KALLIKREIN-RELATED ENZYMES IN CARDIOVASCULAR REGULATION

心血管调节中激肽释放酶相关酶

• 批准号: 5213425
• 负责人: A G SCICLI
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5213425
• 关键词: Baculoviridae; Xenopus oocyte; angiotensin /renin /aldosterone hypertension; blood pressure; disease /disorder model; enzyme inhibitors; gene expression; glomerular filtration rate; hormone regulation /control mechanism; in situ hybridization; kallikreins; kidney function; laboratory rabbit; laboratory rat; monoclonal antibody; polymerase chain reaction; protein kinase C; recombinant proteins; renal glomerulus; renal hypertension; spontaneous hypertensive rat; thrombin; ultrasound blood flow measurement; vascular resistance; vasoconstrictors

We have recently discovered a new protein which exhibits potent vasoconstrictor activity when applied to isolated vascular rings. We demonstrated that it is an enzyme belonging to the kallikrein family of serine proteases. The gene for this protein has been identified and the nucleotide sequence of its mRNA is available. We named the substance submandibular enzymatic vasoconstrictor (SEV) because it was isolated from the rat submandibular gland and requires enzymatic activity to contract arterial rings. We found that the mRNA for SEV is present in arteries, cultured vascular smooth muscle cells and kidney. When injected into rats, SEV caused a marked increase in blood pressure. We propose to test the general hypothesis that SEV participates in the regulation of vascular resistance and renal function. Furthermore, an unknown member of the kallikrein family is present in the vascular pole of the glomerulus, where it may play a role in the regulation of glomerular function. To further our understanding of the physio pathological role of these new kallikrein- related enzymes, we propose: I): to localize SEV and its mRNA in tissue using the polymerase chain reaction, in situ hybridization, immunohistochemistry and RIA. In addition, we will determine whether vascular tissue contains binding sites for SEV and whether SEV releases a vasoconstrictor peptide(s); II): to determine: a) whether SEV is released as an endocrine and/or exocrine hormone. We will study release of SEV into the salivary and urinary compartments of the submandibular gland and kidney, and in the venous effluent of these organs, respectively, b) the factor(s) that regulate tissue concentration and release of SEV and mRNA levels. III): to determine the effects of acute administration of SEV on blood pressure, cardiac output and renal function, and whether blockade of endogenous SEV with Fab fragments of monoclonal antibodies alters any of these parameters. IV): To identify, purify and characterize the kallikrein-related protein expressed in the vascular pole of the glomerulus.

我们最近发现了一种新的蛋白质，它具有强大的功效 当应用于孤立的血管环时具有血管收缩活性。 我们 证明它是一种属于激肽释放酶家族的酶 丝氨酸蛋白酶。 该蛋白质的基因已被鉴定，并且 其mRNA的核苷酸序列是可用的。 我们将这种物质命名为 颌下酶促血管收缩剂 (SEV)，因为它是从 大鼠颌下腺，需要酶活性来收缩 动脉环。 我们发现 SEV 的 mRNA 存在于动脉中， 培养血管平滑肌细胞和肾脏。 当注射到老鼠体内时， SEV 导致血压显着升高。 我们建议测试 SEV 参与血管调节的一般假设 抵抗力和肾功能。 此外，一名身份不明的成员 激肽释放酶家族存在于肾小球的血管极，其中 它可能在肾小球功能的调节中发挥作用。 为了进一步 我们对这些新的激肽释放酶的生理病理作用的理解- 相关酶，我们建议：I)：在组织中定位SEV及其mRNA 使用聚合酶链式反应、原位杂交、 免疫组织化学和放射免疫分析。 此外，我们将确定是否 血管组织含有 SEV 的结合位点以及 SEV 是否释放 血管收缩肽； II): 确定: a) 是否释放SEV 作为内分泌和/或外分泌激素。 我们将研究将 SEV 释放到 下颌下腺的唾液室和泌尿室 肾脏，以及这些器官的静脉流出物中，分别，b) 调节 SEV 和 mRNA 的组织浓度和释放的因子 水平。 III): 确定 SEV 急性给药对 血压、心输出量和肾功能，以及是否阻断 带有单克隆抗体 Fab 片段的内源性 SEV 会改变以下任何一项： 这些参数。 IV): 鉴定、纯化和表征 激肽释放酶相关蛋白在血管极表达 肾小球。