Abstract
As part of a study on cadmium nephrotoxicity, we studied the effect of cadmium chloride (CdCl2) in isolated human renal proximal tubules metabolizing the physiological substrate lactate. Dose–effect experiments showed that 10–500 μM CdCl2 reduced lactate removal, glucose production and the cellular levels of ATP, coenzyme A, acetyl-coenzyme A and of reduced glutathione in a dose-dependent manner. After incubation with 5 mM l-[1-13C]-, or l-[2-13C]-, or l-[3-13C] lactate or 5 mM l-lactate plus 25 mM NaH13CO3 as substrates, substrate utilization and product formation were measured by both enzymatic and carbon 13 NMR methods. Combination of enzymatic and NMR measurements with a mathematical model of lactate metabolism previously validated showed that 100 μM CdCl2 caused an inhibition of flux through lactate dehydrogenase and alanine aminotransferase and through the entire gluconeogenic pathway; fluxes were diminished by 19% (lactate dehydrogenase), 28% (alanine aminotransferase), 28% (pyruvate carboxylase), 42% (phosphoenolpyruvate carboxykinase), and 52% (glucose-6-phosphatase). Such effects occurred without altering the oxidation of the lactate carbons or fluxes through enzymes of the tricarboxylic acid cycle despite a large fall of the cellular ATP level, a marker of the energy status and of the viability of the renal cells. These results that were observed at clinically relevant tissue concentrations of cadmium provide a biochemical basis for a better understanding of the cellular mechanism of cadmium-induced renal proximal tubulopathy in humans chronically exposed to cadmium.
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Acknowledgments
This work was financially supported by the CEA, INSERM, INRA and CNRS by the “ToxNuc-E” program. The authors thank J.L. Imbert and D. Gonnet (service central d’analyse du CNRS à Solaize) for the determination of cadmium content in the proximal tubules.
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Faiz, H., Conjard-Duplany, A., Boghossian, M. et al. Cadmium chloride inhibits lactate gluconeogenesis in isolated human renal proximal tubules: a cellular metabolomic approach with 13C-NMR. Arch Toxicol 85, 1067–1077 (2011). https://doi.org/10.1007/s00204-010-0633-6
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DOI: https://doi.org/10.1007/s00204-010-0633-6