It has been proved that the transcription of MnSOD is regulated by STAT319, 29 depending on the JAK2/STAT3 protein complex

that is formed by the interaction of Hes proteins with STAT3.30 We found that Hes5 was the major downstream effector of Notch signaling in hepatocytes during I/R injury. Thus, disruption of Notch signaling resulted in decreased Hes5-STAT3 complex, and overexpressing constitutively active STAT3 or Hes5 rescued MnSOD expression, leading to reduced ROS levels and hepatocyte apoptosis subjected to I/R in the absence of Notch signaling. In summary, the data presented in this study learn more establish a signal axis by which canonical Notch signaling regulates hepatic I/R injury: the activated Notch receptors up-regulate Hes5 through transcription factor RBP-J, and Hes5 facilitates STAT3 activation through the formation of a Hes5-JAK2/STAT3 complex,30 which in turn activates the transcription of MnSOD gene to scavenge ROS and restricts I/R injury (Supporting Fig. 12, left). The JAK/STAT pathway mediates cytokine signaling and participates

in the initiation, propagation, and resolution of inflammation.32 The basic selleckchem players in this pathway include four JAK kinases and seven STAT members, with other modifiers such as SOCSs. The JAK2/STAT3-SOCS3 module mediates proinflammation or anti-inflammation signaling, depending on cell types and other environmental cues, and is involved in both hepatic and myocardial I/R injury.33, 34 We have shown recently that Notch signaling down-regulates JAK2/STAT3 signaling through the up-regulation of SOCS3 in macrophages.23 This signal could be enhanced through the auto-amplification of Notch signaling by TLR-induced and RBP-J–dependent induction of Notch ligands,35 likely to result in down-regulation of MnSOD and increased ROS levels, to facilitate the destruction of ingested pathogens in macrophages G protein-coupled receptor kinase (Supporting Fig. 12, right). In tissue parenchymal cells such as hepatocytes, as shown in the current study, I/R also up-regulates

Notch signal activation, but it assists JAK2/STAT3 signaling without the activation of SOCS3 expression, resulting in the up-regulation of MnSOD and increased scavenging of ROS, restricting the extent of tissue damage. Therefore, it appears that, as one of the early response signals, the Notch pathway plays differential roles through JAK2/STAT3-MnSOD in macrophages and hepatocytes—namely, increasing ROS in macrophages to destroy pathogens but reducing ROS in hepatocytes to protect cells. This scenario might also be an explanation of the contradictory observations about the roles of Notch signaling in myocardial and brain I/R injuries.12, 14 However, mechanisms such as epigenetic elements by which Notch signal differentially regulates SOCS3 expression between macrophages and hepatocytes should be determined by further studies. We thank Klaus Rajewski for Mx-Cre transgenic mice, J. C. Zúñiga-Pflücke for OP9 derivatives, and Yongzhan Nie for plasmids expressing STAT3 mutants.