THROMBIN INDUCES THE PREPROENDOTHELIN-1 GENE IN ENDOTHELIAL-CELLS BY A PROTEIN-TYROSINE KINASE-LINKED MECHANISM

被引:44
作者
MARSEN, TA
SIMONSON, MS
DUNN, MJ
机构
[1] UNIV CLEVELAND HOSP,DEPT MED,DIV NEPHROL,CLEVELAND,OH 44106
[2] CASE WESTERN RESERVE UNIV,SCH MED,DEPT MED,CLEVELAND,OH 44106
[3] CASE WESTERN RESERVE UNIV,SCH MED,DEPT PHYSIOL & BIOPHYS,CLEVELAND,OH 44106
关键词
THROMBIN; ENDOTHELIN; ENDOTHELIAL CELLS; PROTEIN TYROSINE KINASE; PROTEIN KINASE C;
D O I
10.1161/01.RES.76.6.987
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Thrombin stimulates synthesis and secretion of endothelin-1 (ET-1), a vasoactive peptide that triggers responses in the vascular endothelium and smooth muscle. We investigated the signal transduction pathways by which thrombin stimulates preproET-1 gene expression and ET-1 peptide secretion in macrovascular cells (human umbilical vein endothelial cells [HUVECs] and bovine pulmonary artery endothelial cells [BPAECs]) and microvascular cells (human microvascular endothelial cell line [HMEC-1]). Thrombin (4 U/mL) stimulated maximal induction of ET-I peptide secretion and preproET-1 mRNA after 2 hours in HUVECs and BPAECs and after 1 hour in HMEC-1. A synthetic thrombin receptor activator peptide confirmed ligand-specific receptor actions to induce preproET-1 mRNA. Protein kinase C (PKC) activation by phorbol ester transiently induced preproET-1 mRNA but had no effect on ET-1 peptide synthesis. PKC inhibitors sangivamycin and calphostin C and PKC depletion failed to suppress thrombin-stimulated preproET-1 mRNA. Adenylate cyclase and cAMP-dependent protein kinase did not participate in thrombin-induced preproET-1 gene activation. Thrombin stimulated a rapid increase in phosphotyrosine-containing proteins, suggesting a role for tyrosine phosphorylation in thrombin signaling. These data demonstrate that thrombin induces the preproET-1 gene and ET-1 peptide synthesis by a PKC-independent PTK-dependent pathway in macrovascular and microvascular endothelial cells. Protein tyrosine kinase inhibitors herbimycin A and genistein blocked thrombin-stimulated preproET-1 mRNA and peptide secretion, whereas daidzein, which lacks inhibitory activity, did not suppress thrombin-induced ET-1.
引用
收藏
页码:987 / 995
页数:9
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