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

Hsi Ju Wei, Yi Heng Li, Guey Yueh Shi, Shu Lin Liu, Po Chiao Chang, Cheng Hsiang Kuo, Hua Lin Wu

研究成果: Article同行評審

32 引文 斯高帕斯(Scopus)

摘要

AimsThrombin 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 resultsWild-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 Ca 2 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.ConclusionrTMD23 protein significantly reduces atherosclerosis and neointima formation through its thrombin-binding ability.

原文English
頁(從 - 到)317-327
頁數11
期刊Cardiovascular Research
92
發行號2
DOIs
出版狀態Published - 2011 十一月 1

All Science Journal Classification (ASJC) codes

  • 生理學
  • 心臟病學與心血管醫學
  • 生理學(醫學)

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