Activation of NF-κB in Normal Rat Kidney Epithelial (NRK52E) Cells Is Mediated via a Redox-Insensitive, Calcium-Dependent Pathway

doi:10.1006/taap.1998.8583

Toxicology and Applied Pharmacology

Volume 154, Issue 3, 1 February 1999, Pages 219-227

https://doi.org/10.1006/taap.1998.8583 Get rights and content

Abstract

Renal tubular epithelial cells are largely resistant to oxidant-induced injury despite their capacity to accumulate relatively high concentrations of potentially damaging prooxidant and thiol-depleting agents. In the present study, we tested the hypothesis that such resistance may be attributable to a lack or deficiency of signaling transduction pathways through which reactive oxidants have been shown to promote the activation of NF-κB, a transcriptional factor that is known to mediate the inducible expression of a wide variety of genes that are involved in inflammatory and other cytotoxic reactions in numerous cell types. NF-κB was found to be readily activated following exposure of cultured normal rat kidney epithelial (NRK52E) cells to bacterial lipopolysaccharide (LPS). However, in contrast to findings with many other cell types, the activation of NF-κB by LPS was not substantially altered either by pretreatment of cells with the thiol antioxidant,N-acetylcysteine, or by glutathione (GSH) depletion. Moreover, reactive oxidants and oxidative stress-generating chemicals were completely without effect with respect to NF-κB activation in NRK52E cells, even following GSH depletion. In contrast, LPS activation of NF-κB was substantially attenuated by the intracellular Ca²⁺chelator, Quin 2AM, and by the Ca-channel inhibitor, ruthenium red. Moreover, thapsigargin, a Ca–ATPase inhibitor, promoted NF-κB activation comparable to that observed by LPS. Additionally, staurosporine, a Ca-dependent protein kinase C inhibitor, substantially decreased LPS-mediated NF-κB activation. These results demonstrate that the LPS-inducible expression of NF-κB in renal epithelial cells, in contrast to many other cell types, is not responsive to oxidative stress and is regulated, at least in part, by redox-insensitive modulation of intracellular calcium levels. These findings provide a basis for the highly tissue-specific expression and function of NF-κB in kidney epithelial cells, which may underlie their resistance to oxidant-mediated cytotoxicity.

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Dithiocarbamates can modulate the expression of genes associated with inflammation or development of ischemia/reperfusion injury. Here, we investigate the effects of pyrrolidine dithiocarbamate, an inhibitor of nuclear factor (NF)-κB activation, on the renal dysfunction and injury caused by ischemia/reperfusion of the rat kidney. Bilateral clamping of renal pedicles (45 min) followed by reperfusion (6 h) caused significant renal dysfunction and marked renal injury. Pyrrolidine dithiocarbamate (100 mg/kg, administered i.v.) significantly reduced biochemical and histological evidence of renal dysfunction and injury caused by ischemia/reperfusion of the rat kidney. Furthermore, pyrrolidine dithiocarbamate markedly reduced the expression of inducible nitric oxide synthase (iNOS) protein and significantly reduced serum levels of nitric oxide. Finally, pyrrolidine dithiocarbamate inhibited the activation of NF-κB by preventing its translocation from the cytoplasm into the nuclei of renal cells. These results demonstrate that pyrrolidine dithiocarbamate reduces renal ischemia/reperfusion injury and that dithiocarbamates may provide beneficial actions against ischemic acute renal failure.
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Regular ArticleActivation of NF-κB in Normal Rat Kidney Epithelial (NRK52E) Cells Is Mediated via a Redox-Insensitive, Calcium-Dependent Pathway

Abstract

Kidney Int.

Cell

Biochim. Biophys. Acta

J. Biol. Chem.

Free Radical Biol. Med.

Cell

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

Cancer Biol.

J. Biol. Chem.

Immunobiology

Free Radical Biol. Med.

Biochim. Biophys. Acta

Blood

FEBS Lett.

Cell

Arch. Biochem. Biophys.

J. Biol. Chem.

IκB: a specific inhibitor of the NF-κB transcription factor

Science (Washington, D.C.)

Function and activation of NF-κB in the immune system

Annu. Rev. Immunol.

Flow cytometric studies of oxidative product formation by neutrophils: A graded response to membrane stimulation

J. Immunol.

Inhibition of NF-κB activity induces apoptosis in murine hepatocytes

Am. J. Pathol.

A role for transcription factor NF-κB in inflammation

Inflam. Res.

Differential mechanisms of LPS-induced NF-κB activation in macrophages and fibroblasts

J. Endotox. Res.

Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei

Nucleic Acid Res.

Constitutive nuclear NF-κB in cells of monocyte lineage

Biochem. J.

Regular Article
Activation of NF-κB in Normal Rat Kidney Epithelial (NRK52E) Cells Is Mediated via a Redox-Insensitive, Calcium-Dependent Pathway