Nrf2 then heterodimerizes with small Maf and binds to ARE, eventually resulting in transcriptional activation Lenalidomide manufacturer of the ARE mediated metabolizingdetoxifying and antioxi dant genes. We report in this study that digitofla vone strongly induced Nrf2 protein expression and nucleus accumulation. The rapid accu mulation of Nrf2 in the nucleus Inhibitors,Modulators,Libraries in response to digitofla vone is consistent with reported results with other Nrf2 activators, such as PEITC and celecoxib, and with the Nrf2 degradation Inhibitors,Modulators,Libraries inhibitors such as eckol. The Nrf2ARE pathway activates approximately 100 cytoprotective genes. In this study, digitoflavone ele vated the mRNA and protein levels of several ARE mediated antioxidantdetoxifying genes in Caco 2 cells.

Knockdown of Nrf2 by Nrf2 targeted siRNA markedly suppressed the digitoflavone induced GCSc, GCSm expression, suggesting that digitoflavone up regulates Nrf2 dependent activation of the ARE regulated genes. Nrf2 controls the expression of GCSc and GCSm, which together catalyze the rate limiting step in GSH biosynthesis. Involvement of GSH in the digitoflavone Inhibitors,Modulators,Libraries induced cytoprotection against oxidative injury could not be excluded, because increasing GSH levels would be expected to reduce ROS levels and antagonize the ROS induced cell death. In this study, treatment of cells with digitoflavone resulted in decreased H2O2 induced oxidative stress, and cell death. Activation of Nrf2 involves regulation of protein kinases, which may induce Nrf2 phosphorylation and nuclear translocation. The MAPK cascade, PI3KAKT, and PKC signaling pathways have been reported to influ ence the Nrf2ARE pathway.

For example, phosphor ylation of Nrf2 by PKC promotes its release from Keap1 and inhibition of PI3K attenuates the nuclear trans location Inhibitors,Modulators,Libraries of Nrf2 and transcription of ARE mediated genes. To identify which signal cascade controlled activa tion of Nrf2 by digitoflavone, we Inhibitors,Modulators,Libraries examined the effects of PI3K inhibitor, ERK12 inhibitor, and p38 MAPK inhibitor on the digitoflavone induced Nrf2 up regulation. Our re sults demonstrated that PI3KAKT and ERK12 are not involved in the digitoflavone induced activation of the Nrf2ARE pathway because their inhibitor had no effect on enhanced digitoflavone induced Nrf2 up regulation. On the contrary, inhibition of p38 MAPK by SB202190 leads to decrease of the digitoflavone induced Nrf2 up regulation, indicating that the digitoflavone induced Nrf2 activation is dependent on the activation of p38 MAPKs. Inhibition of p38 also abrogated the digitoflavone induced translocation of Nrf2 to nucleus and the antioxidant defense effect, demonstrating that the crucial role of p38 in the Nrf2 dependent activation of ARE and suggesting Volasertib that Nrf2 is a downstream effector of p38 kinase in response to digitoflavone treatment.