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GW23-e1868
TAP-SSL5 FUNCTIONS AS AN ANTICOAGULANT AND ANTI-INFLAMMATORY PROTEIN TO INHIBIT THROMBOSIS IN RAT AND ATTENUATE ATHEROSCLEROSIS IN APOE-KNOCKOUT MOUSE
  1. Xiaolong Qu1,
  2. Zhaofei Fang1,
  3. Min Li1,
  4. Dexing Huang2,
  5. Houyuan Hu1
  1. 1Department Cardiology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
  2. 2BakerIDI Heart & Diabetes Institute, Melbourne, Victoria, Australia

    Abstract

    Objectives This study aims to inhibit the inflammatory response and thrombosis in atherosclerotic lesions, by using the staphylococcal superantigen-like protein-5 (SSL5) to inhibit neutrophil activation, and tick anticoagulant peptide (TAP) to inhibit factor Xa (FXa) activation. To achieve this, we made a novel fusion protein TAP-SSL5 and tesed its dual anticoagulant and anti-inflammatory properties.t

    Methods We put six flexible amino acids sequence between TAP and SSL5 genes as a linker. The polycistronic fragment was constructed and then subcloned into a pET22b(+) expression vector. The binding ability of TAP-SSL5 to P-selectin glycoprotein ligand-1(PSGL-1) on leukocytes was verified by flow cytometry. The binding of TAP-SSL5 to PSGL-1 inhibited the adhesion of leukocytes to P-selectin-coated surface. FXa activity was determined by chromogenic substrate assay.

    Results We have observed that TAP-SSL5 inhibited FXa activity in a concentration-dependent manner and TAP-SSL5 reduced ferric chloride-induced thrombosis in the inferior vena cava of rat. In a high-fat diet induced atherosclerotic model by using ApoE-knockout (ApoE−/−) mouse, male ApoE−/− mice were grouped to receive intraperitoneal treatment with either TAP-SSL5 (3 mg/kg/day), SSL5 (2 mg/kg/day) or vehicle separately. After 12 weeks of the respective treatment, we have observed that TAP-SSL5 reduced the atherosclerotic plague formation by 48% compared to controls. We also found TAP-SSL5 could down-regulate several kinds of inflammatory cytokine expressions in vascular wall which were detected by mouse inflammatory antibody array.

    Conclusions TAP-SSL5 fusion protein has promising anti-inflammatory and anti-thrombosis properties, and it acquires the potential for the prevention of atherosclerotic lesion and thrombus formation.

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