アデノシンによる血管内皮細胞でのエンドセリン1(ET-1)遺伝子発現の抑制について

概要

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Endothelin-1 (ET-1) may have a role in the development of reperfusion injury, and adenosine has been reported to have important functions in the development of ischemic preconditioning. The aim of the present study was to investigate the influence of adenosine on ET-1 gene expression in endothelial cells. Total RNA from cultured bovine aortic endothelial cells (BAEC) was extracted and hybridized with ^<32>P-labeled bovine ET-1 CDNA probes. To determine the transcriptional activity of the ET-1 promoter, we used plasmids including -1.7kb and -204bp upstream of the transcription initiation site, respectively. These were transfected to BAEC, and subsequently, RNase protection assay was performed. To evaluate the effect of adenosine on the protein binding activity at AP-1 and GATA sites of the ET-1 promoter, we performed gel shift assay. ET-1 mRNA expression was maximally suppressed by adenosine at 90 min. Adenosine significantly decreased transcriptional activity of the ET-1 promoter, suggesting that the adenosine reactive site is within the -204 bp of ET-1 promoter. Gel mobility shift assay showed that the AP-1 binding activities were suppressed. The AP-1 site of the ET-1 promoter might be one of the adenosine responsive elements.
東京女子医科大学の論文
1996-11-25