br Materials and methods br Acknowledgements We thank

Materials and methods
Acknowledgements We thank Drs. Nathan Sherer (University of Wisconsin-Madison) and Bryan Cullen (Duke University) for generous reagent gifts. The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HIV-2ROD phage from Dr. Ronald Desrosiers. This work was supported in part by the Chinese government and NIH/NIDA DP1DA036463. R.E.S. is a NIDA HIV Avant Garde awardee.
Introduction Sepsis, a systemic inflammatory response caused by infection or injury, could lead to the development of tissue damage, septic shock, multiple organ dysfunction syndrome (MODS) and even death [1]. Despite significant advances in our understanding of the molecular and cellular mechanisms of sepsis, it is still one of the most important causes of death in clinical patients [2]. Current clinical therapies for sepsis are limited and include antibiotics, fluid resuscitation, and early goal-directed therapies (EGDT), which were recommended as guidelines by the International Surviving Sepsis Campaign [3]. The ongoing search for the pathogenesis of sepsis and novel therapeutic methods are highly urgent. Sepsis that occurs in overwhelming inflammatory responses to infections caused by pathogens or their products, such as lipopolysaccharide (LPS) plays an important role in this process. A key feature of the inflammatory response is the activation of pro-inflammatory cell, mainly macrophages and monocytes. During infection, LPS is released in the circulation to trigger pro-inflammatory Manumycin A australia to produce the early pro-inflammatory cytokines (e.g. tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), IL-6, interferon-γ (IFN-γ)) and the late mediator high mobility group box 1 (HMGB1) [4]. Once secreted outside the cell, these mediators could bind and active specific receptor and amplify the inflammatory reaction, leading to sever conditions such as systemic inflammatory response syndrome (SIRS) and MODS [2]. The excessive release of these mediators results in countless disturbances of the host immune system accompanied by tissue injury. Mechanistically, LPS acts on toll-like receptor 4 (TLR4) expressed on pro-inflammatory cells to induce the activation of two different signal pathways, nuclear factor-κB (NF-κB) signal pathway and mitogen-activated protein kinases (MAPKs), including extracellular signal-related kinase (ERK)-1/2, p38 MAPKs, or c-Jun NH2-terminal kinase (JNK) [5]. In previous studies, anti-inflammatory effects of pharmacological agents targeted at MAPKs, PI3K, ROS or NF-κB have been reported in animal models of inflammatory diseases [6]. Therefore, development of inhibitors targeted at different enzyme molecules of the LPS signaling cascades is an attractive strategy for the therapy of inflammatory diseases. KPT330 is a recently developed small-molecule inhibitor of the chromosome region maintenance 1 (CRM1) protein [7], which has been shown antitumor effects in leukemia, lung cancer and pancreatic cancer [8]. Previous studies showed that specific inhibiting of CRM1-mediated nuclear protein export, further increases the nuclear accumulation of IκBα and reduces TNF-α release [9]. However, the anti-inflammation effects of KPT330 treatment after sepsis induction and the molecular function of KPT330 on inflammatory response have not been evaluated. In this study, we further explored the potential effect of KPT330 in systemic inflammatory response in LPS-induced sepsis model in vivo and the molecular mechanism of anti-inflammation in macrophage in vitro.
Materials and methods
Results
Discussion The pathophysiology of sepsis is complex, which is a host\'s exaggerated immune response to severe infection increases mortality [13]. During the early phase, sepsis patients present with an exaggerated immune response, also described as a “cytokine storm”. And this acute hyperinflammation against infection leads to multiple organ dysfunction. It has been reported that a nuclear export factor CRM1 appears to be critical in assisting HMGB1 active export from the nucleus into the cytoplasm, which is a key “late-phase” inflammatory mediator of sepsis [14]. In this study, we evaluated the effects of KPT330, a potent and specific small molecule inhibitor of CRM1, in LPS-induced sepsis model in vivo and explored the underlying mechanism by performing a series of in vitro experients, and we found that KPT330 exhibited anti-inflammation effects and protection against sepsis.