Modulation of monocyte adherence to endothelial cells by endothelin-1 involvement of Src (p60(src)) and JAK1-like kinases

L. J. Chisholm, P. S. Dovgan, Devendra K. Agrawal, P. E. McGregor, J. D. Edwards

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

Purpose: The purpose of this study was to determine the transmembrane signaling pathway by which endothelin-1 (ET-1) enhances monocyte adherence to human umbilical vein endothelial cells (HUVECs) and to investigate the role of tyrosine kinases in this mechanism. Methods: Adherence of purified human blood monocytes to HUVEC monolayers was assessed with radiolabeled monocytes. Tyrosine kinase activation was examined by immunoprecipitation and Western blotting. Results: ET-1 potentiated monocyte adherence to HUVECs in a biphasic manner with peaks at 10-10 mol/L and 10-7 mol/L. A potent antagonist to ET(B) receptors, when used alone, had no effect. However, the antagonist, when combined with ET-1, significantly enhanced monocyte adherence to HUVECs. Incubation of ET-1 (10-12 mol/L to 10-7 mol/L) with HUVECs activated tyrosine kinases in a biphasic manner as identified by immunoblotting with PY20 antibody to tyrosine phosphorylated proteins. Phosphorylated proteins with Mr 60, 110, and 130 kDa were observed after ET- 1 stimulation of HUVECs. Of interest, ET(A) or ET(B) receptor antagonists failed to antagonize the effect of ET-1. Rather, these receptor antagonists significantly augmented ET-1 induced tyrosine phosphorylation in HUVECs. Immunoprecipitation with antibodies to p60(src) and JAK1 kinases followed by immunoblotting with PY20 antibody suggested that ET-1 receptor-response coupling in HUVECs involves the activation of p60(src) and JAK1-like kinases. Conclusions: These data suggest an association between activation of p60(src) and JAK1-like kinases and monocyte adherence in response to ET-1. ET-1- induced monocyte adherence is upregulated by ET(B) receptor antagonist, suggesting a negative feedback on cell adhesion through this receptor.

Original languageEnglish
Pages (from-to)288-300
Number of pages13
JournalJournal of Vascular Surgery
Volume23
Issue number2
DOIs
Publication statusPublished - 1996
Externally publishedYes

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All Science Journal Classification (ASJC) codes

  • Cardiology and Cardiovascular Medicine
  • Surgery

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