Effect of LPS and glutathione depletion on gene expression profiling in mouse macrophage-like cells
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE79397 | GEO | 2016/03/19
SECONDARY ACCESSION(S): PRJNA315698
REPOSITORIES: GEO
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Project description:Purpose: To better understand the effects of glutathione (GSH)-deficiency on lens homeostasis and cataractogenesis. Methods: The transcriptome of lens epithelia and fiber cells was obtained from C57BL/6 LEGSKO (lens GSH synthesis knockout) and buthionine sulfoximine (BSO)-treated LEGSKO mice and compared to C57BL/6 wild-type mice using RNA-Seq. Results: RNA-Seq results were in excellent agreement with qPCR (correlation coefficients between 0.87-0.94 and p<5E-6 for a subset of 36 mRNAs). Of the 24,415 transcripts mapped to the mouse genome, 441 genes showed significantly modulated expression. Pathway analysis indicated major changes in EMT signaling, visual cycle, small molecule biochemistry, and lipid metabolism. GSH-deficient lenses showed upregulation of genes relating to detoxification, including Aldh1a1, Aldh3a1 (aldehyde dehydrogenases), Mt1, Mt2 (metallothioneins), Ces1g (carboxylesterase), and Slc14a1 (urea transporter UT-B). These proteins share substrate specificity with GSH or glutathione-S-transferase and may protect GSH-deficient lenses. Genes associated with canonical EMT pathways, including Wnt10a, Egf, and Syk, showed upregulation in lens epithelia samples. Severely GSH-deficient lens epithelia showed a broad downregulation of vision-related genes (including Cryge, Crygf, and Rho). The BSO-treated LEGSKO lens epithelia transcriptome has significant correlation (r=0.63,P<0.005) to that of lens epithelia undergoing EMT. Conclusions: These results show that GSH depletion of the lens leads to expression of detoxifying genes and activation of EMT signaling, in addition to changes in transport systems and lipid homeostasis. These data give new insight into the adaptation and consequences of GSH-deficiency in the lens and suggest that supplementation of GSH or a precursor after cataract surgery could potentially reduce the incidence of EMT-mediated posterior subcapsular opacification.
2017-05-18 | GSE95688 | GEO
Project description:In order to understand how in vitro culture affects embryonic quality, we analyzed survival and global gene expression in bovine blastocysts after exposure to increased oxidative stress conditions of IVC. A pro-oxidant agents that act intra-cellularly by inhibiting GSH synthesis (0.4 mM buthionine sulfoximine [BSO]) was added from days 3 to 7, and transcriptomic analysis was then performed in resulting blastocysts. Precisely, after in vitro maturation and fertilization, bovine zygotes were culture in vitro in normal condition, then at day 3, embryos were allocated into culture in control or supplemented with BSO (0.4 mM) until day 7. At this time, blastocysts were harvested and analyzed. Our hypothesis was that BSO treatment will affect blastocyst survival and gene expression associated with low embryo quality
2012-11-14 | GSE42280 | GEO
Project description:In order to understand how in vitro culture affects embryonic quality, we analyzed survival and global gene expression in bovine blastocysts after exposure to increased oxidative stress conditions of IVC. A pro-oxidant agents that act intra-cellularly by inhibiting GSH synthesis (0.4 mM buthionine sulfoximine [BSO]) was added from days 3 to 7, and transcriptomic analysis was then performed in resulting blastocysts. Precisely, after in vitro maturation and fertilization, bovine zygotes were culture in vitro in normal condition, then at day 3, embryos were allocated into culture in control or supplemented with BSO (0.4 mM) until day 7. At this time, blastocysts were harvested and analyzed. Our hypothesis was that BSO treatment will affect blastocyst survival and gene expression associated with low embryo quality 4 replicates of 10 blast/rep was produced for BSO and control treatment. According to a dye swap design with 2 colors, 4 arrays were used to compare BSO (green) against CTL (red) and 4 arrays were used to compared BSO (red) againt CTL (green).
2012-11-14 | E-GEOD-42280 | biostudies-arrayexpress
Project description:Drug-induced liver injury (DILI) is a major problem in drug development and clinical drug therapy. Troglitazone (TGZ), a thiazolidinedione antidiabetic drug for the treatment of type II diabetes mellitus, was found to induce rare idiosyncratic severe liver injury in patients, which led to its withdrawal in 2000. In experimental animals in vivo, however, TGZ has never induced liver injury. In this study, administration of BALB/c female mice with TGZ alone significantly elevated alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels 6 h after the treatment. Co-administration of L-buthionine-(S,R)-sulfoximine (BSO), an inhibitor of glutathione synthesis, unexpectedly restrained the TGZ-dependent increase of ALT and AST. Furthermore, the ratio of oxidative stress marker glutathione/disulfide glutathione was significantly decreased in TGZ-alone- and BSO plus TGZ-administered mice, and increased hepatic mRNA levels of inflammation- and oxidative stress-related factors in liver were observed. Subsequently, microarray studies were performed 6 h after administration and we discovered that the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway was activated in TGZ-induced liver injury. Moreover, the activation of JAK/STAT pathway promoted phosphorylation of STAT3 in TGZ-alone- and BSO plus TGZ-administered mice. Consequently, upregulation of STAT3 activation increased mRNA levels of its downstream genes. In conclusion, the present study is the first time to establish a mouse model of TGZ-induced liver injury with BALB/c mouse for assessing DILI in drug development and the activation of JAK/STAT signaling pathway might be as an important role involved in hepatotoxic mechanisms of TGZ.
2021-07-14 | GSE117257 | GEO
Project description:To comprehensively investigate the effects of glutathione on the gene expression, the microarray analysis was performed in the glutathione-fed wild-type Arabidopsis thaliana. Wild-type Arabidopsis (ecotype Columbia-0) were fed with 1 mM oxidized glutathione (GSSG) and 2 mM reduced glutathione (GSH) for comparison at equal nitrogen equivalents. To examine the effects of glutathione other than nitrogen at equal nitrogen equivalents, plants were fed with 3 mM NH4NO3. Plants grown by water were used as a control.
2023-08-01 | GSE228403 | GEO
Project description:To provide preliminary insights into metabolic and lipidomic characteristics in radioresistant triple-negative breast cancer (TNBC) cells and suggest potential therapeutic targets, we performed a comprehensive metabolic and lipidomic profiling of radioresistant MDA-MB-231 (MDA-MB-231/RR) TNBC cells and their parental cells using gas chromatography-mass spectrometry and nano electrospray ionization-mass spectrometry, followed by multivariate statistical analysis. Buthionine sulfoximine (BSO) and radiation were co-treated to radioresistant TNBC cells. The level of glutathione (GSH) was significantly increased, and the levels of GSH synthesis-related metabolites, such as cysteine, glycine, and glutamine were also increased in MDA-MB-231/RR cells. In contrast, the level of lactic acid was significantly reduced. In addition, reactive oxygen species (ROS) level was decreased in MDA-MB-231/RR cells. In the lipidomic profiles of MDA-MB-231/RR cells, the levels of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were significantly increased, whereas those of most of the phosphatidylinositol species were significantly decreased. BSO sensitized MDA-MB-231/RR cells to radiotherapy, which resulted in decreased GSH level and increased ROS level and apoptosis. Radioresistant TNBC cells showed distinct metabolic and lipidomic characteristics compared to their parental cells. We suggested activated GSH, PC, and PE biosynthesis pathways as potential targets for treating radioresistant TNBC cells. Particularly, enhanced radiosensitivity was achieved by inhibition of GSH biosynthesis in MDA-MB-231/RR cells.
2021-09-22 | ST001920 | MetabolomicsWorkbench
Project description:This report provides data on ferroptosis induced proteomic response in immortalized mouse embryonic fibroblasts which are 4-OH-TAM-inducible Gpx4−/− (referred to as Pfa1 cells). Pfa1 cells are a valuable, widespread and well characterized model for ferroptosis research. We used L-Buthionine-sulfoximine (BSO) to induce ferroptosis and collected samples at 0h, 24h and 48 h after treatment. Untreated cells were used as a control. Pfa1 cells (kind gift from Marcus Conrad, Munich) were cultured in RPMI-1640 media (2 g/L glucose, 10% FBS, 2 mM Gibco GlutaMAX Supplement, 1% pen/strep) at 37C with 5% CO2 in a humidified incubator. Cells (300k) were seeded on Corning 100 mm tissue-culture treated culture dishes and incubated overnight. On the next day Pfa1 cells were treated with or without 30 µM BSO.
2023-04-12 | PXD041415 | Pride
Project description:Expression profiling of wild type E. coli K-12 strain MG1655 with exogenous glutathione (GSH) supplementation, sub-inhibitory ciprofloxacin concentration and GSH with inhibitory ciprofloxacin concentration. We used microarrays to detail the global gene expression program underlying ciprofloxacin exposure and glutathione mediated abrogation of ciprofloxacin's anti-bacterial action.
2018-01-29 | GSE93896 | GEO
Project description:In the current study, we used a mass spectrometry-based redox proteomics approach to test responses of the cysteine (Cys) proteome to selective disruption of the Trx- and GSH-dependent systems. Auranofin (ARF) was used to inhibit Trx reductase without detectable oxidation of the GSH/GSSG couple, and buthionine sulfoximine (BSO) was used to deplete GSH without detectable oxidation of Trx1. Results for 606 Cys-containing peptides (peptidyl Cys) showed that 36% were oxidized over 1.3-fold by ARF, while BSO-induced oxidation of peptidyl Cys was only 10%. Mean fold oxidation of these peptides was also higher by ARF than BSO treatment. Analysis of potential functional pathways showed that ARF oxidized peptides associated with glycolysis, cytoskeleton remodeling, translation and cell adhesion. Of 60 peptidyl Cys oxidized due to depletion of GSH, 41 were also oxidized by ARF and included proteins of translation and cell adhesion but not glycolysis or cytoskeletal remodeling. Studies to test functional correlates showed that pyruvate kinase activity and lactate levels were decreased with ARF but not BSO, confirming the effects on glycolysis-associated proteins are sensitive to oxidation by ARF. These data show that the Trx system regulates a broader range of proteins than GSH system, support distinct function of Trx and GSH in cellular redox control, and show for the first time in mammalian cells selective targeting peptidyl Cys and biological pathways due to deficient function of the Trx system. ICAT-labeled Cys peptides were analyzed by reverse-phase liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Peptide eluents were monitored in an MS survey scan followed by ten data-dependent MS/MS scans on an LTQ-Orbitrap ion trap mass spectrometer (Thermo Finnigan, San Jose, CA). The LTQ was used to acquire MS/MS spectra (2 m/z isolation width, 35% collision energy, 5,000 AGC target, 200 ms maximum ion time). The Orbitrap was used to collect MS scans (300-1600 m/z, 1,000,000 AGC target, 500 ms maximum ion time, resolution 30,000). All data were converted from .raw files to the .dta format using ExtractMS version 2.0 (Thermo Finnigan, San Jose, CA). The acquired MS/MS spectra were searched against a concatenated target-decoy human RefSeq (release 37 - September 2009 - 38108 target proteins) database of the National Center for Biotechnology Information using the SEQUEST Sorcerer algorithm (version 3.11, SAGE-N) searching parameters included partially tryptic restriction, parent ion mass tolerance (+/- 20 ppm), dynamic modifications of oxidized Met (+15.9949 Da), differential ICAT-modified Cys (+9.0302 Da) and static ICAT modification of Cys (+227.1270 Da). The peptides were classified by charge state and tryptic state (fully and partial) and first filtered by mass accuracy (10 ppm for high-resolution MS), and then dynamically by increasing XCorr and deltaCn values to reduce protein false discovery rate to less than 1%, according to the target-decoy strategy.
2019-11-12 | PXD000353 | Pride
Project description:Natural killer (NK) cell function is markedly impaired in acute leukemia patients, weakening their antitumor immune response; however, the specific mechanisms underlying these functional deficits remain incompletely understood. Here, we reveal the critical role of glutathione (GSH) metabolism in maintaining NK cell homeostasis and function. Compared with healthy individuals, NK cells from acute leukemia patients exhibit markedly reduced GSH levels. GSH depletion impairs NK cell cytotoxic activity, cytokine production, and proliferative capacity. Notably, supplementation with glutathione reduced ethyl ester (GSHEE) enhances the antileukemic activity of NK cells. Mechanistically, GSH supports optimal NK cell function by improving mitochondrial function and promoting oxidative phosphorylation (OXPHOS). In summary, this study identifies disrupted GSH metabolism as a driver of NK cell dysfunction in acute leukemia and suggests that GSH supplementation may enhance NK cell-mediated antileukemic effects, offering a potential new strategy for NK cell-based immunotherapy.
2024-11-17 | GSE281768 | GEO