Apoptosis

, 16:1217 | Cite as

Glutamate dehydrogenase requirement for apoptosis induced by aristolochic acid in renal tubular epithelial cells

  • Victor Romanov
  • Terry Whyard
  • Radha Bonala
  • Francis Johnson
  • Arthur Grollman
Original Paper
First Online:

Abstract

Ingestion of aristolochic acids (AA) contained in herbal remedies results in a renal disease and, frequently, urothelial malignancy. The genotoxicity of AA in renal cells, including mutagenic DNA adduct formation, is well-documented. However, the mechanisms of AA-induced tubular atrophy and renal fibrosis are largely unknown. Epithelial cell death is a critical characteristic of these pathological conditions. To elucidate the mechanisms of AA-induced cytotoxicity, we explored AA-interacting proteins in tubular epithelial cells (TEC). We found that AA interacts with a mitochondrial enzyme glutamate dehydrogenase (GDH) and moderately inhibits its activity. We report that AA induces cell death in GDH-knockdown TEC preferentially via non-apoptotic means, whereas in GDH-positive cells, death was executed by both the non-apoptotic and apoptotic mechanisms. Apoptosis is an energy-reliant process and demands higher adenosine 5′-triphosphate (ATP) consumption than does the non-apoptotic cell death. We found that, after AAI treatment, the ATP depletion is more pronounced in GDH-knockdown cells. When we reduced ATP in TEC cells by inhibition of glycolysis and mitochondrial respiration, cell death mode switched from apoptosis and necrosis to necrosis only. In addition, in cells incubated at low glucose and no glutamine conditions, oxaloacetate and pyruvate reduced AAI-induced apoptosis our data suggest that AAI-GDH interactions in TEC are critical for the induction of apoptosis by direct inhibition of GDH activity. AA binding may also induce changes in GDH conformation and promote interactions with other molecules or impair signaling by GDH metabolic products, leading to apoptosis.

Keywords

Aristolochic acid Glutamate dehydrogenase Apoptosis Proximal tubules 

Abbreviations

NADP+

Nicotinamide adenine dinucleotide phosphate

ATP

Adenosine 5′-triphosphate

GTP

Guanosine 5′-Triphosphate

ADP

Adenosine diphosphate

SDS

Sodium dodecyl sulfate

PAGE

Polyacrylamide gel electrophoresis

shRNA

Short hairpin ribonucleic acid

OXPHOS

Oxidative phosphorylation

NAD

Nicotinamide adenine dinucleotide

EDTA

Ethylenediaminetetraacetic acid

DTT

Dithiothreitol

HEPES

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

EGCG

Epigallocatechin-3-monogallate

Gln

Glycine

DOG

2-deoxyglucose

DM-αKG

Dimethyl α ketoglutarate

FCCP

Cyanide-p-trifluoromethoxyphenylhydrazone

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Notes

Acknowledgments

We wish to thank Dr. Yury Lazebnik (Cold Spring Harbor Laboratory) for his valuable recommendations and insights concerning our data. We would also like to thank members of the Chemical Biology group (Department of Pharmacology, Stony Brook University) for sharing their perspective on the data. This study was supported by the NIH grant P01 ES004068 to A. G.

Conflict of interest

The authors declare no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Victor Romanov
    • 1
    • 2
  • Terry Whyard
    • 2
  • Radha Bonala
    • 1
  • Francis Johnson
    • 1
  • Arthur Grollman
    • 1
  1. 1.Department of Pharmacological SciencesState University of New York at Stony BrookStony BrookUSA
  2. 2.Department of UrologyState University of New York at Stony BrookStony BrookUSA

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