis in domestic sheep was tested. Twenty 6–8-yr-old ewes fed alfalfa pellets were assigned to the following treatments: (1) control; (2) 9.8 mg Pb/kg body weight (b.w.)/d as PbCO₃; (3) 3 mg Se/animal/d as Na₂SeO₃·5H₂O; or (4) a combination of treatments 2 and 3. The gelatin-encapsulated salts were given orally. The study was terminated on d 104, by which time three animals in the Pb group and all five animals in the Pb+Se group had died. All remaining animals were slaughtered on d 104. Lead and Se concentrations were determined in six biweekly-collected blood samples and in soft tissues and bone. Sheep on the control and Se treatments had similar feed intakes, body weights, and tissue Pb levels. Those in the Pb+Se group had lower feed intake, but higher blood Pb values compared with the Pb group. Feeding either element increased (P<0.05) the concentration of that element in blood, kidney, liver, spleen, and bone. Muscle-Pb concentrations were not affected (P<0.05) by treatment. Selenium concentrations in kidney, liver, and muscle were greater (P<0.05), whereas those in heart were less (P<0.05) for the Pb+Se group than for the Se Group. Clinical signs associated with Pb toxicosis noted in other animals were not observed in the poisoned sheep in this study. Selenite did not protect sheep against Pb toxicity and likely served as a synergistic factor.     

人肝脏组织亚细胞组分中含硒蛋白的分离与测定

硒是生物必需的微量元素 ,具有重要的生物化学功能 ,与人类和动物的健康及疾病密切相关[1,2 ] .生物体内 80 %以上的硒是以与蛋白质结合的形式存在 ,所以目前大量的研究集中在硒蛋白的分离、鉴定和功能研究等方面[3 ] .从 1998年起 ,我们对正常人肝组织中硒和含硒蛋白的亚细胞分布作了一些探索性的工作 ,发现含硒蛋白在各亚细胞组分中的分布有显著的不同[4 ,5] .前期工作采用的凝胶过滤柱层析法研究含硒蛋白 ,该法分离量大 ,但分辨率较差 ,分子量相近的蛋白质难以区分[4 ,5] .本工作采用ＳＤＳ 聚丙烯酰胺凝胶电泳 (ＳＤＳ ＰＡＧＥ)分离人…     

Activation of superoxide dismutase in selenium-deficient mice infected with influenza virus.

Selenium (Se) deficiency is associated with decreased activities of Se-dependent antioxidant enzymes, glutathione peroxidase (GPx) and thioredoxin reductase (TR), and with changes in the cellular redox status. We have previously shown that host Se deficiency is responsible for increased virulence of influenza virus in mice due to changes in the viral genome. The present study examines the antioxidant defense systems in the lung and liver of Se-deficient and Se-adequate mice infected with influenza A/Bangkok/1/79. Results show that neither Se status nor infection changed glutathione (GSH) concentration in the lung. Hepatic GSH concentration was lower in Se-deficient mice, but increased significantly day 5 post infection. No significant differences due to Se status or influenza infection were found in catalase activities. As expected, Se deficiency was associated with significant decreases in GPx and TR activities in both lung and liver. GPx activity increased in the lungs and decreased in the liver of Se-adequate mice in response to infection. Both Se deficiency and influenza infection had profound effects on the activity of superoxide dismutase (SOD). The hepatic SOD activity was higher in Se-deficient than Se-adequate mice before infection. However, following influenza infection, hepatic SOD activity in Se-adequate mice gradually increased. Influenza infection was associated with a significant increase of SOD activity in the lungs of Se-deficient, but not Se-adequate mice. The maximum of SOD activity coincided with the peak of pathogenesis in infected lungs. These data suggest that SOD activation in the lung and liver may be a part of a compensatory response to Se deficiency and/or influenza infection. However, SOD activation that leads to increased production of H(2)O(2) may also contribute to pathogenesis and to influenza virus mutation in lungs of Se-deficient mice.     

Selenium biofortification of high-yielding winter wheat (Triticum aestivum L.) by liquid or granular Se fertilisation

Selenium (Se) is an essential trace element for humans and livestock. In the UK, human Se intake and status has declined since the 1980s. This is primarily due to the increased use of wheat (Triticum aestivum L.) grown in UK soils which are naturally low in Se. The aim of this study was to determine the potential for increasing grain Se concentration in a high-yielding UK wheat crop using fertilisers. The crop response of winter-wheat to Se fertilisation was determined under standard field conditions in two consecutive years at up to 10 sites. Selenium fertilisers were applied as high-volume drenches of sodium selenate solution, or as granular Se-containing products. Yield and harvest index were unaffected by Se fertilisation. Under all treatments, grain Se concentration increased by 16–26 ng Se g^?1 fresh weight (FW) per gram Se ha^?1 applied. An application of 10 g Se ha^?1 would thereby increase the Se concentration of most UK wheat grain 10-fold from current ambient levels and agronomic biofortification of UK-grown wheat is feasible. Total recovery (grain and straw) of applied Se was 20–35%. The fate of Se in the food-chain and in the soil must be determined in order to optimize the efficiency of this process.     

Changes in the antioxidant defence and in selenium concentration in tissues of vanadium exposed rats

Vanadium is an element whose role as a micronutrient for humans is not yet completely established, but which has been shown to possess hypoglycaemic properties in diabetes. In an earlier study, we showed that in STZ-diabetic rats, exposure to 1 mg V per day has no effect on glycaemia or on antioxidant status. When the exposure was raised to 3 mg V per day there was a hypoglycaemic effect, together with reduced Se in the tissues, which reduced antioxidant defences. The aim of the present study was to examine whether exposure to 1 mg V per day modifies Se nutritional status and/or antioxidant defences in healthy rats. Two groups of rats were examined: control and vanadium-treated. Vanadium, as bis(maltolato)oxovanadium(iv), was supplied in the drinking water. The experiment had a duration of five weeks. Selenium was measured in excreta, serum, skeletal muscle, kidneys, liver, heart, femur and adipose tissue. Number of red (RBC) and white (WBC) blood cells and haemoglobin (Hb) were determined in samples of whole blood. Glutathione peroxidase (GPx), glutathione transferase (GST), catalase (CAT) and NAD(P)H:quinine-oxidoreductase1 (NQO1) activity, and malondialdehyde (MDA) in the liver were evaluated. Treatment significantly reduced food intake, produced an anaemic state, and decreased Se absorption and Se content in serum, kidneys and the liver. GPx, GST and NQO1 activity were decreased in the liver, while MDA levels rose. We conclude that healthy rats are more sensitive than diabetic ones to the effects of V. This should be taken into account for populations that are particularly exposed to V for environmental reasons, and/or that consume V as a nutritional supplement.     

Mercury and selenium interaction in vivo: effects on thioredoxin reductase and glutathione peroxidase

Mercury compounds exert toxic effects via interaction with many vital enzymes involved in antioxidant regulation, such as selenoenzymes thioredoxin reductase (TrxR) and glutathione peroxidase (GPx). Selenium supplementation can reactivate the mercury-inhibited TrxR and recover the cell viability in vitro. To gain an insight on how selenium supplementation affects mercury toxicity in vertebrates, we investigated the effects of selenium on the mercury accumulation and TrxR and GPx activities in a fish model. Juvenile zebra-seabreams were exposed either to methylmercury (MeHg) or inorganic mercury (Hg(2+)) in the presence or absence of sodium selenite (Se) for 28 days followed by 14 days of depuration. Mercury accumulation was found to be 10-fold higher under MeHg exposure than under Hg(2+) exposure. Selenium supplementation caused a half decrease of the accumulation of MeHg but did not influence Hg(2+) accumulation. Exposure to both mercurials led to a decrease of the activity of TrxR (<50% of control) in all organs. Se supplementation coincident with Hg(2+) exposure protected the thioredoxin system in fish liver. However, supplementation of Se during the depuration phase had no effects. The activity of GPx was only affected in the brain of fishes upon the exposure to MeHg and coexposure to MeHg and Se. Selenium supplementation has a limited capacity to prevent mercury effects in brain and kidney. These results demonstrate that Se supplementation plays a protective role in a tissue-specific manner and also highlight the importance of TrxR as a main target for mercurials in vivo.     

人体硒代谢与硒营养研究进展

硒是人体所必需的重要微量营养元素,综述了当前国内外人体硒代谢与硒营养的研究进展,包括硒源形式与吸收、人体的硒含量与分布、硒的代谢途径、硒的生物活化形式、硒与疾病、硒中毒和硒的安全摄入量。在此基础上,提出了针对我国硒资源分布、硒反应症分布和居民膳食结构硒摄入量的研究建议,为满足居民通过膳食和补充剂补硒预防和治疗疾病提供理论和实践指导。     

环境中硒的生物地球化学循环和营养调控及分异成因

硒是环境中重要的生命元素,它在环境中含量水平的高低直接影响着人及动植物的健康安全。结合国内外资料及最新的研究进展,阐述了环境中硒的生物地球球化学循环,包括环境中硒的生物地球化学循环特征,土壤中硒的含量分布、形态及有效性,大气和水环境中硒的形态分布,植物体中的硒及其对硒的吸收关系;讨论了低硒高硒环境中硒营养水平的调节及环境分异的成因,诸如母质类型、气候特征、风化淋失、气体挥发、土壤质地和地力耗竭等方面;并提出了环境中硒研究的前沿及今后关注的热点问题,以促进今后环境中硒的研究。     

Effect of dietary fluoride on selenite toxicity in the rat

Three factorial experiments were conducted to determine if high dietary fluoride (F) would inhibit selenite toxicity in rats. Initially, three levels of selenite (0.05, 3, and 5 mg/kg diet) were matched against three levels of F (2, 75, and 150 mg/kg diet). Fluoride failed to prevent the depressive effect of selenite on 8-wk food intake and body wt gain. Selenium (Se) concentration of plasma and kidney and enzymatic activity of whole blood glutathione peroxidase (GSH-Px) were also unaffected by F. Liver Se concentration, however, was slightly (12%) but significantly (p<0.025) reduced when the highest F and Se levels were combined. Fluoride (150 mg/kg) appeared to reduce liver selenite toxicity (5 mg/kg). Therefore, further study focused on liver histology with treatments that eliminated the middle levels of selenite and F. Fluoride prevented the hepatic necrosis seen in selenite-toxic rats. Similar histological lesions were not observed for kidney or heart. Fluoride partially (26%) but significantly (p<0.025) reduced thiobarbituric-reactive substances in selenite-toxic rats, but there was no F effect on intracellular distribution of liver Se, glutathione levels in liver and kidney, or on liver xanthine oxidase activity. Overall, the protective effect of F on selenite toxicity appears to be confined to liver pathology. The exact mechanism for this effect, however, remains unclear. Oregon Agricultural Experiment Station Technical Paper No. 9728.     

植物硒生理及与重金属交互的研究进展

硒是一种重要的微量元素,在低浓度时对生物有益,但高浓度时呈现与重金属类似的毒性。植物作为人体硒摄入的主要来源,其硒代谢对于植物硒积累乃至人体硒营养水平十分重要。研究植物硒吸收、代谢和积累机理能指导富硒粮食的生产,是解决人体硒摄入不足/超量问题的有效途径。本文在阐述土壤硒含量、形态、生物有效性及分布的基础上,综述了植物对硒的吸收、代谢机理的研究进展,并讨论了农业生物强化以及遗传育种生物强化等两种硒生物强化的实践方法,以及利用硒生物强化缓解重金属毒性减少积累;最后,提出了植物硒代谢及硒生物强化研究的前沿问题,以期为改善人体硒营养水平提高人体健康状况提供理论和实践依据。