Isolation and characterization of a heparin with low antithrombin activity from the body of Styela plicata (Chordata-Tunicata). Distinct effects on venous and arterial models of thrombosis

Abstract 

Introduction

A heparin preparation with low antithrombin activity and different disaccharide composition than mammalian heparin was isolated from the body of the ascidian Styela plicata (Chordata-Tunicata). The disaccharide composition and the effect of the invertebrate glycan on venous and arterial models of thrombosis was investigated.

Methods and results

High performance liquid chromatography of the products formed by a mixture of heparin lyases showed that the ascidian heparin is composed mainly by ΔUA(2SO₄)-1→4-β-d-GlcN(SO4) (47.5%), ΔUA(2SO₄)-1→4-β-d-GlcN(SO₄)(6SO₄) (38.3%) disaccharides and smaller amounts of the disaccharides ΔUA(2SO₄)-1→4-β-d-GlcN(SO₄)(3SO₄)(6SO₄) (2.8%) and ΔUA(2SO₄)-1→4-β-d-GlcN(SO₄)(3SO₄) (8.0%). The invertebrate heparin has an aPTT activity of 18 IU/mg and an antithrombin-mediated antithrombin and anti-factor Xa activities 10-fold lower than that of mammalian heparin. In a venous model of thrombosis in the vena cava, S. plicata heparin inhibits only 80% of thrombosis at a dose 10-fold higher than that of the mammalian heparin that inhibits 100% of thrombosis. However, in an arterio-shunt model of arterial thrombosis, both S. plicata and mammalian heparin possess equivalent antithrombotic activities. It is also shown that at equivalent doses, ascidian heparin has a lower bleeding effect than mammalian heparin.

Conclusion

The antithrombin-mediated anticoagulant activity of heparin polymers is not directly related to antithrombotic potency in the arterio-venous shunt. The results of the present work suggest that heparin preparations obtained from the body of S. plicata may have a safer therapeutic action in the treatment of arterial thrombosis than mammalian heparin.

Abbreviations:: ΔUA, α-Δ^4,5 unsaturated hexuronic acid, ΔUA(2SO₄), α-Δ^4,5 unsaturated hexuronic acid 2-sulfate, α-l-IdoA, α-l-iduronic acid, α-l-IdoA-2(SO₄), α-l-iduronic acid 2-sulfate, GlcN(SO₄), GlcN(SO₄)(6SO₄) and GlcN(SO4)(3SO₄)(6SO₄), derivatives of d-glucosamine, bearing a sulfate ester at position N, and at both positions N and 6, and at positions N, 3 and 6, respectively, GlcNAc, N-acetyl-d-glucosamine, GlcNAc(6SO₄), N-acetyl-d-glucosamine 6-sulfate, HPLC, high-performance liquid chromatography, FPLC, fast protein liquid chromatography.

Keywords: Heparin, Dermatan sulfate, Antithrombotic effect, Arterio-venous model, Venous model, Ascidian