REGULATION OF RENIN AND PREECLAMPTIC HYPERTENSION

肾素和子痫前期高血压的调节

基本信息

批准号：
7217183
负责人：
Dinesh Manilal Shah
金额：
$ 14.72万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-04-19 至 2006-03-31
项目状态：
已结题

项目摘要

DESCRIPTION: (Adapted from the Investigator's Abstract) Many tissues secrete an inactive pro-form of renin. A 43 amino acid pro- segment removal to produce active renin is believed to occur, primarily in the kidney. Recent data on transgenic mouse model of preeclampsia support a role of reproductive renin in its pathogenesis and our data on increased decidual renin expression in human preeclampsia confirms this role. For renin to have such a functional role, cellular processing to active renin should occur locally. The cellular and molecular aspects of the uterine renin- angiotensin system have not been adequately investigated. Presence of estrogen and progesterone response elements in the promoter of renin gene suggests a role for the sex steroids in the regulation of prorenin expression. Our recent data suggest that progesterone treatment of endometrial stromal cells (in vitro decidualization) increases active renin secretion by increasing prorenin expression and by increasing conversion of prorenin to renin in a hormone- specific manner, and that decidual cells from human gestation process prorenin to renin. Our most recent co-transfection experiment in HEK293 cells and cathepsin B inhibition in decidual cells suggest that cathepsin B may function as a prorenin convertase. We hypothesize that pathologic increase in decidual renin may proximately initiate preeclampsia, and sex steroids may induce expression of prorenin and its proconvertase (PC), thereby regulating mature renin secretion. Therefore, the objectives are: 1) To demonstrate that decidual stromal cells in vitro secrete active mature renin, as assayed by activity and by N terminal sequencing; 2) To identify the decidual cell endoprotease responsible for prorenin activation by (i) northern analysis with probes for PCs and cathepsin B, and (ii) defining constitutive versus regulated renin secretion, and (iii) examining the effect of cathepsin B inhibition on prorenin processing; (b) To examine regulation of endoprotease expression; 3) To confirm that the candidate-activating enzyme is the specific prorenin convertase by (a) immuno-microscopic co-localization with prorenin in the vesicles and (b) functional verification by co-transfection of prorenin and candidate-activating enzyme expression vectors in a standard cell line and demonstration of processing to mature active renin. These studies will improve our understanding of the role of progesterone in the regulation of reproductive renin and may provide a novel direction for investigating the pathophysiology of preeclampsia.

描述：（改编自研究者的摘要）许多组织分泌了肾素的不活跃形式。据信主要在肾脏中，据信将去除43个氨基酸的促进段以产生活性肾素。先兆子痫的转基因小鼠模型的最新数据支持生殖肾素在其发病机理中的作用，以及我们在人类前启示识中增加骨素表达增加的数据证实了这一作用。为了使肾素具有这种功能性作用，应在本地进行细胞加工至活动肾素。子宫肾上腺素 - 血管紧张素系统的细胞和分子方面尚未得到充分研究。肾素基因启动子中雌激素和孕酮反应元件的存在表明性类固醇在调节prorenin表达中的作用。我们最近的数据表明，子宫内膜基质细胞的孕酮治疗（体外脱义化）通过增加肾上腺素的表达并增加了肾上腺素向肾素的转化来增加活性肾素分泌，并以激素特异性的方式以及从人类妊娠过程中的骨化细胞从肾蛋白到肾蛋白中的cant骨细胞。我们最近在HEK293细胞和结论细胞中抑制组织蛋白酶B的共染色实验表明组织蛋白酶B可能起到prorenin转化酶的作用。我们假设骨肾上数的病理增加可能近距离启动子痫，并且性类固醇可能诱导prorenin及其前旋转酶（PC）的表达，从而调节成熟的肾素分泌。因此，目的是：1）证明，根据活性和n末端测序分析的分泌活性成熟的肾素的决结质细胞分泌活性成熟肾素； 2）通过（i）通过（i）使用PC和组织蛋白酶B探针进行北方分析来鉴定负责prorenin激活的决明细胞内蛋白酶，以及（ii）定义组成型与调节的肾素分泌与调节的肾素分泌，以及（iii）检查calter蛋白B抑制对prorenin处理的影响；（b）检查内切蛋白酶表达的调节； 3）确认候选激活酶是（a）通过（a）在囊泡中与prorenin的免疫显微镜共定位的特异性prorenin转化酶，（b）通过共转染预侵了prorenin和候选酶表达在标准的细胞线上和演示rention intication in Activation retivation in Activation in Activation in Activation in Activation in Activation in Activation in Activation in Activation intivation norkitions in Activation norkity norkitions in native intivation norkiations的功能验证。这些研究将提高我们对孕激素在生殖肾素调节中的作用的理解，并可能为研究前酮的病理生理学提供新的方向。