scholarly journals Response of C2C12 Myoblasts to Hypoxia: The Relative Roles of Glucose and Oxygen in Adaptive Cellular Metabolism

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Wei Li ◽  
Zhen-Fu Hu ◽  
Bin Chen ◽  
Guo-Xin Ni
Keyword(s):  
Oxygen Consumption ◽  
Glucose Concentration ◽  
Cellular Response ◽  
Cell Function ◽  
Cell Metabolism ◽  
Lactate Production ◽  
Cellular Metabolism ◽  
Glucose Consumption ◽  
C2c12 Cells ◽  
Low Oxygen

Background. Oxygen and glucose are two important nutrients for mammalian cell function. In this study, the effect of glucose and oxygen concentrations on C2C12 cellular metabolism was characterized with an emphasis on detecting whether cells show oxygen conformance (OC) in response to hypoxia.Methods. After C2C12 cells being cultured in the levels of glucose at 0.6 mM (LG), 5.6 mM (MG), or 23.3 mM(HG) under normoxic or hypoxic (1% oxygen) condition, cellular oxygen consumption, glucose consumption, lactate production, and metabolic status were determined. Short-term oxygen consumption was measured with a novel oxygen biosensor technique. Longer-term measurements were performed with standard glucose, lactate, and cell metabolism assays.Results. It was found that oxygen depletion in normoxia is dependent on the glucose concentration in the medium. Cellular glucose uptake and lactate production increased significantly in hypoxia than those in normoxia. In hypoxia the cellular response to the level of glucose was different to that in normoxia. The metabolic activities decreased while glucose concentration increased in normoxia, while in hypoxia, metabolic activity was reduced in LG and MG, but unchanged in HG condition. The OC phenomenon was not observed in the present study.Conclusions. Our findings suggested that a combination of low oxygen and low glucose damages the viability of C2C12 cells more seriously than low oxygen alone. In addition, when there is sufficient glucose, C2C12 cells will respond to hypoxia by upregulating anaerobic respiration, as shown by lactate production.

scholarly journals Metabolic Signatures of Cryptosporidium parvum-Infected HCT-8 Cells and Impact of Selected Metabolic Inhibitors on C. parvum Infection under Physioxia and Hyperoxia

Biology ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 60
Author(s):  
Juan Vélez ◽  
Zahady Velasquez ◽  
Liliana M. R. Silva ◽  
Ulrich Gärtner ◽  
Klaus Failing ◽  
...  
Keyword(s):  
Host Cell ◽  
Cryptosporidium Parvum ◽  
Cell Metabolism ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Host Cells ◽  
Metabolic Inhibitors ◽  
Low Oxygen ◽  
Metabolic Signatures

Cryptosporidium parvum is an apicomplexan zoonotic parasite recognized as the second leading-cause of diarrhoea-induced mortality in children. In contrast to other apicomplexans, C.parvum has minimalistic metabolic capacities which are almost exclusively based on glycolysis. Consequently, C. parvum is highly dependent on its host cell metabolism. In vivo (within the intestine) infected epithelial host cells are typically exposed to low oxygen pressure (1–11% O2, termed physioxia). Here, we comparatively analyzed the metabolic signatures of C. parvum-infected HCT-8 cells cultured under both, hyperoxia (21% O2), representing the standard oxygen condition used in most experimental settings, and physioxia (5% O2), to be closer to the in vivo situation. The most pronounced effect of C. parvum infection on host cell metabolism was, on one side, an increase in glucose and glutamine uptake, and on the other side, an increase in lactate release. When cultured in a glutamine-deficient medium, C. parvum infection led to a massive increase in glucose consumption and lactate production. Together, these results point to the important role of both glycolysis and glutaminolysis during C. parvum intracellular replication. Referring to obtained metabolic signatures, we targeted glycolysis as well as glutaminolysis in C. parvum-infected host cells by using the inhibitors lonidamine [inhibitor of hexokinase, mitochondrial carrier protein (MCP) and monocarboxylate transporters (MCT) 1, 2, 4], galloflavin (lactate dehydrogenase inhibitor), syrosingopine (MCT1- and MCT4 inhibitor) and compound 968 (glutaminase inhibitor) under hyperoxic and physioxic conditions. In line with metabolic signatures, all inhibitors significantly reduced parasite replication under both oxygen conditions, thereby proving both energy-related metabolic pathways, glycolysis and glutaminolysis, but also lactate export mechanisms via MCTs as pivotal for C. parvum under in vivo physioxic conditions of mammals.

scholarly journals Distinct profiles of human embryonic stem cell metabolism and mitochondria identified by oxygen

Reproduction ◽  
2015 ◽  
Vol 150 (4) ◽  
pp. 367-382 ◽  
Author(s):  
Jarmon G Lees ◽  
Joy Rathjen ◽  
John R Sheedy ◽  
David K Gardner ◽  
Alexandra J Harvey
Keyword(s):  
Amino Acid ◽  
Mitochondrial Function ◽  
Cell Function ◽  
Atmospheric Oxygen ◽  
Cell Metabolism ◽  
Embryonic Stem ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Mitochondrial Activity ◽  
Hes Cells

Oxygen is a powerful regulator of cell function and embryonic development. It has previously been determined that oxygen regulates human embryonic stem (hES) cell glycolytic and amino acid metabolism, but the effects on mitochondria are as yet unknown. Two hES cell lines (MEL1, MEL2) were analyzed to determine the role of 5% (physiological) and 20% (atmospheric) oxygen in regulating mitochondrial activity. In response to extended physiological oxygen culture, MEL2 hES cells displayed reduced mtDNA content, mitochondrial mass and expression of metabolic genesTFAM,NRF1,PPARaandMT-ND4. Furthermore, MEL2 hES cell glucose consumption, lactate production and amino acid turnover were elevated under physiological oxygen. In stark contrast, MEL1 hES cell amino acid and carbohydrate use and mitochondrial function were relatively unaltered in response to oxygen. Furthermore, differentiation kinetics were delayed in the MEL1 hES cell line following BMP4 treatment. Here we report the first incidence of metabolic dysfunction in a hES cell population, defined as a failure to respond to oxygen concentration through the modulation of metabolism, demonstrating that hES cells can be perturbed during culture despite exhibiting the defining characteristics of pluripotent cells. Collectively, these data reveal a central role for oxygen in the regulation of hES cell metabolism and mitochondrial function, whereby physiological oxygen promotes glucose flux and suppresses mitochondrial biogenesis and gene expression.

ÉTUDE IN VITRO DU MÉTABOLISME DES HYDRATES DE CARBONE DANS LE LEUCOCYTE CIRCULANT DU COBAYE TRAITÉ À LA CORTISONE

1966 ◽  
Vol 51 (2) ◽  
pp. 193-202
Author(s):  
J. A. Antonioli ◽  
A. Vannotti
Keyword(s):  
Glucose Concentration ◽  
Intramuscular Injection ◽  
Acute Inflammation ◽  
Glycogen Content ◽  
Glycogen Synthesis ◽  
Lactate Production ◽  
Glucose Consumption ◽  
List Type ◽  
Hydrates De Carbone

ABSTRACT 1. The metabolism of suspensions of circulating leucocytes has been studied after intramuscular injection of a dose of 50 mg/kg of a corticosteroid (cortisone acetate). The suspensions were incubated under aerobic conditions in the presence of a glucose concentration of 5.6 mm. Glucose consumption, lactate production, and variations in intracellular glycogen concentration were measured. After the administration of the corticosteroid, the anabolic processes of granulocyte metabolism were reversibly stimulated. Glucose consumption and lactate production increased 12 hours after the injection, but tended to normalize after 24 hours. The glycogen content of the granulocytes was enhanced, and glycogen synthesis during the course of the incubation was greatly stimulated. The action of the administered corticosteroid is more prolonged in females than in males. The injection of the corticosteroid caused metabolic modifications which resemble in their modulations and in their chronological development those found in circulating granulocytes of guinea-pigs suffering from sterile peritonitis. These results suggest, therefore, that, in the case of acute inflammation, the glucocorticosteroids may play an important role in the regulation of the metabolism of the blood leucocytes.

LEUCOCYTE METABOLISM IN PREGNANCY

1960 ◽  
Vol XXXV (IV) ◽  
pp. 575-584 ◽  
Author(s):  
C. Borel ◽  
J. Frei ◽  
A. Vannotti
Keyword(s):  
Alkaline Phosphatase ◽  
Oxygen Consumption ◽  
Pregnant Women ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Lactate Production ◽  
Glucose Consumption ◽  
In Pregnancy

ABSTRACT Enzymatic studies, on leucocytes of pregnant women, show an increase of the alkaline phosphatase activity and a decrease of the glucose consumption and lactate production, as well as of proteolysis. The oxygen consumption, with succinate as substrate, does not vary.

scholarly journals Metabolic regulation of collagen gel contraction by porcine aortic valvular interstitial cells

2014 ◽  
Vol 11 (101) ◽  
pp. 20140852 ◽  
Author(s):  
Peter I. Kamel ◽  
Xin Qu ◽  
Andrew M. Geiszler ◽  
Deepak Nagrath ◽  
Romain Harmancey ◽  
...  
Keyword(s):  
Metabolic Regulation ◽  
Heart Valves ◽  
Cell Metabolism ◽  
Lactate Production ◽  
Collagen Gel ◽  
Glucose Consumption ◽  
Mechanical Stimuli ◽  
Collagen Gels ◽  
Calcific Aortic Valve Disease ◽  
Metabolic Substrates

Despite a high incidence of calcific aortic valve disease in metabolic syndrome, there is little information about the fundamental metabolism of heart valves. Cell metabolism is a first responder to chemical and mechanical stimuli, but it is unknown how such signals employed in valve tissue engineering impact valvular interstitial cell (VIC) biology and valvular disease pathogenesis. In this study porcine aortic VICs were seeded into three-dimensional collagen gels and analysed for gel contraction, lactate production and glucose consumption in response to manipulation of metabolic substrates, including glucose, galactose, pyruvate and glutamine. Cell viability was also assessed in two-dimensional culture. We found that gel contraction was sensitive to metabolic manipulation, particularly in nutrient-depleted medium. Contraction was optimal at an intermediate glucose concentration (2 g l −1 ) with less contraction with excess (4.5 g l −1 ) or reduced glucose (1 g l −1 ). Substitution with galactose delayed contraction and decreased lactate production. In low sugar concentrations, pyruvate depletion reduced contraction. Glutamine depletion reduced cell metabolism and viability. Our results suggest that nutrient depletion and manipulation of metabolic substrates impacts the viability, metabolism and contractile behaviour of VICs. Particularly, hyperglycaemic conditions can reduce VIC interaction with and remodelling of the extracellular matrix. These results begin to link VIC metabolism and macroscopic behaviour such as cell–matrix interaction.

Effect of glucose supply on ovine uteroplacental glucose metabolism

1999 ◽  
Vol 277 (4) ◽  
pp. R947-R958 ◽  
Author(s):  
Peter W. Aldoretta ◽  
William W. Hay
Keyword(s):  
Oxygen Consumption ◽  
Glucose Metabolism ◽  
Glucose Oxidation ◽  
Lactate Production ◽  
Glucose Consumption ◽  
Reciprocal Relationship ◽  
Late Gestation ◽  
Fick Principle ◽  
Pregnant Sheep ◽  
Glucose Supply

To test the hypothesis that glucose supply to the uteroplacenta (UP) regulates UP glucose metabolism into oxidative and nonoxidative pathways, we studied eight late-gestation pregnant sheep at low (LG) and high (HG) maternal glucose concentrations (GM), using Fick principle and tracer glucose methodology. UP glucose consumption (UPGC) correlated directly with GM( r = 0.75, P = 0.0006), and UP glucose decarboxylation ( r = 0.80, P = 0.0001), and lactate production ( r = 0.90, P = 0.0001) rates were directly correlated with UPGC. The combined fractional production rate for lactate, fructose, and CO2 from UPGC was the same in LG and HG periods. The fraction of UP oxygen consumption used for glucose oxidation increased by about 50% from LG to HG conditions; however, there was no change in UP oxygen consumption. Nearly half of UPGC was not accounted for by lactate, fructose, and CO2 production, and about two-thirds of UP oxygen consumption was not accounted for by immediate oxidation of glucose carbon just taken up by the UP. These results indicate that glucose supply directly regulates UP glucose oxidative metabolism and that there is a reciprocal relationship between UP glucose oxidation and the oxidation of other substrates.

EFFECTS OF EXPERIMENTAL HYPOTHYROIDISM ON SKELETAL MUSCLE METABOLISM IN THE RAT

1978 ◽  
Vol 87 (1) ◽  
pp. 114-124 ◽  
Author(s):  
C. van Hardeveld ◽  
A. A. H. Kassenaar
Keyword(s):  
Skeletal Muscle ◽  
Oxygen Consumption ◽  
Initial Level ◽  
Control Level ◽  
Lactate Production ◽  
Muscle Metabolism ◽  
Glucose Consumption ◽  
Glycerol Production ◽  
Base Level ◽  
Limb Perfusion

ABSTRACT Hind-limb perfusion was used to study the effect of thyroidectomy on some metabolic parameters in the skeletal muscle of the rat. A week after thyroidectomy obtained by one dose of 3/4 mCi 131I, neither T4 nor T3 was detected in the blood. Lactate production and glycerol production were already decreased a week after the treatment and reached a base level at two weeks. At that time, the oxygen consumption was significantly lower (70 % of initial level) than in the control animals and decreased further in the third week to nearly 50 % of the control level. Glucose consumption and alanine release were decreased three weeks after thyroidectomy. One dose of T3 (10 μg/100 g b. w.), administered to animals two weeks after the injection of 131I, restored the oxygen consumption, lactate production, and glycerol production to normal levels in 24 h. After 48 h, the glucose consumption was normal. Glycerol production was already significantly increased 6 h after T3 injection in animals one week after thyroidectomy, and in another group of animals two weeks after thyroidectomy. Apparently the diminished oxygen consumption in the latter group does not retard the lipolytic response to T3. No direct relationship could be found between the activity of lipolytic process and the thyroid hormone controlled oxygen consumption.

scholarly journals Effects of Dental Methacrylates on Oxygen Consumption and Redox Status of Human Pulp Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Giuseppina Nocca ◽  
Cinzia Callà ◽  
Giuseppe Ettore Martorana ◽  
Loredana Cicillini ◽  
Sandro Rengo ◽  
...  
Keyword(s):  
Oxygen Consumption ◽  
Consumption Rate ◽  
Lactate Production ◽  
Triethylene Glycol ◽  
Glucose Consumption ◽  
Redox Status ◽  
Triethylene Glycol Dimethacrylate ◽  
Pulp Cells ◽  
Species Production ◽  
Glucose 6 Phosphate Dehydrogenase

Several studies have already demonstrated that the incomplete polymerization of resin-based dental materials causes the release of monomers which might affect cell metabolism. The aim of this study was to investigate the effects of triethylene glycol dimethacrylate, 1,4-butanediol dimethacrylate, urethane dimethacrylate, and 2-hydroxyethyl methacrylate on (1) cellular energy metabolism, evaluating oxygen consumption rate, glucose consumption, glucose 6-phosphate dehydrogenase activity, and lactate production, and (2) cellular redox status, through the evaluation of glutathione concentration and of the activities of enzymes regulating glutathione metabolism.Methods. Human pulp cells were used and oxygen consumption was measured by means of a Clark electrode. Moreover, reactive oxygen species production was quantified. Enzymatic activity and glucose and lactate concentrations were determined through a specific kit.Results. Triethylene glycol dimethacrylate, 1,4-butanediol dimethacrylate, and 2-hydroxyethyl methacrylate induced a decrease in oxygen consumption rate, an enhancement of glucose consumption, and lactate production, whilst glucose 6-phosphate dehydrogenase and glutathione reductase activity were not significantly modified. Moreover, the monomers induced an increase of reactive oxygen species production with a consequent increase of superoxide dismutase and catalase enzymatic activities. A depletion of both reduced and total glutathione was also observed.Conclusion. The obtained results indicate that dental monomers might alter energy metabolism and glutathione redox balance in human pulp cells.

INFLUENCE OF EXPERIMENTAL HYPERTHYROIDISM ON SKELETAL MUSCLE METABOLISM IN THE RAT

1977 ◽  
Vol 85 (1) ◽  
pp. 71-83 ◽  
Author(s):  
C. van Hardeveld ◽  
A. A. H. Kassenaar
Keyword(s):  
Skeletal Muscle ◽  
Oxygen Consumption ◽  
Dose Range ◽  
Nervous Activity ◽  
Lactate Production ◽  
Muscle Metabolism ◽  
Glucose Consumption ◽  
Daily Injection ◽  
Limb Perfusion ◽  
Energy Consuming

ABSTRACT In this study hind-limb perfusion was used to investigate the influence of thyroid hormones on some metabolic parameters in the skeletal muscle of the rat. Daily injection of 20 μg L-thyroxine (T4) per 100 g b. w. for a week caused a 25 % increase in oxygen consumption. Further enlargement of the T4 dose had little additive effect. In the dose range 20–80 μg T4/100 g b. w., no important changes occurred in lactate production or glucose consumption. Only at the highest T4 dose did the glucose consumption increase significantly. The most profound effect of T4 was on lipolysis. A daily dose of 20 μg T4/100 g b. w. gave a doubling of glycerol production rate, the maximum occurring at a dose of 40 μg T4/100 g b. w Inactivation of the nervous system was without influence on the T4-induced increase in oxygen consumption. However, the T4-induced elevation of lipolysis disappeared after abolition of the nervous activity. This raises the possibility that the T4 effect on lipolysis in skeletal muscle is a potentiation of catecholamine effects. The T4-induced oxygen consumption increase might be dependent not on the lipolytic process but rather on other energy-consuming cell processes.

Online Measurement of Glucose Consumption from HepG2 Cells Using an Integrated Bioreactor and Enzymatic Assay

2018 ◽  
Author(s):  
Anna Adams ◽  
Radha Krishna Murthy Bulusu ◽  
Nikita Mukhitov ◽  
Jose Mendoza-Cortes ◽  
Michael Roper
Keyword(s):  
Real Time ◽  
Glucose Concentration ◽  
Hepg2 Cells ◽  
Glucose Consumption ◽  
Microfluidic System ◽  
Structure And Function ◽  
Online Measurement ◽  
Fluorescent Assay ◽  
And Function ◽  
Integrated Bioreactor

In this work, we developed a microfluidic bioreactor for optimizing growth and maintaining structure and function of HepG2, and when desired, the device could be removed and the extracellular output from the bioreactor combined with enzymatic glucose reagents into a droplet-based microfluidic system. The intensity of the resulting fluorescent assay product in the droplets was measured, and was directly correlated to glucose concentration, allowing the effect of insulin on glucose consumption in the HepG2 cells to be observed and quantified online and in near real-time.

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