The role of p38 mitogen-activated protein kinase/nuclear factor-ΚB transduction pathway on coagulation disorders due to endothelial injury induced by sepsis

Abstract

To determine the activation status of p38 mitogen-activated protein kinase (p38MAPK)/nuclear factor-ΚB (NF-ΚB) in coagulation disorders due to endothelial injury induced by sepsis. Human umbilical vein endothelial cells (HUVECs) were exposed to plasma obtained from 22 patients suffering from sepsis. Plasma was also obtained from 8 healthy individuals to serve as negative control, and tumor necrosis factor-α (TNF-α) was used as positive control. Phosphorylation and activity of p38MAPK and NF-ΚB were determined with enzyme-linked immunosorbent assay (ELISA), Western blotting, and immunofluorescence assay. The level of TNF-α (ng/L) in sepsis plasma was significantly higher than that in healthy plasma (155.68±89.74 vs. 5.00±0.47, P <0.01). Compared with healthy plasma in 20% concentration it was found when HUVECs were treated with sepsis plasma in 20% concentration, tissue factor (TF, μg/L) reached the peak at 180 minutes (5.87±0.14 vs. 1.25±0.11, P <0.01), von Willebrand factor (vWF, μg/L) reached the peak at 120 minutes (9.59±0.07 vs. 3.59±0.06, P <0.01), then they began to decline. When HUVECs were treated with sepsis plasma in 20% concentration increased phosphorylation and activity of p38MAPK and NF-ΚB, phosphorylation of p38MAPK occurred before phosphorylation of NF-ΚB (2 minutes vs. 5 minutes). When the inhibitor of p38MAPK (SB239063) was added, NF-ΚB phosphorylation (activation) and NF-ΚB nuclear translocation were inhibited. This study demonstrates that p38MAPK/NF-ΚB transduction pathway plays an important role in septic coagulopathy.