Heparin regulates transcription of endothelin-1 gene in endothelial cells.
Heparin, which is widely used as an anticoagulant, has been shown to have antiatherosclerotic and antihypertensive effects in animals and humans. These effects are mediated by the inhibition of endothelin-1 (ET-1) production in endothelial cells. To clarify the mechanism of this inhibition, we investigated the effect of heparin on transcriptional regulation of the ET-1 gene in bovine aortic endothelial cells (BAEC) cultured in fetal calf serum. ET-1 mRNA expression was significantly suppressed by heparin in a dose-dependent manner. Promoter analysis revealed that the minimum ET-1 promoter containing only the GATA and AP-1 sequences as positive cis-acting sites in the ET-1 promoter is sufficient for this suppression. Gel mobility shift assays using oligonucleotides encoding the ET-1 AP-1 and ET-1 GATA sites confirmed that both AP-1 and GATA binding activities in BAEC nuclear extract were markedly inhibited by heparin. Western blot analyses indicated that heparin completely blocked extracellular signal-regulated kinase (ERK) activation, and inhibiting ERK activity resulted in loss of heparin-dependent inhibition of the ET-1 gene. These data indicate that the ET-1 mRNA level is negatively regulated by heparin at the transcription level, through modification of AP-1 and GATA protein binding activities, which direct the ET-1 promoter in BAEC. This effect may be mediated, at least in part, through inhibition of ERK activity.
Kuwahara-Watanabe
Kazue
K
Department of Cardiology, Tokyo Women's Medical University, Tokyo, Japan.
Hidai
Chiaki
C
Ikeda
Hiromi
H
Aoka
Yoshikazu
Y
Ichikawa
Ken-ichiro
K
Iguchi
Nobuo
N
Okada-Ohno
Masako
M
Yokota
Jinko
J
Kasanuki
Hiroshi
H
Kawana
Masatoshi
M
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology*; D001011:Aorta / Q000166:cytology; D002417:Cattle; D002478:Cells, Cultured; D015536:Down-Regulation; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism*; D019332:Endothelin-1 / Q000235:genetics*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology*; D010766:Phosphorylation / Q000187:drug effects; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*
D000925:Anticoagulants; D019332:Endothelin-1; D005419:Flavonoids; D012333:RNA, Messenger; D018808:Transcription Factor AP-1; D014157:Transcription Factors; D006493:Heparin; D048049:Extracellular Signal-Regulated MAP Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one