Urokinase-type plasminogen activator (uPA) induces pulmonary microvascular endothelial permeability through low density lipoprotein receptor-related protein (LRP)-dependent activation of endothelial nitric-oxide synthase.

Abstract

Urokinase plasminogen activator (uPA) and PA inhibitor type 1 (PAI-1) are elevated in acute lung injury, which is characterized by a loss of endothelial barrier function and the development of pulmonary edema. Two-chain uPA and uPA-PAI-1 complexes (1-20 nM) increased the permeability of monolayers of human pulmonary microvascular endothelial cells (PMVECs) in vitro and lung permeability in vivo. The effects of uPA-PAI-1 were abrogated by the nitric-oxide synthase (NOS) inhibitor L-NAME (N(D)-nitro-L-arginine methyl ester). Two-chain uPA (1-20 nM) and uPA-PAI-1 induced phosphorylation of endothelial NOS-Ser(1177) in PMVECs, which was followed by generation of NO and the nitrosylation and dissociation of β-catenin from VE-cadherin. uPA-induced phosphorylation of eNOS was decreased by anti-low density lipoprotein receptor-related protein-1 (LRP) antibody and an LRP antagonist, receptor-associated protein (RAP), and when binding to the uPA receptor was blocked by the isolated growth factor-like domain of uPA. uPA-induced phosphorylation of eNOS was also inhibited by the protein kinase A (PKA) inhibitor, myristoylated PKI, but was not dependent on PI3K-Akt signaling. LRP blockade and inhibition of PKA prevented uPA- and uPA-PAI-1-induced permeability of PMVEC monolayers in vitro and uPA-induced lung permeability in vivo. These studies identify a novel pathway involved in regulating PMVEC permeability and suggest the utility of uPA-based approaches that attenuate untoward permeability following acute lung injury while preserving its salutary effects on fibrinolysis and airway remodeling.

• Authors: Anastasia M Makarova, Tatiana V Lebedeva, Taher Nassar, Abd Al-Roof Higazi, Jing Xue, Maria E Carinato, Khalil Bdeir, Douglas B Cines, Victoria Stepanova
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 286 Issue 26 Pg. 23044-53 (Jul 01 2011) ISSN: 1083-351X [Electronic] United States
• PMID: 21540184 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Enzyme Inhibitors
• Low Density Lipoprotein Receptor-Related Protein-1
• Plasminogen Activator Inhibitor 1
• SERPINE1 protein, human
• Serpin E2
• Serpine2 protein, mouse
• Nitric Oxide Synthase Type III
• Nos3 protein, mouse
• Cyclic AMP-Dependent Protein Kinases
• Urokinase-Type Plasminogen Activator
• NG-Nitroarginine Methyl Ester

Topics

• Acute Lung Injury (genetics, metabolism, pathology)
• Animals
• Blood-Air Barrier (metabolism, pathology)
• Capillary Permeability (drug effects, genetics)
• Cell Line
• Cyclic AMP-Dependent Protein Kinases (antagonists & inhibitors, genetics, metabolism)
• Enzyme Inhibitors (pharmacology)
• Fibrinolysis (drug effects, genetics)
• Humans
• Low Density Lipoprotein Receptor-Related Protein-1 (genetics, metabolism)
• Mice
• Mice, Knockout
• NG-Nitroarginine Methyl Ester (pharmacology)
• Nitric Oxide Synthase Type III (antagonists & inhibitors, genetics, metabolism)
• Phosphorylation (drug effects, genetics)
• Plasminogen Activator Inhibitor 1 (genetics, metabolism, pharmacology)
• Pulmonary Edema (genetics, metabolism, pathology)
• Respiratory Mucosa (metabolism, pathology)
• Serpin E2 (genetics, metabolism, pharmacology)
• Urokinase-Type Plasminogen Activator (genetics, metabolism, pharmacology)