Figure 5.

OxPAPC enhances endothelial barrier function and attenuates agonist-induced endothelial barrier dysfunction in vitro and in vivo. (A) HPAECs grown on coverslips were treated with OxPAPC (15 µg/mL, 30 minutes) and dual immunofluorescence staining with Texas-Red phalloidin and VE-cadherin was performed to monitor actin cytoskeleton and cell junction remodeling. The confocal images illustrated the barrier enhancing effects of OxPAPC as evidenced by increased F-actin and VE-Cadherin staining. (B) HPAECs grown on biotinylated gelatin substrate were pre-treated with OxPAPC (15 µg/mL, 30 minutes) followed by thrombin stimulation (0.5 Units/mL, 10 minutes) and cell permeability was visualized using FITC-avidin as a tracer. OxPAPC treatment protected thrombin-induced permeability as demonstrated by reduced green fluorescence that depicts areas permeable for FITC-avidin. (C) Intravenous injection of (1.5 mg/kg) of OxPAPC after 5 hours of LPS instillation (0.7 mg/kg, intratracheal) in mice protects against LPS-induced lung injury as detected by Evans blue staining of lung.