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Am J Physiol Renal Physiol 277: F779-F789, 1999;
0363-6127/99 $5.00
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Vol. 277, Issue 5, F779-F789, November 1999

Inhibition of NFkappa B activation with antioxidants is correlated with reduced cytokine transcription in PTC

Gopala K. Rangan, Yiping Wang, Yuet-Ching Tay, and David C. H. Harris

Department of Renal Medicine, University of Sydney at Westmead Hospital, Westmead, Sydney, Australia 2145

We recently reported that inhibition of the transcription factor nuclear factor-kappa B (NFkappa B) with pyrrolidinedithiocarbamate (PDTC) reduced interstitial monocyte infiltration in rats with proteinuric tubulointerstitial disease, whereas N-acetylcysteine (NAC) was not effective. Here we investigate the effects of antioxidants (PDTC, NAC, and quercetin) on NFkappa B activation and cytokine transcription in primary cultured rat proximal tubular epithelial cells (PTC) stimulated with lipopolysaccharide. Antioxidant-mediated inhibition of NFkappa B activation (PDTC, 20-100 µM; NAC, 100 mM; and quercetin, 50 µM) diminished the induction of both pro- [interleukin (IL)-1beta , tumor necrosis factor-alpha , monocyte chemoattractant protein-1, macrophage inflammatory protein (MIP)-1alpha , and MIP-2] and anti-inflammatory (IL-10, transforming growth factor-beta 1) cytokine transcription in PTC (RT-PCR analysis). PDTC and quercetin did not affect PTC viability, but NAC (100 mM) caused a threefold increase in lactate dehydrogenase leakage (P < 0.001). We conclude that NAC is unable to suppress NFkappa B activation in PTC at subtoxic and physiologically relevant concentrations. Furthermore, antioxidant-mediated inhibition of NFkappa B is correlated with the nonselective reduction of cytokine transcription in activated tubular cells. These data might explain the protective effects of PDTC-mediated NFkappa B inhibition in tubulointerstitial disease in vivo.

N-acetylcysteine; pyrrolidinedithiocarbamate; quercetin; primary culture; osteopontin; proximal tubular cells; nuclear factor-kappa B


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