Glucose and pyruvate regulate cytokine-induced nitric oxide production by cardiac myocytes.
Metabolic requirements for the production of nitric oxide (NO) by cytokine-stimulated neonatal rat cardiac myocytes (CM) were studied. CM were cultured for 48 h in media containing interleukin-1 beta (IL-1 beta) and free fatty acids. Removal of glucose from the media partially inhibited IL-1 beta-stimulated nitrite (NO2-) production [8.1 +/- 0.3 vs. 4.4 +/- 0.6 nmol.(1.25 X 10(5) cells)-1.48 h-1; P < 0.01; n = 12]. The glycolytic inhibitor 2-deoxy-D-glucose (2-DG) completely inhibited IL-1 beta-stimulated NO2- production [0.7 +/- 0.5 nmol.(1.25 X 10(5) cells)-1.48 h-1; P < 0.01; n = 12]. The addition of the glycolytic end product, pyruvate, completely blocked the 2-DG inhibition of IL-1 beta-stimulated NO2- production [7.4 +/- 0.4 nmol.(1.25 X 10(5) cells)-1.48 h-1; P < 0.01; n = 12]. Pyruvate alone did not significantly enhance NO2- production in the presence or absence of glucose (n = 12). The inactive analogue 3-O-methylglucose had no effect on NO2- production (n = 12). Reverse transcription-polymerase chain reaction revealed that pyruvate blocked 2-DG inhibition of inducible NO synthase mRNA expression. Neither 2-DG nor pyruvate had any effect on GTP-cyclohydrolase I mRNA expression in CM. We report for the first time that optimal IL-1 beta-stimulated NO production by CM requires both glucose and the glycolytic end product pyruvate.
Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania 15213, USA.
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Finkel
D000818:Animals; D002478:Cells, Cultured; D003847:Deoxyglucose / Q000494:pharmacology; D006136:GTP Cyclohydrolase / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000502:physiology*; D006019:Glycolysis; D007375:Interleukin-1 / Q000494:pharmacology*; D009206:Myocardium / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000235:genetics; D011773:Pyruvates / Q000378:metabolism*; D051381:Rats
D007375:Interleukin-1; D011773:Pyruvates; D009569:Nitric Oxide; D003847:Deoxyglucose; D019001:Nitric Oxide Synthase; D006136:GTP Cyclohydrolase; D005947:Glucose