Our understanding of the interleukin-1 (IL-1) signaling molecular mechanisms has recently made considerable progress, with the discovery of the IL-1 receptor-associated kinase and the downstream enzymatic cascade that leads to the activation of nuclear factor-κB (NF-κB). IL-1 signaling and especially NF-κB activation are thought to be redox-sensitive, even though the precise nature and the molecular targets of the oxidants/antioxidants involved remain largely unknown. Here, we investigated the possible role of cellular oxidized/reduced glutathione (GSSG/GSH) balance in IL-1 signaling. We describe a quantitative method based on capillary electrophoresis designed to assay both intracellular GSH and GSSG in adhering fibroblasts. This method allows the GSSG/GSH balance to be followed during IL-1 stimulation. Our data show that IL-1 induces rapid and transient oxidation of intracellular glutathione in human fibroblasts. Using various antioxidants, including pyrrolidine dithiocarbamate and curcumin, we were unable to show a direct relationship between this IL-1-induced glutathione oxidation and NF-κB activation. Of the five antioxidants tested, only curcumin was able to inhibit IκBα degradation upstream and, hence, NF-κB DNA-binding activity and NF-κB-dependent expression of IL-6 downstream.
|Number of pages||12|
|Journal||Antioxidants and Redox Signaling|
|Publication status||Published - Apr 2001|