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Scientific Reports Apr 20222-Deoxy-D-glucose (2DG) has recently received emergency approval for the treatment of COVID-19 in India, after a successful clinical trial. SARS-CoV-2 infection of...
2-Deoxy-D-glucose (2DG) has recently received emergency approval for the treatment of COVID-19 in India, after a successful clinical trial. SARS-CoV-2 infection of cultured cells is accompanied by elevated glycolysis and decreased mitochondrial function, whereas 2DG represses glycolysis and stimulates respiration, and restricts viral replication. While 2DG has pleiotropic effects on cell metabolism in cultured cells it is not known which of these manifests in vivo. On the other hand, it is known that 2DG given continuously can have severe detrimental effects on the rodent heart. Here, we show that the principal effect of an extended, intermittent 2DG treatment on mice is to augment the mitochondrial respiratory chain proteome in the heart; importantly, this occurs without vacuolization, hypertrophy or fibrosis. The increase in the heart respiratory chain proteome suggests an increase in mitochondrial oxidative capacity, which could compensate for the energy deficit caused by the inhibition of glycolysis. Thus, 2DG in the murine heart appears to induce a metabolic configuration that is the opposite of SARS-CoV-2 infected cells, which could explain the compound's ability to restrict the propagation of the virus to the benefit of patients with COVID-19 disease.
Topics: Animals; Deoxyglucose; Electron Transport; Glucose; Humans; Mice; Proteome; SARS-CoV-2; COVID-19 Drug Treatment
PubMed: 35478201
DOI: 10.1038/s41598-022-10168-1 -
Cells Dec 2020Bladder cancer is one of the most prevalent deadly diseases worldwide. Grade 2 tumors represent a good window of therapeutic intervention, whose optimization requires...
Bladder cancer is one of the most prevalent deadly diseases worldwide. Grade 2 tumors represent a good window of therapeutic intervention, whose optimization requires high resolution biomarker identification. Here we characterize energy metabolism and cellular properties associated with spreading and tumor progression of RT112 and 5637, two Grade 2 cancer cell lines derived from human bladder, representative of luminal-like and basal-like tumors, respectively. The two cell lines have similar proliferation rates, but only 5637 cells show efficient lateral migration. In contrast, RT112 cells are more prone to form spheroids. RT112 cells produce more ATP by glycolysis and OXPHOS, present overall higher metabolic plasticity and are less sensitive than 5637 to nutritional perturbation of cell proliferation and migration induced by treatment with 2-deoxyglucose and metformin. On the contrary, spheroid formation is less sensitive to metabolic perturbations in 5637 than RT112 cells. The ability of metformin to reduce, although with different efficiency, cell proliferation, sphere formation and migration in both cell lines, suggests that OXPHOS targeting could be an effective strategy to reduce the invasiveness of Grade 2 bladder cancer cells.
Topics: Cell Line, Tumor; Cell Movement; Cell Proliferation; Deoxyglucose; Energy Metabolism; Glycolysis; Humans; Microscopy, Confocal; Mitochondria; Neoplasm Grading; Oxidative Stress; Urinary Bladder Neoplasms
PubMed: 33322565
DOI: 10.3390/cells9122669 -
Neurochemical Research Jan 20172-Deoxy-D-[C]glucose ([C]DG) is commonly used to determine local glucose utilization rates (CMR) in living brain and to estimate CMR in cultured brain cells as rates of...
Determination of Glucose Utilization Rates in Cultured Astrocytes and Neurons with [C]deoxyglucose: Progress, Pitfalls, and Discovery of Intracellular Glucose Compartmentation.
2-Deoxy-D-[C]glucose ([C]DG) is commonly used to determine local glucose utilization rates (CMR) in living brain and to estimate CMR in cultured brain cells as rates of [C]DG phosphorylation. Phosphorylation rates of [C]DG and its metabolizable fluorescent analog, 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose (2-NBDG), however, do not take into account differences in the kinetics of transport and metabolism of [C]DG or 2-NBDG and glucose in neuronal and astrocytic cells in cultures or in single cells in brain tissue, and conclusions drawn from these data may, therefore, not be correct. As a first step toward the goal of quantitative determination of CMR in astrocytes and neurons in cultures, the steady-state intracellular-to-extracellular concentration ratios (distribution spaces) for glucose and [C]DG were determined in cultured striatal neurons and astrocytes as functions of extracellular glucose concentration. Unexpectedly, the glucose distribution spaces rose during extreme hypoglycemia, exceeding 1.0 in astrocytes, whereas the [C]DG distribution space fell at the lowest glucose levels. Calculated CMR was greatly overestimated in hypoglycemic and normoglycemic cells because the intracellular glucose concentrations were too high. Determination of the distribution space for [C]glucose revealed compartmentation of intracellular glucose in astrocytes, and probably, also in neurons. A smaller metabolic pool is readily accessible to hexokinase and communicates with extracellular glucose, whereas the larger pool is sequestered from hexokinase activity. A new experimental approach using double-labeled assays with DG and glucose is suggested to avoid the limitations imposed by glucose compartmentation on metabolic assays.
Topics: Animals; Astrocytes; Biological Transport; Brain; Carbon Radioisotopes; Cells, Cultured; Deoxyglucose; Dose-Response Relationship, Drug; Female; Glucose; Intracellular Fluid; Neurons; Pregnancy; Rats; Rats, Sprague-Dawley
PubMed: 26141225
DOI: 10.1007/s11064-015-1650-x -
Biochemical and Biophysical Research... Jan 2024Peritoneal dialysis (PD) and prolonged exposure to PD fluids (PDF) induce peritoneal membrane (PM) fibrosis and hypervascularity, leading to functional PM degeneration....
Peritoneal dialysis (PD) and prolonged exposure to PD fluids (PDF) induce peritoneal membrane (PM) fibrosis and hypervascularity, leading to functional PM degeneration. 2-deoxy-glucose (2-DG) has shown potential as PM antifibrotic by inhibiting hyper-glycolysis induced mesothelial-to-mesenchymal transition (MMT). We investigated whether administration of 2-DG with several PDF affects the permeability of mesothelial and endothelial barrier of the PM. The antifibrotic effect of 2-DG was confirmed by the gel contraction assay with embedded mesothelial (MeT-5A) or endothelial (EA.hy926) cells cultured in Dianeal® 2.5 % (CPDF), BicaVera® 2.3 % (BPDF), Balance® 2.3 % (LPDF) with/without 2-DG addition (0.2 mM), and qPCR for αSMA, CDH2 genes. Moreover, 2-DG effect was tested on the permeability of monolayers of mesothelial and endothelial cells by monitoring the transmembrane resistance (R), FITC-dextran (10, 70 kDa) diffusion and mRNA expression levels of CLDN-1 to -5, ZO1, SGLT1, and SGLT2 genes. Contractility of MeT-5A cells in CPDF/2-DG was decreased, accompanied by αSMA (0.17 ± 0.03) and CDH2 (2.92 ± 0.29) gene expression fold changes. Changes in αSMA, CDH2 were found in EA.hy926 cells, though αSMA also decreased under LPDF/2-DG incubation (0.42 ± 0.02). Overall, 2-DG mitigated the PDF-induced alterations in mesothelial and endothelial barrier function as shown by R, dextran transport and expression levels of the CLDN-1 to -5, ZO1, and SGLT2. Thus, supplementation of PDF with 2-DG not only reduces MMT but also improves functional permeability characteristics of the PM mesothelial and endothelial barrier.
Topics: Humans; Sodium-Glucose Transporter 2; Deoxyglucose; Endothelial Cells; Peritoneal Dialysis; Peritoneum; Dialysis Solutions; Peritoneal Fibrosis; Glucose; Epithelial Cells; Cells, Cultured
PubMed: 38104523
DOI: 10.1016/j.bbrc.2023.149376 -
Ecotoxicology and Environmental Safety Jan 2024Inhaling silica causes the occupational illness silicosis, which mostly results in the gradual fibrosis of lung tissue. Previous research has demonstrated that...
Inhaling silica causes the occupational illness silicosis, which mostly results in the gradual fibrosis of lung tissue. Previous research has demonstrated that hypoxia-inducible factor-1α (HIF-1α) and glycolysis-related genes are up-regulated in silicosis. The role of 2-deoxy-D-glucose (2-DG) as an inhibitor of glycolysis in silicosis mouse models and its molecular mechanisms remain unclear. Therefore, we used 2-DG to observe its effect on pulmonary inflammation and fibrosis in a silicosis mouse model. Furthermore, in vitro cell experiments were conducted to explore the specific mechanisms of HIF-1α. Our study found that 2-DG down-regulated HIF-1α levels in alveolar macrophages induced by silica exposure and reduced the interleukin-1β (IL-1β) level in pulmonary inflammation. Additionally, 2-DG reduced silica-induced pulmonary fibrosis. From these findings, we hypothesize that 2-DG reduced glucose transporter 1 (GLUT1) expression by inhibiting glycolysis, which inhibits the expression of HIF-1α and ultimately reduces transcription of the inflammatory cytokine, IL-1β, thus alleviating lung damage. Therefore, we elucidated the important regulatory role of HIF-1α in an experimental silicosis model and the potential defense mechanisms of 2-DG. These results provide a possible effective strategy for 2-DG in the treatment of silicosis.
Topics: Animals; Mice; Deoxyglucose; Glucose; Hypoxia-Inducible Factor 1, alpha Subunit; Inflammation; Macrophages, Alveolar; Pneumonia; Pulmonary Fibrosis; Silicon Dioxide; Silicosis
PubMed: 38039851
DOI: 10.1016/j.ecoenv.2023.115767 -
Cerebral Cortex (New York, N.Y. : 1991) Oct 2017In an attempt to shed light on the role of the prefrontal cortex in action perception, we used the quantitative 14C-deoxyglucose method to reveal the effects elicited by...
In an attempt to shed light on the role of the prefrontal cortex in action perception, we used the quantitative 14C-deoxyglucose method to reveal the effects elicited by reaching-to-grasp in the light or in the dark and by observation of the same action executed by an external agent. We analyzed the cortical areas in the principal sulcus, the superior and inferior lateral prefrontal convexities and the orbitofrontal cortex of monkeys. We found that execution in the light and observation activated in common most of the lateral prefrontal and orbitofrontal cortical areas, with the exception of 9/46-dorsal activated exclusively for observation and 9/46-ventral, 11 and 13 activated only for execution. Execution in the dark implicated only the ventral bank of the principal sulcus and its adjacent inferior convexity along with areas 47/12-dorsal and 13, whereas execution in the light activated both banks of the principal sulcus and both superior and inferior convexities along with areas 10 and 11. Our results demonstrate that the prefrontal cortex integrates information in the service of both action generation and action perception, and are discussed in relation to its contribution in movement suppression during action observation and in attribution of action to the correct agent.
Topics: Animals; Brain Mapping; Deoxyglucose; Female; Macaca mulatta; Movement; Perception; Prefrontal Cortex; Psychomotor Performance
PubMed: 27600843
DOI: 10.1093/cercor/bhw261 -
Nephrology, Dialysis, Transplantation :... Aug 2015Protein-bound uraemic toxins (PBUT), dicarbonyl stress and advanced glycation end products (AGEs) associate with cardiovascular disease in dialysis. Intensive...
BACKGROUND
Protein-bound uraemic toxins (PBUT), dicarbonyl stress and advanced glycation end products (AGEs) associate with cardiovascular disease in dialysis. Intensive haemodialysis (HD) may have significant clinical benefits. The aim of this study was to evaluate the acute effects of conventional and extended HD and haemodiafiltration (HDF) on reduction ratio (RR) and total solute removal (TSR) of PBUT, dicarbonyl stress compounds and AGEs.
METHODS
Thirteen stable conventional HD patients randomly completed a single study of 4-h HD (HD4), 4-h HDF (HDF4), 8-h HD (HD8) and 8-h HDF (HDF8) with a 2-week interval between the study sessions. RR and TSR of PBUT [indoxyl sulphate (IS), p-cresyl sulphate (PCS), p-cresyl glucuronide, 3-carboxyl-4-methyl-5-propyl-2-furanpropionic acid (CMPF), indole-3-acetic acid (IAA) and hippuric acid] of free and protein-bound AGEs [N(ε)-(carboxymethyl)lysine (CML), N(ε)-(carboxyethyl)lysine (CEL), Nδ-(5-hydro-5-methyl-4-imidazolon-2-yl)-ornithine, pentosidine], as well as of dicarbonyl compounds [glyoxal, methylglyoxal, 3-deoxyglucosone], were determined.
RESULTS
Compared with HD4, HDF4 resulted in increased RR of total and/or free fractions of IAA and IS as well as increased RR of free CML and CEL. HD8 and HDF8 showed a further increase in TSR and RR of PBUT (except CMPF), as well as of dicarbonyl stress and free AGEs compared with HD4 and HDF4. Compared with HD8, HDF8 only significantly increased RR of total and free IAA and free PCS, as well as RR of free CEL.
CONCLUSIONS
Dialysis time extension (HD8 and HDF8) optimized TSR and RR of PBUT, dicarbonyl stress and AGEs, whereas HDF8 was superior to HD8 for only a few compounds.
Topics: Blood Proteins; Deoxyglucose; Female; Glycation End Products, Advanced; Hemodiafiltration; Humans; Male; Middle Aged; Oxidative Stress; Renal Dialysis; Toxins, Biological; Uremia
PubMed: 25862762
DOI: 10.1093/ndt/gfv038 -
Zhongguo Ying Yong Sheng Li Xue Za Zhi... Sep 2022To investigate the effects of blocking lactate synthesis on the HT22 cell injuries caused by hypoxia.
OBJECTIVE
To investigate the effects of blocking lactate synthesis on the HT22 cell injuries caused by hypoxia.
METHODS
2-deoxy-D-glucose (2-DG) is a non-metabolized glucose analogue that can inhibit lactate synthesis by blocking glycolysis. HT22 cells were divided into 4 groups: Control group, 2-DG group, Hypoxia group and 2-DG+Hypoxia group. The cells in control group and 2-DG treatment group were cultured in a 37℃, 5% CO incubator, and thecells in hypoxia group and 2-DG + Hypoxia group were cultured in a hypoxia incubator. The concentrations of 2-DG were 2.5 and 5 mmol/L, the concentration of oxygen was 0.3%, and the treatment time was 24 h. Cell activity was detected by CCK-8 assay, the levels of lactate in cell culture medium were detected by spectrophotometry, cell morphology was observed by fluorescence staining, the level of reactive oxygen species (ROS) was detected by flow cytometry, and the activities of superoxide dismutase (SOD) and catalase (CAT) were determined by enzyme activity kits. The protein expression levels of p-p38, t-p38 and β-actin were detected by Western blot.
RESULTS
Compared with that in control group, the lactate level in culture medium and cell activity were decreased significantly (<0.01), the number of adherent cells was decreased, the level of ROS was increased (<0.01), and the enzyme activity of CAT was decreased (<0.05) in the 2-DG group. In the hypoxia group, the level of lactate in the culture medium was increased significantly (<0.01), the cell activity was decreased (<0.01), the number of adherent cells was decreased, the ROS levels were increased (<0.01), and the enzyme activities of CAT and SOD were decreased (<0.01 or <0.05). In 2-DG+Hypoxia group, the level of lactate was decreased significantly (<0.05), the cell viability was decreased significantly (<0.01), the number of cells was decreased significantly, and the ability of adhere to the wall was weakened significantly. The level of ROS was increased significantly (<0.01), the enzyme activities of CAT and SOD were decreased significantly (<0.01), the protein expression level of p-p38 was increased significantly (<0.05), and there was no change in t-p38. Compared with hypoxia groups, in 2-DG combined with hypoxia group, the level of lactate induced by hypoxia, the cell activity, and the enzyme activity level of CAT were decreased significantly (all <0.01), while the level of ROS was increased significantly (< 0.01).
CONCLUSION
Blocking lactate can reduce the cell activity level under hypoxia and aggravate the oxidative stress injury of HT22 cells. The mechanisms may be related to increasing ROS level and activating p38 signal pathway.
Topics: Humans; Reactive Oxygen Species; Lactic Acid; Hypoxia; Oxidative Stress; Neurons; Superoxide Dismutase; Deoxyglucose; Apoptosis
PubMed: 37088741
DOI: 10.12047/j.cjap.6276.2022.075 -
Metabolism: Clinical and Experimental Mar 2021GDI1 gene encodes for αGDI, a protein controlling the cycling of small GTPases, reputed to orchestrate vesicle trafficking. Mutations in human GDI1 are responsible for...
OBJECTIVES
GDI1 gene encodes for αGDI, a protein controlling the cycling of small GTPases, reputed to orchestrate vesicle trafficking. Mutations in human GDI1 are responsible for intellectual disability (ID). In mice with ablated Gdi1, a model of ID, impaired working and associative short-term memory was recorded. This cognitive phenotype worsens if the deletion of αGDI expression is restricted to neurons. However, whether astrocytes, key homeostasis providing neuroglial cells, supporting neurons via aerobic glycolysis, contribute to this cognitive impairment is unclear.
METHODS
We carried out proteomic analysis and monitored [F]-fluoro-2-deoxy-d-glucose uptake into brain slices of Gdi1 knockout and wild type control mice. d-Glucose utilization at single astrocyte level was measured by the Förster Resonance Energy Transfer (FRET)-based measurements of cytosolic cyclic AMP, d-glucose and L-lactate, evoked by agonists selective for noradrenaline and L-lactate receptors. To test the role of astrocyte-resident processes in disease phenotype, we generated an inducible Gdi1 knockout mouse carrying the Gdi1 deletion only in adult astrocytes and conducted behavioural tests.
RESULTS
Proteomic analysis revealed significant changes in astrocyte-resident glycolytic enzymes. Imaging [F]-fluoro-2-deoxy-d-glucose revealed an increased d-glucose uptake in Gdi1 knockout tissue versus wild type control mice, consistent with the facilitated d-glucose uptake determined by FRET measurements. In mice with Gdi1 deletion restricted to astrocytes, a selective and significant impairment in working memory was recorded, which was rescued by inhibiting glycolysis by 2-deoxy-d-glucose injection.
CONCLUSIONS
These results reveal a new astrocyte-based mechanism in neurodevelopmental disorders and open a novel therapeutic opportunity of targeting aerobic glycolysis, advocating a change in clinical practice.
Topics: Animals; Brain; Cells, Cultured; Deoxyglucose; Down-Regulation; Glucose; Glycolysis; Guanine Nucleotide Dissociation Inhibitors; Intellectual Disability; Male; Maze Learning; Memory; Memory Disorders; Mice; Mice, Knockout
PubMed: 33309713
DOI: 10.1016/j.metabol.2020.154463 -
Journal of Chromatographic Science Nov 2017The measurement of α-dicarbonyls and other degradation products of sugars has become important in view of their toxicity. Although there are several methods used for...
The measurement of α-dicarbonyls and other degradation products of sugars has become important in view of their toxicity. Although there are several methods used for their analysis, most require long reaction times to form UV absorbing or fluorescent derivatives and the nonpolar nature of commonly used derivatives necessitates relatively high concentrations of organic solvents for elution in reverse phase liquid chromatography. The present method describes the use of Girard-T reagent in a simple, one step derivatization of α-dicarbonyls and conjugated aldehydes and analysis using ion-pair reverse phase liquid chromatography. The limit of detection was in the range of 0.06-0.09 μM (4-12 ng/mL) for glyoxal, methylglyoxal, 3-deoxyglucosone and 5-hydroxymethylfurfural with good linear response and reproducibility using UV detection. The hydrazone derivatives were stable for several days in solution. The method was used to study degradation of several sugars and quantification of the target α-dicarbonyls and 5-hydroxymethylfurfural in several soft drinks.
Topics: Betaine; Chromatography, Liquid; Deoxyglucose; Furaldehyde; Glyoxal; High Fructose Corn Syrup; Limit of Detection; Linear Models; Reproducibility of Results
PubMed: 28977384
DOI: 10.1093/chromsci/bmx073