Upregulation of NF-E2-related factor-2-dependent glutathione by carnosol provokes a cytoprotective response and enhances cell survival  

作　　者：Chien-chung CHEN Hui-ling CHEN Chia-wen HSIEH Yi-ling YANG Being-sun WUNG 

机构地区：[1]Department of Neurology, Saint Martin De Porres Hospital, Chiayi, Taiwan, China [2]Department of Biochemical Science and Technol- ogy, National Chiayi University, Chiayi, Taiwan, China [3]Department of Microbiology and Immunology, National Chiayi University, Chiayi, Taiwan, China

出　　处：《Acta Pharmacologica Sinica》2011年第1期62-69,共8页中国药理学报（英文版）

摘　　要：Aim： To explore whether glutathione （GSH） increased through Nrf-2 activation is involved in the cytoprotective effects of carnosol in HepG2 ceils. Methods： Human hepatoma cell line HepG2 were exposed to rosemarry essential oil or carnosol. Cell viability was measured using an Alamar blue assay. The production of intracellular GSH was determined using monochlorobimane. The level of protein or mRNA was examined by Western blotting or RT-PCR, respectively. Results： Rosemarry essential oil （0.005%-0.02%） and carnosol （5 and 10 mol/L） increased the intracellular GSH levels and GSH synthesis enzyme subunit GCLC/GCLM expression. Rosemary essential oil and carnosol increased nuclear accumulation of Nrf2 and enhanced Nrf2-antioxidant responsive element （ARE）-reporter activity. Transfection of the treated cells with an Nrf2 siRNA construct blocks GCLC/GCLM induction. Furthermore, pretreatment of the HepG2 cells with essential oil and carnosol exerted significant cyto- protective effects against H2O2 or alcohol. In TNFα-treated cells, the nuclear translocation and transcriptional activity of NF-KB was abolished for 12 h following carnosol pretreatment. Cotreatment with GSH also suppressed NF-KB nuclear translocation, whereas cotreatment with BSO, a GSH synthesis blocker, blocked the inhibitory effects of carnosol. Conclusion： This study demonstrated that Nrf2 is involved in the cytoprotective effects by carnasol, which were at least partially mediated through increased GSH biosynthesis.Aim： To explore whether glutathione （GSH） increased through Nrf-2 activation is involved in the cytoprotective effects of carnosol in HepG2 ceils. Methods： Human hepatoma cell line HepG2 were exposed to rosemarry essential oil or carnosol. Cell viability was measured using an Alamar blue assay. The production of intracellular GSH was determined using monochlorobimane. The level of protein or mRNA was examined by Western blotting or RT-PCR, respectively. Results： Rosemarry essential oil （0.005%-0.02%） and carnosol （5 and 10 mol/L） increased the intracellular GSH levels and GSH synthesis enzyme subunit GCLC/GCLM expression. Rosemary essential oil and carnosol increased nuclear accumulation of Nrf2 and enhanced Nrf2-antioxidant responsive element （ARE）-reporter activity. Transfection of the treated cells with an Nrf2 siRNA construct blocks GCLC/GCLM induction. Furthermore, pretreatment of the HepG2 cells with essential oil and carnosol exerted significant cyto- protective effects against H2O2 or alcohol. In TNFα-treated cells, the nuclear translocation and transcriptional activity of NF-KB was abolished for 12 h following carnosol pretreatment. Cotreatment with GSH also suppressed NF-KB nuclear translocation, whereas cotreatment with BSO, a GSH synthesis blocker, blocked the inhibitory effects of carnosol. Conclusion： This study demonstrated that Nrf2 is involved in the cytoprotective effects by carnasol, which were at least partially mediated through increased GSH biosynthesis.

关 键 词：CARNOSOL GLUTATHIONE Nrf2 NF-KB human hepatoma cell line HepG2 CYTOPROTECTION 

分 类 号：Q55[生物学—生物化学] Q26