Upregulation of NF-E2-related factor-2-dependent glutathione by carnosol provokes a cytoprotective response and enhances cell survival
Upregulation of NF-E2-related factor-2-dependent glutathione by carnosol provokes a cytoprotective response and enhances cell survival
复制标题
DOI：
10.1038/aps.2010.181
复制
发表时间：
2011-01-01
影响因子：
8.2
通讯作者：
Wung, Being-sun
中科院分区：
文献类型：
--
作者：
Chen, Chien-chung;Chen, Hui-ling;Wung, Being-sun
目的：探讨通过NRF-2激活而增加的谷胱甘肽(GSH)是否参与了卡诺醇对人肝癌细胞株HepG2的细胞保护作用。用阿拉玛蓝比色法测定细胞存活率。用一氯二苯甲烷测定细胞内GSH的产生。结果：迷迭香精油(0.005%~0.02%)和卡诺索尔(5和10moL/L)均能增加细胞内GSH水平和GSH合成酶亚单位Gclc/Gclm的表达。迷迭香精油和卡诺醇增加了Nrf2的核积累，并增强了Nrf2-抗氧化反应元件(ARE)-报告活性。用Nrf2 siRNA构建的处理细胞可阻断GCLC/GCLM的诱导。此外，用精油和卡诺醇对HepG2细胞进行预处理，对过氧化氢和乙醇具有明显的细胞保护作用。在经肿瘤坏死因子α处理的细胞中，肌醇预处理12h后，核转位和核转录活性被取消。与GSH共同处理也可抑制核转录因子-kappaB的核转位，而与GSH合成阻断剂BSO共同处理可阻断卡诺醇的抑制作用。结论：Nrf2参与了卡诺醇的细胞保护作用，这种保护作用至少部分是通过增加GSH的生物合成来实现的。
Aim: To explore whether glutathione (GSH) increased through Nrf-2 activation is involved in the cytoprotective effects of carnosol in HepG2 cells.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 cytoprotective effects against H2O2 or alcohol. In TNF alpha-treated cells, the nuclear translocation and transcriptional activity of NF-kappa B was abolished for 12 h following carnosol pretreatment. Cotreatment with GSH also suppressed NF-kappa B 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.