title: Heparin regulates transcription of endothelin-1 gene in endothelial cells.
abstract: 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.
authors.affiliation: Department of Cardiology, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Kazue
authors.initials: K
authors.lastname: Kuwahara-Watanabe
authors.forename: Chiaki
authors.initials: C
authors.lastname: Hidai
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ikeda
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Aoka
authors.forename: Ken-ichiro
authors.initials: K
authors.lastname: Ichikawa
authors.forename: Nobuo
authors.initials: N
authors.lastname: Iguchi
authors.forename: Masako
authors.initials: M
authors.lastname: Okada-Ohno
authors.forename: Jinko
authors.initials: J
authors.lastname: Yokota
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kasanuki
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Kawana
mesh_terms: 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*
chemical_list: 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