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Status |
Public on Jul 13, 2018 |
Title |
Marginal selenium deficiency down-regulates inflammation-related genes in splenic leukocytes of the mouse |
Organism |
Mus musculus |
Experiment type |
Expression profiling by array
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Summary |
Moderate selenium deficiency may lead to an impaired capacity to cope with health challenges. Functional effects of suboptimal selenium intake are not fully known, and biomarkers for an insufficient selenium supply are inadequate. We therefore fed mice diets of moderately deficient or adequate selenium intake for 6 weeks. Changes in global gene expression were monitored by microarray analysis in splenic leukocytes. Genes for four selenoproteins, Sepw1, Gpx1, Selh and Sep15, were the most significantly down-regulated in moderate selenium deficiency, and this was confirmed by quantitative polymerase chain reaction (qPCR). Classification of significantly affected genes revealed that processes related to inflammation, heme biosynthesis, DNA replication and transcription, cell cycle and transport were affected by selenium restriction. Down-regulation by moderate selenium deficiency of specific genes involved in inflammation and heme biosynthesis was confirmed by qPCR. Myeloperoxidase and lysozyme activities were decreased in selenium-restricted leukocytes, providing evidence for functional consequences. Genes for 31 nuclear factor (NF)-κB targets were down-regulated in moderate selenium deficiency, indicating an impaired NF-κB signaling. Together, the observed changes point to a disturbance in inflammatory response. The selenoproteins found here to be sensitive to selenium intake in murine leukocytes might also be useful as biomarkers for a moderate selenium deficiency in humans.
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Overall design |
Male C57BL/6J mice (3–4 wk of age) from Charles River (Sulzfeld, Germany) were randomly assigned to the selenium-deficient or selenium-adequate group (12 mice per group) with free access to food and water. The selenium-adequate diet (Se-adeq) was produced by mixing selenomethionine (Acros, Geel, Belgium) into the selenium-deficient diet (Se-def; No. C1045 with 50% carbohydrates, 17% protein, 5% fat, 4% fibre, and mixture of micronutrients; Altromin, Lage, Germany) containing 0.086mg Se/kg (Riese et al., Endocrinology 2006) to yield a selenium content of 0.15 mg/kg corresponding to the dietary reference intake for mice. Diets were fed as powder for 6wk until mice were killed in the non-fasted state. Animals were anesthetized with isofluran and blood with heparinized capillaries by puncture of the retroorbital plexus. Anesthetized animals were killed by cervical dislocation. Spleens were removed aseptically, placed on a sterile microscope slide and crushed with the end of a 6-ml syringe plunger. Released cells were diluted in 5ml of cell culture medium (RPMI with 5% fetal calf serum; Gibco, Karlsruhe, Germany). Clumps were dispersed by drawing and expelling the suspension repeatedly through a 6-ml syringe with a 20-gauge needle. A 100-μmmesh was used to remove clumps and particles and to receive a single cell suspension. The mesh was rinsed with 5 ml RPMI. After centrifugation (5 min, 200g) and washing with 10 ml RPMI, erythrocytes were lysed for 5 min in 5 ml of ammonium chloride lysis buffer (0.15 mol/L NH4Cl, 10 mmol/L KHCO3, 0.1 mmol/L Na2EDTA, pH 7.4). Thereafter, 15 ml of balanced salt solution containing 0.2% bovine serum albumin was added, and the suspension was centrifuged for 5 min at 200g. The pellet was suspended in 1 ml phosphate-buffered saline (PBS) and transferred into a 2-ml tube. Thirty microliters of the suspension was smeared on microscope slides and dried on air for counting. The cells of the remaining suspension were pelleted (5 min, 200g) and washed with 1 ml PBS. The pellet was frozen at−80°C until RNA isolation. As determined with May–Grünwald–Giemsa staining, the leukocyte population was independent of the selenium status and on average was composed of 79% lymphocytes, 18% granulocytes and 2% monocytes.
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Contributor(s) |
Kipp A, Banning A, Brigelius-Flohe R, Keijer J, van Schothorst EM |
Citation(s) |
22137268 |
Submission date |
Jul 12, 2018 |
Last update date |
Jul 15, 2018 |
Contact name |
Evert M. van Schothorst |
E-mail(s) |
evert.vanschothorst@wur.nl
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Organization name |
Wageningen University
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Lab |
Human and Animal Physiology
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Street address |
De Elst 1
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City |
Wageningen |
ZIP/Postal code |
6708 WD |
Country |
Netherlands |
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Platforms (1) |
GPL7042 |
Agilent-012694 Whole Mouse Genome G4122A (Probe Name version) |
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Samples (24)
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This SubSeries is part of SuperSeries: |
GSE117027 |
Expression profiling of selenium deficiency in mouse colon and splenic leukocytes |
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Relations |
BioProject |
PRJNA480872 |
Supplementary file |
Size |
Download |
File type/resource |
GSE117026_RAW.tar |
92.1 Mb |
(http)(custom) |
TAR (of TXT) |
GSE117026_normalized_data.txt.gz |
1.6 Mb |
(ftp)(http) |
TXT |
Processed data are available on Series record |
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