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Molecular Biological Analysis of function and signal transduction of renin-binding protein

肾素结合蛋白功能和信号转导的分子生物学分析

基本信息

批准号：
09671631
负责人：
KANAYAMA Hiro-omi
金额：
$ 0.77万
依托单位：
The Universtiy of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

Renin binding protein (RnBP) is a protein that binds to renin to form a protein complex called high molecular weight (HMW) renin. This protein inhibits renin activity. In search of its physiological function gene expression of RnBP in rat aorta were examined by Northern blot hybridization. RnBP gene expression was observed in rat aorta, in addition to kidney, adrenal gland, brain, and ovary.Using In situ RT-PCR analysis to determine RnBP mRNA localization in rat kidney, it was suggested that RnBP mRNA was expressed in juxtamedullary glomerulus and vascular endothelial cells in glomerulus. The artcrial endothelial cells derived from porcine aorta exhibited high level of RnBP gene expression, detected by Northern blot hybridization analysis. In order to study renin gene expression in relation to that of RnBP, porcine renin cDNA fragment was isolated by RT-PCR analysis of preine kidney mRNA, followed by determination of the nucleotide sequence for porcine renin. Finally, RT- PCR analysis of porcine renin mRNA in endothelial cells has shown that renin gene is also expressed in the same endothelium. RnBP gene expression was shown to be up-regulated by Angiotensin II and TGF-beta, hut not by TNF-alfa in cultured endothelial cells.Therefore, RnBP regulation by angiotensn II suggests that RnBP participate in blood pressure regulation at vascular endothelum as a member of vascular renin-angiotensin system.

肾素结合蛋白（RnBP）是与肾素结合形成称为高分子量（HMW）肾素的蛋白质复合物的蛋白质。这种蛋白质抑制肾素活性。为探讨RnBP的生理功能，采用北方杂交技术检测了RnBP在大鼠主动脉中的基因表达。RnBP基因在大鼠主动脉、肾脏、肾上腺、脑和卵巢中均有表达，原位RT-PCR检测RnBP mRNA在大鼠肾脏中的定位结果表明，RnBP mRNA表达于肾小球髓质和血管内皮细胞。北方杂交结果显示，猪主动脉血管内皮细胞中RnBP基因表达水平较高。为了研究与RnBP基因表达的相关性，通过RT-PCR分析前肾mRNA来分离猪肾原酶cDNA片段，然后测定猪肾原酶的核苷酸序列。最后，对内皮细胞中的猪肾素mRNA进行RT-PCR分析，结果表明，在同一内皮细胞中也有肾素基因的表达。在培养的内皮细胞中，RnBP基因的表达受Ang Ⅱ和TGF-β的上调，而不受TNF-α的调节，提示RnBP可能参与血管内皮细胞的血压调节，是血管紧张素-血管紧张素系统的一员。