Transcription factor decoy to study the molecular mechanism of negative regulation of renin gene expression in the liver in vivo.
Transcription factor decoy to study the molecular mechanism of negative regulation of renin gene expression in the liver in vivo.
复制标题
转录因子诱饵研究体内肝脏肾素基因表达负调控的分子机制。
DOI：
10.1161/01.res.84.9.1059
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发表时间：
1999
影响因子：
20.1
通讯作者：
Horiuchi,M
中科院分区：
文献类型：
--
作者：
Tomita,S;Tomita,N;Yamada,T;Zhang,L;Kaneda,Y;Morishita,R;Ogihara,T;Dzau,VJ;Horiuchi,M
- 在肾脏的肾小球细胞中大量合成肾素，但在肝脏中很少或不合成。我们前期的体外和生化研究表明，小鼠肾素基因的组织特异性表达受其5′侧翼区负调控元件（NRE）与NRE结合蛋白（NREB）的特异性相互作用的调控。在这项研究中，我们研究的假设，这种相互作用之间的NRE在启动子区的大鼠肾素基因和NREB在肝脏中的抑制在体内该组织中的肾素基因表达。我们使用NRE转录因子诱饵（TFD）双链寡核苷酸通过门静脉输注体内转染到大鼠肝脏中。凝胶迁移率变动分析表明，转染的NRE TFD阻断了内源性NREB与大鼠肾原酶基因的结合。这导致增强的肝脏肾素mRNA的表达，免疫组织化学检测的肝脏中的肾素，因此，增加血浆肾素浓度。总之，这些结果证明了NREB在体内抑制大鼠肝脏中的肾素基因表达的重要性，并表明了通过TFD方法进行体内肾素基因调节的可能性。
—Renin is synthesized in high quantities in the juxtaglomerular cells of the kidney, but little or none is synthesized in the liver. Our previous in vitro and biochemical studies have demonstrated that tissue-specific expression of the mouse renin gene is regulated by the specific interaction between negative regulatory element (NRE) in the 5′-flanking region of the renin gene and NRE binding protein (NREB). In this study, we examined the hypothesis that this interaction between the NRE in the promoter region of the rat renin gene and the NREB in the liver contributes to the suppressed renin gene expression in this tissue in vivo. We used in vivo transfection of NRE transcription factor decoy (TFD) double-stranded oligonucleotide into the rat liver via portal vein infusion. A gel mobility shift assay showed that transfected NRE TFD blocked endogenous NREB binding with the rat renin gene. This resulted in enhanced hepatic renin mRNA expression, immunohistochemical detection of renin in the liver, and consequently, increased plasma renin concentration. Taken together, these results document the importance of NREB in the inhibition of renin gene expression in rat liver in vivo and suggest the possibility of in vivo renin gene modulation by the TFD approach.