Thrombomodulin domains attenuate atherosclerosis by inhibiting thrombin-induced endothelial cell activation

Autor: Yi-Heng Li, Guey Yueh Shi, Cheng Hsiang Kuo, Hua Lin Wu, Shu-Lin Liu, Hsi Ju Wei, Po Chiao Chang
Rok vydání: 2011
Předmět:
Neointima
Carotid Artery Diseases
Time Factors
Apolipoprotein B
Physiology
media_common.quotation_subject
Thrombomodulin
Vascular Cell Adhesion Molecule-1
Permeability
Mice
Thrombin
Apolipoproteins E
Physiology (medical)
medicine
Human Umbilical Vein Endothelial Cells
Animals
Humans
Protein Interaction Domains and Motifs
Receptor
PAR-1
Internalization
Blood Coagulation
Chemokine CCL2
media_common
Mice
Knockout
Binding Sites
biology
Chemistry
Cell adhesion molecule
Tumor Necrosis Factor-alpha
Monocyte
Atherosclerosis
Intercellular Adhesion Molecule-1
Molecular biology
Peptide Fragments
Recombinant Proteins
Endothelial stem cell
Mice
Inbred C57BL
Disease Models
Animal
medicine.anatomical_structure
Biochemistry
Mutation
biology.protein
Calcium
Cardiology and Cardiovascular Medicine
medicine.drug
Zdroj: Cardiovascular research. 92(2)
ISSN: 1755-3245
Popis: Aims Thrombin modulates the formation of atherosclerotic lesions by stimulating a variety of cellular effects through protease-activated receptor-1 (PAR-1) activation. Thrombomodulin (TM) inhibits thrombin effects by binding thrombin through its domains 2 and 3 (TMD23). We investigated whether recombinant TMD23 (rTMD23) could inhibit atherosclerosis via its thrombin-binding ability. Methods and results Wild-type mouse rTMD23 and three mutants with altered thrombin-binding sites, rTMD23 (I425A), rTMD23 (D424A/D426A), and rTMD23 (D424A/I425A/D426A), were expressed and purified in the Pichia pastoris expression system. Wild-type rTMD23 and rTMD23 (D424A/D426A) could effectively bind thrombin, activate protein C, and prolong thrombin clotting time, whereas rTMD23 (I425A) and rTMD23 (D424A/I425A/D426A) lost these functions. Wild-type rTMD23, but not rTMD23 (I425A), decreased both the thrombin-induced surface PAR-1 internalization and the increase in cytoplasmic Ca2+ concentrations in endothelial cells (ECs). Wild-type rTMD23 and rTMD23 (D424A/D426A) also inhibited thrombin-induced adhesion molecules and monocyte chemoattractant protein-1 expression and increased permeability in ECs, whereas rTMD23 (I425A) and rTMD23 (D424A/I425A/D426A) had no such effects. Furthermore, wild-type rTMD23 and rTMD23 (D424A/D426A) were effective in reducing carotid ligation-induced neointima formation in C57BL/6 mice and atherosclerotic lesion formation in apolipoprotein E-deficient (ApoE−/−) mice, whereas rTMD23 with the I425A mutation showed impairment of this function. Wild-type rTMD23, but not rTMD23 (I425A), also markedly suppressed the PAR-1, the adhesion molecules expression, and the macrophage content in the carotid ligation model and ApoE−/− mice. Conclusion rTMD23 protein significantly reduces atherosclerosis and neointima formation through its thrombin-binding ability.
Databáze: OpenAIRE
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