Efficacy of AGP in both endotoxemia and CLP support the potential utility of this novel, natural colloidal resuscitation fluid. The ability of AGP to maintain liver perfusion and decrease leukocyte adherence to the liver microvasculature could arise from numerous previously suggested AUY-922 manufacturer potential mechanisms, ranging from altering the ratio

of pro-inflammatory to anti-inflammatory cytokines and signals in hepatic inflammation, to restoring glycocalyx/barrier functions of the liver microcirculation, to directly binding and sequestering LPS. Of these possibilities, we selected the last one for further investigation, given that it was amenable to testing using methodologies already employed in this study. When AGP was combined with LPS and then injected intraperitoneally, it attenuated the pro-inflammatory effects of LPS on the hepatic microcirculation, at least with respect to leukocyte adhesion to PSV and sinusoidal perfusion. AGP has been shown to bind to LPS in two in vitro studies [25, 16]. If AGP bound LPS in the peritoneal space, it may have prevented the endotoxin from reaching the circulation and exerting systemic

effects, given the slow uptake of AGP from the peritoneal space into the circulation detected in our clearance experiments with radiolabeled AGP. Alternatively, LPS and AGP may not have interacted in the peritoneal space, but instead both reached selleck chemicals the circulation, where AGP exerted the anti-inflammatory effects we previously observed. To discriminate more fully between these

possibilities, we amended our experimental endotoxemia protocol to permit administration of both AGP and LPS intravenously, by reducing the LPS dose to 0.08 mg/kg, avoiding the mortality likely to ensue from an intravascular 5 mg/kg LPS dose. While administration of AGP just prior to LPS injection into the vasculature resulted in a non-significant trend toward decreased inflammation, pre-incubation of AGP with LPS significantly improved liver perfusion and reduced leukocyte adherence in both the post-sinusoidal venules and the sinusoids. Although in hindsight the latter experiment was likely underpowered, taken together our results support the concept that AGP is an LPS-binding ADAMTS5 protein and demonstrate this binding can have consequences in vivo. The anti-inflammatory effects of AGP manifested in the hepatic microcirculation are consistent with previous reports that infusion of pharmacological quantities of AGP purified from healthy cattle or humans limited mortality in disease models of uncontrolled inflammation [15, 20, 26]. However, they differ from two reports suggesting that AGP mediates a failure of leukocyte migration to the site of infection, both in normal and diabetic mice subjected to the CLP procedure. Mestriner et al. found that human AGP administered at the remarkably low dose of 4 μg/rat (approximately 0.