Oxidative Stress, Hypertension and an FGF-binding Protein

氧化应激、高血压和 FGF 结合蛋白

• 批准号: 7218285
• 负责人: Anton Wellstein
• 金额: $ 33.47万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7218285
• 关键词: AIDS-Associated Nephropathy; Acute Kidney Failure; Address; Adult; Angiotensin II; Animals; Binding; Binding Proteins; Biochemical; Blood Pressure; Blood Vessels; Cell Survival; Cells; Chemical Injury; Circadian Rhythms; Complement; Cultured Cells; Development; Endothelium; Epithelial; Epithelial Cells; Epithelium; Extracellular Matrix; Fibroblast Growth Factor; Fibroblast Growth Factor 1; Fibroblast Growth Factor 2; Fibroblast Growth Factor Receptors; Figs - dietary; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; Gene Expression; Generations; Human; Hypertension; Immunoprecipitation; In Vitro; Injury; Kidney; Knockout Mice; MAPK14 gene; MAPK8 gene; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mesenchymal; Mitogen-Activated Protein Kinases; Modeling; Monitor; Mus; Normal tissue morphology; Organ; Oxidative Stress; Pathway interactions; Patients; Proteins; Reactive Oxygen Species; Reading; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Regulation; Research Design; Research Personnel; Role; Sampling; Series; Signal Transduction; Signal Transduction Alteration; Signaling Protein; Skin; Smooth Muscle Myocytes; Stress; Superoxide Dismutase; Superoxides; Tetracycline; Tetracycline Control; Tetracyclines; Tissues; Transgenes; Transgenic Mice; Tubular formation; Two-Dimensional Gel Electrophoresis; Up-Regulation; Work; Wound Healing; blood pressure regulation; cell type; day; experimental analysis; extracellular; in vivo; mimetics; permanent cell line; programs; receptor; repaired; research study; response; role model; selective expression; tempol; transgene expression

Fibroblast growth factors (FGF-1 or -2) are present at significant concentrations in most normal tissues in the adult. However, these FGFs are immobilized in an inactive state on the extracellular matrix and it is only poorly understood how they are solubilized and activated to reach their extracellular receptors. One mechanism through which FGFs can be mobilized is by binding to secreted binding proteins (BPs) and we showed that BP1 can enhance the activity of locally stored, immobilized FGFs. BP1 expression is controlled by stress pathways in cultured cells and found upregulated after wounding, toxic or infectious injury of the skin or kidneys. BP1 expression in mice carrying an inducible BP1 transgene caused a significant rise in mean arterial blood pressure (MAP) by +30 mm Hg within two days of transgene induction and analysis of vascular contractility showed a sensitization to angiotensin II. The rise of MAP after BP1 transgene expression was inhibited by systemic administration of the superoxide dismutase mimetic Tempol suggesting an essential role of oxidative stress. We hypothesize that BP1/FGF signaling modulates the sensitivity of blood vessels towards contractile signaling and propose to study this under the following aims: Aim 1. To evaluate the contribution of FGF-2 or other FGFs to the BP1-induced hypertensive effect. We will study blood pressure, vessel contractility and renal tubular function in FGF-2(-/-) mice that are crossed with mice carrying an inducible BP1 transgene. Systemic administration of BP1 and FGF-2 will complement this. Aim 2. To study the contribution of kidney expression of BP1 to blood pressure regulation, vessel contractility and renal tubular function we will use mice harboring a HoxB7-controlled, tetracycline inducible BP1 transgene. To evaluate the role of endogenous BP1 to oxidative stress-regulated blood pressure, we will generate mice that are null for BP1 expression. Aim 3. To study the intracellular cross-talk between BP1 / FGF signaling and G-protein coupled receptor pathways we will monitor signal transduction and phenotypic effects in preglomular smooth muscle cells from experimental animals. Biochemical signaling via known integrators of the pathways (i.e. MAPKs) and proliferation/cell survival and superoxide generation will be used as read-outs. Mass spectrometry to identify new signaling proteins in the cross-talk will complement this.

成纤维细胞生长因子（FGF-1或-2）在大多数正常组织中以显著浓度存在， 成年人然而，这些FGF以非活性状态固定在细胞外基质上，并且其仅在细胞外基质中存在。 对它们如何溶解和激活以到达其细胞外受体知之甚少。一 FGF的动员机制是通过与分泌的结合蛋白（BP）结合， 显示BP 1可以增强局部储存的固定化FGF的活性。BP 1表达受控制 通过培养细胞中的应激途径，发现在创伤、毒性或感染性损伤后上调， 皮肤或肾脏。在携带诱导型BP 1转基因的小鼠中，BP 1的表达引起了BP 1基因表达的显著增加。 平均动脉血压（MAP）在转基因诱导的两天内升高+30 mm Hg， 血管收缩性显示对血管紧张素II敏感。转BP 1基因后MAP升高 通过全身给予超氧化物歧化酶模拟物Tempol抑制表达 这表明氧化应激的重要作用。我们假设BP 1/FGF信号转导调节了 血管对收缩信号的敏感性，并建议在以下目标下对此进行研究： 目标1。评估FGF-2或其他FGF对BP 1诱导的高血压效应的贡献。我们将 研究FGF-2（-/-）小鼠的血压、血管收缩性和肾小管功能， 携带可诱导BP 1转基因的小鼠。BP 1和FGF-2的全身给药将对此进行补充。 目标二。为了研究BP 1在肾脏中的表达对血压调节的作用， 收缩性和肾小管功能，我们将使用携带HoxB 7控制的四环素诱导的 BP 1转基因。为了评估内源性BP 1在氧化应激调节血压中的作用，我们 将产生BP 1表达无效的小鼠。 目标3.研究BP 1/ FGF信号通路与G蛋白偶联受体之间的细胞内串扰 我们将监测肾小球前体平滑肌细胞的信号转导和表型效应 从实验动物身上。通过已知的途径整合子（即MAPK）的生化信号传导， 增殖/细胞存活和超氧化物生成将被用作读出。质谱鉴定 新的信号传导蛋白质将补充这一点。