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Physiol. Genomics 4: 137-145, 2000;
1094-8341/00 $5.00
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Received 8 August 2000; accepted in final form 12 October 2000.
Physiological Genomics 4:137-145 (2000)
1094-8341/00 $5.00 © 2000 American Physiological Society

Abrogation of thrombin-induced increase in pulmonary microvascular permeability in PAR-1 knockout mice

STEPHEN M. VOGEL1,*, XIAOPEI GAO1,*, DOLLY MEHTA1, RICHARD D. YE1, THERESA A. JOHN1, PATRICIA ANDRADE-GORDON2, CHINNASWAMY TIRUPPATHI1 and ASRAR B. MALIK1

1 Department of Pharmacology, University of Illinois College of Medicine, Chicago, Illinois 60612
2 The R. W. Johnson Pharmaceutical Research Institute, Spring House, Pennsylvania 19477

We investigated the function of proteinase-activated receptor-1 (PAR-1) in the regulation of pulmonary microvascular permeability in response to thrombin challenge using PAR-1 knockout mice (-/-). Lungs were isolated and perfused with albumin (5 g/100 ml)-Krebs solution at constant flow (2 ml/min). Lung wet weight and pulmonary artery pressure (Ppa) were continuously monitored. We determined the capillary filtration coefficient (Kfc) and 125I-labeled albumin (BSA) permeability-surface area product (PS) to assess changes in pulmonary microvessel permeability to liquid and albumin, respectively. Normal and PAR-1-null lung preparations received in the perfusate: 1) thrombin or 2) selective PAR-1 agonist peptide (TFLLRNPNDK-NH2). In control PAR-1 (+/+) mouse lungs, 125I-albumin PS and Kfc were significantly increased over baseline (by ~7- and 1.5-fold, respectively) within 20 min of {alpha}-thrombin (100 nM) challenge. PAR-1 agonist peptide (5 µM) gave similar results, whereas control peptide (5 µM; FTLLRNPNDK-NH2) was ineffective. At relatively high concentrations, thrombin (500 nM) or PAR-1 agonist peptide (10 µM) also induced increases in Ppa and lung wet weight. All effects of thrombin (100 or 500 nM) or PAR-1 agonist peptide (5 or 10 µM) were prevented in PAR-1-null lung preparations. Baseline measures of microvessel permeability and Ppa in the PAR-1-null preparations were indistinguishable from those in normal lungs. Moreover, PAR-1-null preparations gave normal vasoconstrictor response to thromboxane analog, U-46619 (100 nM). The results indicate that the PAR-1 receptor is critical in mediating the permeability-increasing and vasoconstrictor effects of thrombin in pulmonary microvessels.

proteinase-activated receptor; cultured endothelial cell monolayers; tethered ligand




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