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Fig. 6 | Respiratory Research

Fig. 6

From: Unfractionated heparin ameliorates pulmonary microvascular endothelial barrier dysfunction via microtubule stabilization in acute lung injury

Fig. 6

Effect of UFH or the p38 MAPK inhibitor SB203580 pretreatment on LPS-induced increase in endothelial cell permeability and F-actin remodeling. HPMECs were pretreated with UFH (10 U/ml) or SB203580 (10 μM) for 30 min, followed by LPS (10 μg/ml) stimulation for 6 h. a The TEER across the cells were measured. Both UFH and SB203580 protected cells from LPS-induced endothelial barrier dysfunction by increasing TEER of the HPMEC monolayer. b The influx of FITC-conjugated dextran across the cells was measured. Both UFH and SB203580 protected cells from LPS-induced endothelial barrier dysfunction by decreasing FITC-conjugated dextran leakage. c Actin cytoskeletal remodeling and area of inter-endothelial gaps. Actin cytoskeletal remodeling was examined by immunofluorescence staining with TRITC-phalloidin. Arrows indicate intercellular gaps. UFH or SB203580 pretreatment decreased LPS-induced formation of stress fibers (scale bars = 50 μm). The area of gaps in the microscope fields was assessed and normalized to the area of gaps induced with LPS alone. UFH and SB203580 prevented paracellular gaps induced by LPS. Nine microscope fields from three parallel experiments were analyzed for each group. d Western blot analysis was used to evaluate the levels of GEF-H1 and MYPT1 phosphorylation. Equal protein loading was confirmed using GAPDH. LPS-induced GEF-H1 expression and MYPT1 phosphorylation were attenuated by UFH as well as SB203580. Data are representative of three independent experiments. t-test, * P < 0.05 compared to the vehicle group, t-test, # P < 0.05 compared to the LPS-treated group

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