Deletion-mapping of the cis-regulating element responsible for elevated expression of the ET-1 gene in SHR VSMC

• Main Authors: Su Wei-Shuo, 蘇偉碩
• Other Authors: Chao Chung-Faye
• Format: Others
• Language: en_US
• Published: 1999
• Online Access: http://ndltd.ncl.edu.tw/handle/75079713181225349898

碩士 === 國防醫學院 === 生物及解剖學研究所 === 87 === 英文摘要 As a strong vasoconstrictor in vivo, the role of endothelin-1(ET-1) in the pathogenesis of primary hypertension has been well documented. According to our previous study, ET-1 secreted from spontaneously hypertensive rats (SHR) aortic smooth muscle cell was about 2.5 times higher than age matched Wistar-Kyoto (WKY) rats. For evaluation the different expression of ET-1 in vascular smooth muscle cell between SHR and WKY rats, four ET-1 promoter construct plasmids conjugated with luciferase reporter gene were constructed by different deletion mutant construction from a 1.3kb SD rat ET-1 promoter, followed with transfection luciferase assaya were performed. 1.3 kb SD rat ET-1 promoter sequence was revealed about 85% homology to the human ET-1 promoter sequence. Sequence analysis from rat ET-1 promoter showed that there are several consensus sequences for cis-acting elements, such as AP-1, GATA-2, calcium responsive element (CaRe), and GHF-1, located in this promoter region. It was indicated from our results that 1.3 bp ET-1 promoter expression in cultured vascular smooth muscle cells of SHR presented about 2 times higher than in that of WKY rats. When the ET-1 promotor reduced from 1.3kb to 1.1kb, the expression of luciferase reporter activity in SHR and WKY VSMC were similar. It is suggested that there are some cis-regulating elements which between 1.3-1.1 kb of ET-1 promotor were responsible for higher ET-1 promoter activity in SHR VSMC compared to WKY rat. According to the sequence analysis, a GHF-1 consensus sequence is located at position -1289 to -1283 of this 166 bp sequence. Does GHF-1 is responsible for the different expression of ET-1 in SHR and WKY rat VSMC need further investigation.