Diannexin attenuates release of MPs after hepatic IRI and decreases their pro-inflammatory and deleterious potential.

<p>MPs were obtained from mice pre-treated with 1 mg/kg iv Diannexin 5 min prior to 60 min liver IR and results compared with vehicle-treated/naïve mice subject to IR. <b>A</b>. FACS revealed significantly reduced resting endothelial cell (CD144), resting platelet (CD41) and activated platelet and endothelial cell (CD62P) subsets compared to naïve (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0104376#pone-0104376-g001" target="_blank">Fig.1C</a>). <b>Bi</b>. Serum ALT after 60 min ischemia and indicated reperfusion times in naïve and Diannexin-treated mice (n = 10 per cohort). * p<0.05 all experimental groups vs. sham. § p<0.05 Diannexin vs. 24 hr reperfusion. <b>Bii</b>. Platelets were isolated from sham-operated mice and exposed to 60 nM MPs (derived from mice subjected to 60 min ischemia, 2 hr reperfusion) in the absence or presence of 4 µg/ml Diannexin. FACS then performed using CD62P to quantify activated platelet subset as a proportion of total platelets. * p<0.05 experimental vs. control. # p<0.05 Diannexin vs. 60 nM MPs. <b>C, D</b>. Hepatocyte-specific ASPGR, E-selectin and pro-inflammatory VCAM, ICAM-1 protein expression diminished by Diannexin compared to naïve mice subjected to 60 min ischemia and indicated reperfusion times. + p<0.05 Diannexin vs. sham. & p<0.05 Diannexin vs. naïve. <b>E</b>. Pre-treatment of MPs with Diannexin 4 µg/ml inhibited neutrophil transmigration in ThinCert chambers. To ascertain effect of test compounds on transmigration potential, MPs were pre-treated with Diannexin (4 µg/ml) for 1 hr 37°C; thereafter, MPs were resuspended in PBS then recovered by prolonged centrifugation twice (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0104376#pone.0104376.s004" target="_blank">Materials and Methods S1</a> for detailed MP isolation protocol) prior to the transwell migration assay. * p<0.05 experimental vs. control. # p<0.05 Diannexin vs. 60 nM MPs.