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Abstract
This study was undertaken to investigate the inducibility of glutathione (GSH), glutathione reductase (GR) and glutathione peroxidase (GPx) by 3H-1,2-dithiole-3-thione (D3T) in beta-cells, and the resultant cytoprotection against oxidant injury. Incubation of the insulin-secreting RINm5F cells with D3T led to significant induction of GSH, GR and GPx. D3T-mediated induction of GSH was abolished by buthionine sulfoximine (BSO), suggesting a critical involvement of γ-glutamylcysteine ligase (γGCL). Consistently, incubation of RINm5F cells with D3T resulted in increased expression of γGCL protein and mRNA. Pretreatment of RINm5F cells with D3T provided remarkable protection against oxidant-elicited cytotoxicity. On the other hand, depletion of cellular GSH by BSO sensitized RINm5F cells to oxidant injury. Furthermore, cotreatment of RINm5F cells with BSO to reverse D3T-mediated GSH induction abolished the cytoprotective effects of D3T on oxidant injury. Taken together, this study demonstrates that upregulation of glutathione system by D3T is effective for protecting against oxidative beta-cell injury.
| Abbreviations | ||
| BSO | = | buthionine sulfoximine |
| D3T | = | 3H-1,2-dithiole-3-thione |
| FBS | = | fetal bovine serum |
| γGCL | = | γ-glutamylcysteine ligase |
| γGCL-C | = | γ-glutamylcysteine ligase catalytic subunit |
| GPx | = | glutathione peroxidase |
| GR | = | glutathione reductase |
| GSH | = | rduced glutathione |
| GSSG | = | oxidized form of glutathione |
| LDH | = | lactate dehydrogenase |
| MTT | = | 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium |
| PBS | = | phosphate buffered saline |
| ROS | = | reactive oxygen species |
| RNS | = | reactive nitrogen species |
| SIN-1 | = | 3-morpholinosydnonimine |
| Abbreviations | ||
| BSO | = | buthionine sulfoximine |
| D3T | = | 3H-1,2-dithiole-3-thione |
| FBS | = | fetal bovine serum |
| γGCL | = | γ-glutamylcysteine ligase |
| γGCL-C | = | γ-glutamylcysteine ligase catalytic subunit |
| GPx | = | glutathione peroxidase |
| GR | = | glutathione reductase |
| GSH | = | rduced glutathione |
| GSSG | = | oxidized form of glutathione |
| LDH | = | lactate dehydrogenase |
| MTT | = | 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium |
| PBS | = | phosphate buffered saline |
| ROS | = | reactive oxygen species |
| RNS | = | reactive nitrogen species |
| SIN-1 | = | 3-morpholinosydnonimine |