Thus, the present study was investigated to provide possible mechanisms that CA treatment offers against em t /em -BHP-induced oxidative stress in liver cells. MAPKs and Nrf2 activation had not been previously investigated. Thus, the present study was investigated to provide possible mechanisms that CA treatment offers against em t /em -BHP-induced oxidative stress in liver cells. In addition, it is well worth mentioning that em t /em -BHP was used as an oxidative agent with this study. Because em t /em -BHP is not relevant to human being LY2334737 exposure, it may be appropriate to test other oxidative stress agents to human being that may be LY2334737 exposed to humans for future experiments. To survive under a variety of environmental stresses, hepatocytes maintain a cellular defense systems that shields them against oxidative difficulties [25, 26]. One of these system requires phase II drug-metabolizing enzymes, such as glutathione-S-transferase and UDP-glucuronosyltransferase [27], and antioxidant enzymes, such as HO-1, NADP(H):quinone oxidoreductase-1 (NQO-1), and GCL [28, 29]. Our earlier study reported that CA treatment only increased only GCL catalytic subunit, GCLC mRNA level in normal phase cell [4]. However, as can be evinced from the data in the present study, cell treatment with CA led to a dose-dependent significant increase in the manifestation of not only GCLC but also GCLM, compared with cells treated only with em t /em -BHP. These discrepancies LY2334737 may be due to the concentration of CA treated in the cells, and/or the incubation time treated in the CA in the presence or absence of em t /em -BHP. In the previous experiment [4], HepG2 cells were treated having a concentration of CA from 62?M up to 250?M for 8?h without em t /em -BHP treatment, whereas the maximum concentration of CA used in this experiment was 20?M for 24?h followed by em t /em -BHP treatment for 2?h. On the other hand, the L-02 liver cells which were incubated with CA LY2334737 (10 and 50?M) for 15?min, and then incubated with 7.5?mM acetaminophen for 48?h had no effect on GCLC and GCLM mRNA/protein [30]. Huang et al. reported that up-regulated the mRNA/protein manifestation of GCLC and GCLM was observed in rat main hepatocytes treated with flavones including 25?M chrysin and apigenin for 24?h [31]. Treatment of Natural264.7 cells with em t /em -BHP significantly reduced GCLC and GCLM mRNA levels, and treatment of these cells with 25?M licochalcone A, a natural phenol for 18?h, led to the recovery of both GCLC and GCLM gene manifestation levels [32]. Our results exhibited that cytotoxicity caused by em t /em -BHP-induced oxidative stress was recovered by CA treatment by way of the up-regulation of the expression of detoxifying enzymes like HO-1, GCLC, and GCLM. These enzyme-encoding genes, whose expression is associated with detoxification activity, were regulated by a consensus em cis /em -element located at the 5-flanking promoter region, such as the antioxidant response element (ARE) [33]. The transcription factor Nrf2 plays a key role in the antioxidant redox cycle associated with cell survival, because it is an essential component of the ARE-binding transcription factor [8]. Investigating Nrf2 translocation, we observed that cells treated with CA experienced a significant and dose-dependent nuclear accumulation of Nrf2. On the other hand, in cells treated with CA was observed a reduction in the amount of cytosolic Nrf2 compared with cells treated with em t /em -BHP alone. Previously, various studies demonstrated that candidate materials of chemopreventive brokers can lead to the Nrf2 accumulation in nucleus and promoting of Nrf2-dependent gene expression [10, 34]. The change in the redox caused by oxidative stress is known to alter many signaling pathways, including MAPKs [35]. MAPK pathways mediated by ERK, JNK, and p38 have been demonstrated to play a central role in transducing extracellular signals to the nucleus [36]. Results from a study exhibited that short-term Rabbit Polyclonal to C56D2 treatment of rat prostate endothelial cells with em t /em -BHP increased the level of p38 and ERK phosphorylation [37]. However, our result showed.