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Abstract
This study was undertaken to determine if 4-hydroxy-2-nonenal (HNE) could upregulate antioxidants and phase 2 enzymes in rat H9c2 myocardiac cells, and if the upregulated defenses led to cytoprotection against oxidative and electrophilic injury. Incubation of H9c2 cells with HNE at noncytotoxic concentrations resulted in significant induction of cellular catalase, glutathione (GSH), GSH S-transferase (GST), and NAD(P)H:quinone oxidoreductase 1 (NQO1), as determined by enzyme activity and/or protein expression. HNE treatment caused increased mRNA expression of catalase, γ-glutamylcysteine ligase, GST-A1, and NQO1. Pretreatment of H9c2 cells with HNE led to significant protection against cytotoxicity induced by reactive oxygen and nitrogen species. HNE-pretreated cells also exhibited increased resistance to injury elicited by subsequent cytotoxic concentrations of HNE. Taken together, this study demonstrates that several antioxidants and phase 2 enzymes in H9c2 cells are upregulated by HNE and that the increased defenses afford protection against overt oxidative and electrophilic cardiac cell injury.
| Abbreviations | ||
| CDNB | = | 1-chloro-2,4-dinitrobenzene |
| DCFH | = | 2,7-dichlorodihydrofluorescein |
| DCIP | = | 2,6-dichloroindophenol |
| FBS | = | fetal bovine serum |
| GCL | = | γ-glutamylcysteine ligase |
| GPx | = | glutathione peroxidase |
| GR | = | glutathione reductase |
| GSH | = | reduced glutathione |
| GSSG | = | oxidized form of glutathione |
| GST | = | glutathione S-transferase |
| HNE | = | 4-hydroxy-2-nonenal |
| LDH | = | lactate dehydrogenase |
| MTT | = | 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide |
| NQO1 | = | NAD(P)H:quinone oxidoreductase-1 |
| PBS | = | phosphate buffered saline |
| RNS | = | reactive nitrogen species |
| ROS | = | reactive oxygen species |
| SIN-1 | = | 3-morpholinosydnonimine |
| SOD | = | superoxide dismutase |
| Abbreviations | ||
| CDNB | = | 1-chloro-2,4-dinitrobenzene |
| DCFH | = | 2,7-dichlorodihydrofluorescein |
| DCIP | = | 2,6-dichloroindophenol |
| FBS | = | fetal bovine serum |
| GCL | = | γ-glutamylcysteine ligase |
| GPx | = | glutathione peroxidase |
| GR | = | glutathione reductase |
| GSH | = | reduced glutathione |
| GSSG | = | oxidized form of glutathione |
| GST | = | glutathione S-transferase |
| HNE | = | 4-hydroxy-2-nonenal |
| LDH | = | lactate dehydrogenase |
| MTT | = | 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide |
| NQO1 | = | NAD(P)H:quinone oxidoreductase-1 |
| PBS | = | phosphate buffered saline |
| RNS | = | reactive nitrogen species |
| ROS | = | reactive oxygen species |
| SIN-1 | = | 3-morpholinosydnonimine |
| SOD | = | superoxide dismutase |