BRADYKININ RECEPTORS IN HYPERTENSION

这项修订提案的目的是澄清以下表述 缓激肽B2和B受体在血压中的作用 和心血管调节。激肽是一种有效的血管扩张肽 由激肽释放酶从前体激肽原中释放出来。Kinins绑定 BK受体并介导一系列广泛的生物学效应 包括血管调节、离子转运、平滑肌收缩和 放松、疼痛和发炎。有两种类型的激活素 受体B2和B1在生物学特性和原发方面的不同 结构。这个项目是基于这样一种假设，即BK 受体在血压调节中的作用是被支持的 根据我们令人鼓舞的初步结果，B2的反义抑制 转基因大鼠体内受体引起血压升高 过度表达人类B2受体的小鼠会出现低血压。自.以来 上一次提交的申请，我们继续取得进展 BK受体的分子克隆、表达和定位 开发了必要的生化和分子探针，并建立了 构成当前应用程序基础的技术。这个 这项提议的具体目的是：(L)克隆和刻画BK 来自人类和大鼠的受体基因，并创建DNA构建 使用和不使用腺病毒载体进行体细胞基因传递和 转基因小鼠发育，(2)分析调控因素 α-细胞培养体系中BK受体基因转录的研究 缺失分析、定点突变、凝胶滞留和 足迹分析，以及通过直接基因进行体内缺失分析 传递，(3)分析BK受体在中枢和中枢神经中的作用 反义抑制和顺铂对血压的外周调节 正常血压和高血压大鼠的联合基因传递，(4) 高表达BK的转基因小鼠的建立和鉴定 受体基因和分析血液等表型变化 血压变化、肾功能和平滑肌收缩。这个 拟议的研究可以提供关于 BK受体的结构、调节和功能 治疗高血压的治疗药物设计， 心血管疾病和炎症。

The objective of this revised proposal is to elucidate the expression and role of bradykiritin (BK) B2 and B receptors in blood pressure and cardiovascular regulation. Kinins are potent vasodilating peptides that are released from precursor kininogens by kallikreins. Kinins bind to BK receptors and mediate a broad spectrum of biological effects including vasodllation, ion transport, smooth muscle contraction and relaxation, pain and inflammation. There are two types of kinin receptors, B2 and B1 which differ in biological properties and primary structures. This project is based on the hypothesis that the BK receptor plays a role in blood pressure regulation which is supported by our encouraging preliminary results that antisense inhibition of B2 receptors in rats causes an increase of blood pressure while transgenic mice over-expressing human B2 receptors are hypotensive. Since the last submission of this application, we continue to make progress in molecular cloning, expression and locallzation of BK receptors, developed essential biochemical and molecular probes and established technologies which form the basis for the current application. The specific aims of this proposal are: (l) to clone and characterize the BK receptor genes from human and rat, and to create DNA constructs with and without adenoviral vectors for somatic gene delivery and transgenic mouse development, (2) to analyze elements which regulate the BK receptor genes' transcription in a-cell culture system by deletion analysis, site-directed mutagensis, gel retardation and footprinting assays, and in vivo by deletion analysis via direct gene delivery, (3) to analyze the role of BK receptors in central and peripheral regulation of blood pressure by antisense inhibition and by combination gene delivery in normotensive and hypertensive rats, (4) to develop and characterize transgenic mice over-expressing BK receptor genes and to analyze phenotypic alterations such as blood pressure changes, renal function and smooth muscle contraction. The proposed studies could provide fundamental information about the structure, regulation and function of BK receptors in order to facilitate the design of therapeutic agents for treating hypertension, cardiovascular disease and inflammation.