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采用实时逆转录聚合酶链反应对大鼠内皮素-1及血管活性肠收缩肽/内皮素-2基因表达进行定量分析。

Quantitative analysis of endothelin-1 and vasoactive intestinal contractor/endothelin-2 gene expression in rats by real-time reverse transcriptase polymerase chain reaction.

作者信息

Uchide T, Adur J, Fukamachi T, Saida K

机构信息

Biosignaling Department, National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology, Tsukuba, Ibaraki, Japan.

出版信息

J Cardiovasc Pharmacol. 2000 Nov;36(5 Suppl 1):S5-8. doi: 10.1097/00005344-200036051-00004.

Abstract

We established a real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) system for the analysis of rat endothelin-1 (ET-1) and vasoactive intestinal contractor (VIC)/ET-2 gene expression. We used this technique to examine the expression levels in rat in 16 different organs. ET-1 gene expression was observed in all organs examined, while VIC mRNA was detected in some organs such as heart, lung, ovary, stomach, and intestine. Ovary and intestine express both ET-1 and VIC mRNA at high levels, suggesting the importance of both peptides in these organs. In addition, we examined the gene expression levels in intestinal epithelial and mesenchymal tissues from rat fetuses at 16.5 and 19.5 days postcoitus (E16.5 and E19.5). We observed distinct differences in the temporal gene expression patterns for ET-1 and VIC in fetal intestinal epithelial tissue. In fetal mesenchymal tissue the expression level of ET-1 is significantly higher than that of VIC, and the levels of both genes remain unchanged over the time period observed. These findings suggest distinct biological roles and gene regulation mechanisms for ET-1 and VIC in intestinal epithelial and mesenchymal tissues.

摘要

我们建立了一种实时定量逆转录聚合酶链反应(RT-PCR)系统,用于分析大鼠内皮素-1(ET-1)和血管活性肠收缩肽(VIC)/内皮素-2(ET-2)基因的表达。我们运用该技术检测了大鼠16种不同器官中的表达水平。在所检测的所有器官中均观察到ET-1基因的表达,而在心脏、肺、卵巢、胃和肠等部分器官中检测到了VIC mRNA。卵巢和肠中ET-1和VIC mRNA均高水平表达,提示这两种肽在这些器官中的重要性。此外,我们检测了妊娠16.5天和19.5天(E16.5和E19.5)大鼠胎儿的肠上皮和间充质组织中的基因表达水平。我们观察到胎儿肠上皮组织中ET-1和VIC的基因表达模式在时间上存在明显差异。在胎儿间充质组织中,ET-1的表达水平显著高于VIC,并且在观察期内这两种基因的水平均保持不变。这些发现提示ET-1和VIC在肠上皮和间充质组织中具有不同的生物学作用和基因调控机制。

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