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Proinflammatory cytokines increase the rate of glycolysis and adenosine-5'-triphosphate turnover in cultured rat enterocytes
Linköping University, Faculty of Health Sciences. Linköping University, Department of Medicine and Health Sciences, Thoracic Surgery. Östergötlands Läns Landsting, Heart Centre, Department of Thoracic and Vascular Surgery.
Department of Critical Care Medicine, Univ. of Pittsburgh Sch. of Medicine, Pittsburgh, PA, United States.
Department of Critical Care Medicine, Univ. of Pittsburgh Sch. of Medicine, Pittsburgh, PA, United States.
Department of Critical Care Medicine, Univ. of Pittsburgh Sch. of Medicine, Pittsburgh, PA, United States, Department of Pathology, Univ. of Pittsburgh Sch. of Medicine, Pittsburgh, PA, United States.
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2003 (English)In: Critical Care Medicine, ISSN 0090-3493, Vol. 31, no 4, 1203-1212 p.Article in journal (Refereed) Published
Abstract [en]

Objective: Measurements of steady-state adenosine-5'-triphosphate (ATP) levels in tissue samples from patients or experimental animals with sepsis or endotoxemia provide little information about the rate of ATP production and consumption in these conditions. Accordingly, we sought to use an in vitro "reductionist" model of sepsis to test the hypothesis that proinflammatory cytokines modulate ATP turnover rate. Design: In vitro "reductionist" model of sepsis. Setting: University laboratory. Subjects: Cultured rat enterocyte-like cells. Interventions: IEC-6 nontransformed rat enterocytes were studied under control conditions or following incubation for 24 or 48 hrs with cytomix, a mixture of tumor necrosis factor-a (10 ng/mL), interleukin-1ß (1 ng/mL), and interferon-? (1000 units/mL). To measure ATP turnover rate, ATP synthesis was acutely blocked by adding to the cells a mixture of 2-deoxyglucose (10 mM), potassium cyanide (8 mM), and antimycin A (1 µM). ATP content was measured at baseline (before metabolic inhibition) and 0.5, 1, 2, 5, and 10 mins later. Log-linear ATP decay curves were generated and the kinetics of ATP utilization thereby calculated. Measurements and Main Results: ATP consumption rate was higher in cytomix-stimulated compared with control cells (3.11 ± 1.39 vs. 1.25 ± 0.66 nmol/min, respectively, p < .01). Similarly, the half-time for ATP disappearance was shorter in cytomix-stimulated compared with control cells (2.63 ± 1.00 vs. 6.21 ± 3.49, p < .05). In contrast to these findings, the rate of ATP disappearance was similar in cytokine-naïve and immunostimulated IEC-6 cells when protein and nucleic acid synthesis were inhibited by adding 50 µg/mL cycloheximide and 5 µg/mL actinomycin D to cultures for 4 hrs. The rates of glucose consumption and lactate production were significantly greater in cytomix-stimulated compared with controls cells. Conclusions: Incubation of IEC-6 cells with cytomix significantly increased ATP turnover. Increased ATP turnover rate was supported by increases in the rate of anaerobic glycolysis. These findings support the view that proinflammatory mediators impose a metabolic demand on visceral cells. In sepsis, cells may be more susceptible to dysfunction on the basis of diminished oxygen delivery and/or mitochondrial dysfunction.

Place, publisher, year, edition, pages
2003. Vol. 31, no 4, 1203-1212 p.
Keyword [en]
Energy metabolism, Glycolysis, Lactate, Nucleic acid synthesis, Protein synthesis
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-46686DOI: 10.1097/01.CCM.0000059647.92390.92OAI: oai:DiVA.org:liu-46686DiVA: diva2:267582
Available from: 2009-10-11 Created: 2009-10-11 Last updated: 2011-01-13

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Berg, Sören

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