Ginger Compound [6]-Shogaol and Its Cysteine-Conjugated Metabolite (M2) Activate Nrf2 in Colon Epithelial Cells in Vitro and in Vivo
| Autor: | Edward E. Schmidt, Feng Yan, Xiaoxin Chen, Michael B. Major, Junsheng Fu, Justin R. Prigge, Shengmin Sang, Dominique N. Soroka, Huadong Chen, Hao Chen, Yuhui Hu |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Alkylation
NF-E2-Related Factor 2 Glutamate-Cysteine Ligase Catechols Biology Ginger Toxicology environment and public health Article 03 medical and health sciences chemistry.chemical_compound Mice 0302 clinical medicine Downregulation and upregulation In vivo Animals Humans Cysteine Phosphorylation Protein Kinase Inhibitors 030304 developmental biology Mice Knockout 0303 health sciences Kelch-Like ECH-Associated Protein 1 GCLM Intracellular Signaling Peptides and Proteins Membrane Proteins Epithelial Cells General Medicine Shogaol Glutathione respiratory system HCT116 Cells Molecular biology In vitro Up-Regulation GCLC chemistry Biochemistry 030220 oncology & carcinogenesis Reactive Oxygen Species Intracellular Heme Oxygenase-1 |
| Zdroj: | Chemical Research in Toxicology |
| ISSN: | 1520-5010 0893-228X |
| Popis: | In this study, we identified Nrf2 as a molecular target of [6]-shogaol (6S), a bioactive compound isolated from ginger, in colon epithelial cells in vitro and in vivo. Following 6S treatment of HCT-116 cells, the intracellular GSH/GSSG ratio was initially diminished but was then elevated above the basal level. Intracellular reactive oxygen species (ROS) correlated inversely with the GSH/GSSG ratio. Further analysis using gene microarray showed that 6S upregulated the expression of Nrf2 target genes (AKR1B10, FTL, GGTLA4, and HMOX1) in HCT-116 cells. Western blotting confirmed upregulation, phosphorylation, and nuclear translocation of Nrf2 protein followed by Keap1 decrease and upregulation of Nrf2 target genes (AKR1B10, FTL, GGTLA4, HMOX1, and MT1) and glutathione synthesis genes (GCLC and GCLM). Pretreatment of cells with a specific inhibitor of p38 (SB202190), PI3K (LY294002), or MEK1 (PD098059) attenuated these effects of 6S. Using ultra-high-performance liquid chromatography-tandem mass spectrometry, we found that 6S modified multiple cysteine residues of Keap1 protein. In vivo 6S treatment induced Nrf2 nuclear translocation and significantly upregulated the expression of MT1, HMOX1, and GCLC in the colon of wild-type mice but not Nrf2(-/-) mice. Similar to 6S, a cysteine-conjugated metabolite of 6S (M2), which was previously found to be a carrier of 6S in vitro and in vivo, also activated Nrf2. Our data demonstrated that 6S and its cysteine-conjugated metabolite M2 activate Nrf2 in colon epithelial cells in vitro and in vivo through Keap1-dependent and -independent mechanisms. |
| Databáze: | OpenAIRE |
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