Modulation of Nrf2/Keap1 system by Wasabi 6-methylthiohexyl isothiocyanate in ARE-mediatedNQO1expression
| Autor: | Masashi Nagai, Shunsuke Tanigawa, Si Qin, Yosuke Daikoku, Yoshimi Korenori, Takuma Kumamoto, Koji Miyamori, De-Xing Hou |
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| Rok vydání: | 2013 |
| Předmět: |
Wasabia
Small interfering RNA NF-E2-Related Factor 2 Electrophoretic Mobility Shift Assay Response Elements digestive system environment and public health Antioxidants chemistry.chemical_compound Transactivation Genes Reporter Isothiocyanates NAD(P)H Dehydrogenase (Quinone) Humans Electrophoretic mobility shift assay Regulation of gene expression Gene knockdown Kelch-Like ECH-Associated Protein 1 Intracellular Signaling Peptides and Proteins Hep G2 Cells respiratory system Molecular biology KEAP1 Gene Expression Regulation chemistry Cytoprotection Isothiocyanate Nicotinamide adenine dinucleotide phosphate Signal Transduction Food Science Biotechnology |
| Zdroj: | Molecular Nutrition & Food Research. 57:854-864 |
| ISSN: | 1613-4125 |
| DOI: | 10.1002/mnfr.201200689 |
| Popis: | Scope 6-Methylthiohexyl isothiocyanate (6-MTITC), one of the major bioactive ingredients in Japanese Wasabi, has revealed cytoprotective and cancer chemopreventive effects. This study aims to clarify the molecular mechanisms how 6-MTITC modulates nuclear factor E2-related factor 2 (Nrf2)/Kelchlike ECH-associating protein 1 (Keap1) system in antioxidant-responsive element (ARE)-mediated nicotinamide adenine dinucleotide phosphate (NADP): quinone oxidoreductase 1 (NQO1) expression. Methods and results HepG2 cells were treated with 6-MTITC with varying time and dose. NQO1, Nrf2, and Keap1 proteins were detected by Western blotting. ARE transactivation was detected by electrophilic mobility shift assay and reporter gene assay. Nuclear localization of Nrf2 was determined by immunocytochemistry assay. Ubiquitination of Nrf2 and Keap1 was detected using immunoprecipitation after treatment with MG132. Small interfering RNA was used to knockdown Nrf2 or Keap1. The results revealed that 6-MTITC modulated Nrf2/ARE pathway by stimulating Keap1 modification, and inhibiting Nrf2 ubiquitination and protein turnover. These actions finally increased nuclear Nrf2 accumulation and ARE-binding activity. Moreover, silencing Nrf2 markedly reduced ARE-driven activity induced by 6-MTITC. Conclusion 6-MTITC modulated ARE-driven NQO1 expression by stabilizing Nrf2 with enhanced Keap1 modification and decreased Nrf2 degradation. |
| Databáze: | OpenAIRE |
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