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 language | English |
|---|---|
| Pages (from-to) | 288-300 |
| Number of pages | 13 |
| Journal | Journal of Vascular Surgery |
| Volume | 23 |
| Issue number | 2 |
| DOIs | |
| State | Published - 1996 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Cardiology and Cardiovascular Medicine
- Surgery
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Modulation of monocyte adherence to endothelial cells by endothelin-1 involvement of Src (p60(src)) and JAK1-like kinases. / Chisholm, L. J.; Dovgan, P. S.; Agrawal, Devendra K.; McGregor, P. E.; Edwards, J. D.
In: Journal of Vascular Surgery, Vol. 23, No. 2, 1996, p. 288-300.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Modulation of monocyte adherence to endothelial cells by endothelin-1 involvement of Src (p60(src)) and JAK1-like kinases
AU - Chisholm, L. J.
AU - Dovgan, P. S.
AU - Agrawal, Devendra K.
AU - McGregor, P. E.
AU - Edwards, J. D.
PY - 1996
Y1 - 1996
N2 - 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.
AB - 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.
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U2 - 10.1016/S0741-5214(96)70273-2
DO - 10.1016/S0741-5214(96)70273-2
M3 - Article
VL - 23
SP - 288
EP - 300
JO - Journal of Vascular Surgery
JF - Journal of Vascular Surgery
SN - 0741-5214
IS - 2
ER -