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Abstract
This work investigated the capacity of α-lipoic acid (LA) and N-acetyl-l-cysteine (NAC) to reduce zinc deficiency-induced oxidative stress, and prevent the activation of nuclear factor-κB (NF-κB) and activator protein-1 (AP-1), and the cross-talk between both activated cascades through β-Transducin Repeat-containing Protein (β-TrCP). IMR-32 cells were incubated in control media or media containing variable concentrations of zinc, without or with 0.5 mM LA or 1 mM NAC. Relative to control and zinc supplemented (15 μM Zn) groups, Hydrogen peroxide (H2O2) and total oxidant cell concentrations were higher, and total glutathione concentrations were lower in the zinc deficient groups (1.5 and 5 μM Zn). Both, LA and NAC, markedly reduced the increase in cell oxidants and the reduction in glutathione concentrations in the zinc deficient cells. Consistent with this, LA and NAC prevented zinc deficiency-induced activation of the early steps of NF- κB (IκBα phosphorylation) and AP-1 [c-Jun-N-terminal kinase (JNK) and p38 phophorylation] cascades, and the high NF-κB- and AP-1-DNA binding activities in total cell extracts. Thus, LA and NAC can reduce the oxidative stress associated with zinc deficiency and the subsequent triggering of NF-κB- and AP-1-activation in neuronal cells.
| Abbreviations | ||
| LA | = | ( ± )-α-lipoic acid |
| NAC | = | N-acetyl cysteine |
| AP-1 | = | activator protein-1 |
| NF-κB | = | nuclear factor-κB |
| JNK | = | c-Jun-N-terminal kinase |
| MAPKs | = | mitogenactivated protein kinases |
| DMEM | = | Dulbecco's modified Eagle medium |
| FBS | = | fetal bovine serum |
| EDTA | = | ethylenediamine tetraacetate |
| DTT | = | dithiothreitol |
| HEPES | = | 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide |
| β-TrCP | = | β-Transducin repeat-containing protein |
| DTPA | = | diethylenetriamine pentaacetic acid |
| PBS | = | phosphate-buffered saline |
| TSQ | = | N-6-(6-methoxy-8-quinolyl)-p-toluenesulfonamide |
| DCDHF | = | 5(or 6)-carboxy-2′7′-dichlorodihydrofluorescein diacetate |
| DCF | = | 5-(and-6)-carboxy-2′,7′-dichlorofluorescein |
| EMSA | = | electrophoretic mobility shift assay |
| Abbreviations | ||
| LA | = | ( ± )-α-lipoic acid |
| NAC | = | N-acetyl cysteine |
| AP-1 | = | activator protein-1 |
| NF-κB | = | nuclear factor-κB |
| JNK | = | c-Jun-N-terminal kinase |
| MAPKs | = | mitogenactivated protein kinases |
| DMEM | = | Dulbecco's modified Eagle medium |
| FBS | = | fetal bovine serum |
| EDTA | = | ethylenediamine tetraacetate |
| DTT | = | dithiothreitol |
| HEPES | = | 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid |
| MTT | = | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide |
| β-TrCP | = | β-Transducin repeat-containing protein |
| DTPA | = | diethylenetriamine pentaacetic acid |
| PBS | = | phosphate-buffered saline |
| TSQ | = | N-6-(6-methoxy-8-quinolyl)-p-toluenesulfonamide |
| DCDHF | = | 5(or 6)-carboxy-2′7′-dichlorodihydrofluorescein diacetate |
| DCF | = | 5-(and-6)-carboxy-2′,7′-dichlorofluorescein |
| EMSA | = | electrophoretic mobility shift assay |