p38 MAP kinase-dependent regulation of endothelial cell permeability.
We have previously shown that thrombin induces endothelial cell barrier dysfunction via cytoskeleton activation and contraction and have determined the important role of endothelial cell myosin light chain kinase (MLCK) in this process. In the present study we explored p38 MAP kinase as a potentially important enzyme in thrombin-mediated endothelial cell contractile response and permeability. Thrombin induces significant p38 MAP kinase activation in a time-dependent manner with maximal effect at 30 min, which correlates with increased phosphorylation of actin- and myosin-binding protein, caldesmon. Both SB-203580 and dominant negative p38 adenoviral vector significantly attenuated thrombin-induced declines in transendothelial electrical resistance. Consistent with these data SB-203580 decreased actin stress fiber formation produced by thrombin in endothelium. In addition, dominant negative p38 had no effect on thrombin-induced myosin light chain diphosphorylation. Thrombin-induced total and site-specific caldesmon phosphorylation (Ser789) as well as dissociation of caldesmon-myosin complex were attenuated by SB-203580 pretreatment. These results suggest the involvement of p38 MAP kinase activities and caldesmon phosphorylation in the MLCK-independent regulation of thrombin-induced endothelial cell permeability.
Division of Pulmonary and Critical Care Medicine, Johns Hopkins University, School of Medicine, Baltimore, Maryland 21224, USA.
Talaibek
T
Borbiev
Anna
A
Birukova
Feng
F
Liu
Saule
S
Nurmukhambetova
William T
WT
Gerthoffer
Joe G N
JG
Garcia
Alexander D
AD
Verin
D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002199:Capillary Permeability / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000201:enzymology; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology*; D004789:Enzyme Activation / Q000187:drug effects; D006490:Hemostatics / Q000494:pharmacology; D009218:Myosins / Q000378:metabolism; D010766:Phosphorylation; D011651:Pulmonary Artery / Q000166:cytology*; D022502:Stress Fibers / Q000187:drug effects / Q000378:metabolism; D013917:Thrombin / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
D002148:Calmodulin-Binding Proteins; D006490:Hemostatics; D048051:p38 Mitogen-Activated Protein Kinases; D013917:Thrombin; D009218:Myosins