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Constitutive and Inducible Systems for Genetic In Vivo Modification of Mouse Hepatocytes Using Hydrodynamic Tail Vein Injection
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Mutagenesis studies toward understanding the intracellular signaling mechanism of antithrombin

J-S Bae1, A R Rezaie

  • 1Edward A Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO 63104, USA.

Journal of Thrombosis and Haemostasis : JTH
|March 27, 2009
PubMed
Abstract

Insights

Antithrombin (AT) has anti-inflammatory effects mediated by syndecan-4 and G-protein coupled receptors. A modified AT (AT/Proth-2) shows protective signaling without anticoagulant activity, suggesting safer anti-inflammatory drug potential.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Pharmacology

Background:

  • Antithrombin (AT) exhibits anti-inflammatory and antiangiogenic properties.
  • Investigating the intracellular signaling mechanisms of AT is crucial for understanding its biological functions.

Purpose of the Study:

  • To elucidate the intracellular signaling pathways of AT.
  • To analyze the roles of wild-type and mutant serpins in cellular signaling.
  • To differentiate between anticoagulant and anti-inflammatory activities of AT.

Main Methods:

  • Utilized wild-type and mutant serpins (RCL and heparin-binding site mutants).
  • Assessed cellular effects in LPS-stimulated endothelial cells using permeability and neutrophil adhesion assays.
  • Investigated the involvement of pertussis toxin (PTX), syndecan-4, sphingosine 1-phosphate receptor 1 (S1P(1)), prostacyclin, and NF-kappaB.

Main Results:

  • Wild-type AT and RCL mutant AT/Proth-2 demonstrated potent barrier protection and inhibited neutrophil adhesion via NF-kappaB inhibition.
  • Heparin-binding site mutants (AT-K114E, AT-K125E) lacked protective activity; AT-K114E showed pro-apoptotic effects.
  • Syndecan-4 and PTX were essential for AT's protective effect, while S1P(1) was not involved.

Conclusions:

  • AT's protective effect is prostacyclin-dependent, mediated by syndecan-4 and a PTX-sensitive G-protein coupled receptor.
  • The RCL mutant AT/Proth-2 retains protective signaling but lacks anticoagulant activity, offering potential as a safer anti-inflammatory therapeutic.

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