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Thiol modulation of TNFα and IL-1 induced MnSOD gene expression and activation of NF-κB

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Abstract

TNFα and IL-1 each can activate NF-κB and induce gene expression of manganese superoxide dismutase (MnSOD), a mitochondrial matrix enzyme which can provide critical protection against hyperoxic lung injury. The regulation of MnSOD gene expression is not well understood. Since redox status can modulate NF-κB and potential κB site(s) exist in the MnSOD promoter, the effect of thiols (including NAC, DTT and 2-ME) on TNFα and IL-1 induced activation of NF-κB and MnSOD gene expression was investigated. Activation of NF-kB and increased MnSOD expression were potentiated by thiol reducing agents. In contrast, thiol oxidizing or alkylating agents inhibited both NF-κB activation and elevated MnSOD expression in response to TNFα or IL-1. Since protease inhibitors TPCK and TLCK can inhibit NF-κB activation, we also investigated the effect of these compounds on MnSOD expression and NF-κB activation. TPCK and TLCK each inhibited MnSOD gene expression and NF-κB activation. Since the MnSOD promoter also contains anAP-1 binding site, the effect of thiols and thiol modifying agents on AP-1 activation was investigated. Thiols had no consistent effect onAP-1 activation. Likewise, some of the thiol modifying compounds inhibited AP-1 activation by TNFα or IL-1, whereas others did not. Since diverse agents had similar effects on activation of NF-κB and MnSOD gene expression, we have demonstrated that activation of NF-κB and MnSOD gene expression are closely associated and that reduced sulfhydryl groups are required for cytokine mediation of both processes.

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Abbreviations

O2 :

Superoxide radical

H2O2 :

Hydrogen peroxide

NAC:

N-acetyl L-cysteine

DTT:

Dithiothreitol

2-ME:

2-Mercaptoethanol

MnSOD:

Manganese superoxide dismutase

NF-κB:

Nuclear factor kappa B

AP-1:

Activator protein-1

NBT:

Nitroblue tetrazolium

CAT:

Chloramphenicol acetyltransferase

TPCK:

N-tosyl-L-phenylalanine chloromethyl ketone

TLCK:

Na-p-tosyl-L-lysine chloromethyl ketone

TAME:

N-α-p-tosyl-L-arginine methyl ester

NEM:

N-ethyl maleimide

DEM:

Diethyl maleate

CDNB:

1-chloro-2,4-dinitrobenzene

DTTOX :

Oxidized dithiothreitol

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  1. Department of Pediatrics, National Jewish Center for Immunology and Respiratory Medicine, 80206, Denver, Colorado, USA

    Kumuda C. Das, Yvette Lewis-Molock & Carl W. White

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  2. Yvette Lewis-Molock
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Das, K.C., Lewis-Molock, Y. & White, C.W. Thiol modulation of TNFα and IL-1 induced MnSOD gene expression and activation of NF-κB. Mol Cell Biochem 148, 45–57 (1995). https://doi.org/10.1007/BF00929502

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