Hypertension and Collagen: Effect of Ac-SDKP

高血压和胶原蛋白：Ac-SDKP 的作用

• 批准号: 7904241
• 负责人: NOUR-EDDINE RHALEB
• 金额: $ 36.25万
• 依托单位: HENRY FORD HEALTH SYSTEM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7904241
• 关键词: Affinity; Amino Acid Sequence; Amino Acids; Angiotensin II; Angiotensin-Converting Enzyme Inhibitors; Angiotensins; Anti-Inflammatory Agents; Anti-inflammatory; Aorta; Attenuated; Binding; Binding Proteins; Binding Sites; Biochemical; Blood Pressure; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Cardiac; Cardiovascular system; Cell Proliferation; Cell membrane; Cells; Chinese Hamster Ovary Cell; Clathrin-Coated Vesicles; Cloning; Code; Collagen; Complementary DNA; Coronary artery; DTR gene; Dactinomycin; Diabetes Mellitus; Drosophila genus; Dynamin; EGF gene; ELK1 gene; Endocytosis; Endothelin; Endothelin-1; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Experimental Models; Extracellular Matrix; Extracellular Signal Regulated Kinases; Fibroblasts; Fibrosis; Gene Combinations; Heart failure; Heparin Binding; Human; Hypertension; Hypertrophy; In Vitro; Infiltration; Inflammation; Ionophores; Kidney; Kidney Diseases; Kinins; Label; Lead; Left ventricular structure; Link; MAP Kinase Gene; MAPK phosphatase; Macrophage Activation; Mass Spectrum Analysis; Matrix Metalloproteinases; Mediating; Mediator of activation protein; Membrane; Mitogen-Activated Protein Kinases; Mitogens; Molecular Weight; N-terminal; Names; Okadaic Acid; Organ; PTPN6 gene; Peptides; Peptidyl-Dipeptidase A; Phosphorylation; Phosphotransferases; Physiology; Plasma; Production; Protein Kinase C; Protein Tyrosine Phosphatase; Proteins; Proteomics; RNA Interference; Radio; Rattus; Receptor Protein-Tyrosine Kinases; Regulation; Renal function; Renin-Angiotensin-Aldosterone System; Research; Research Personnel; Small Interfering RNA; Src homology 2 domain-containing, transforming protein 1; System; Techniques; Technology; Testing; Tissues; Transactivation; Transforming Growth Factor beta; Vanadates; Xenopus oocyte; analog; cDNA Library; cardiovascular risk factor; goralatide; improved; inhibitor/antagonist; macrophage; monocyte; phosphatase inhibitor; phosphatase-1 kinase; prevent; programs; receptor; receptor coupling; receptor downregulation; receptor internalization; response; src-Family Kinases; tool; transcription factor

DESCRIPTION (provided by applicant): Hypertension is a major risk factor for .cardiovascular and renal diseases. Inflammation and components of the extracellular matrix (EM) have a negative impact on the physiology and function of end target organs such as the arteries, heart and kidneys in hypertension. Blocking angiotensin-converting enzyme (ACE) decreases angiotensin II (Ang II) and increases kinins, leading to decreased cardiovascular inflammation, hypertrophy and collagen. ACE inhibitors (ACEi) increase plasma Ac-SDKP, a negative regulator of cell proliferation present in plasma and tissue. In hypertension and heart failure, Ac-SDKP prevents monocyte/macrophage infiltration and fibrosis in the aorta, kidneys and left ventricle (LV). By virtue of its anti- fibrotic and anti-inflammatory effects, Ac-SDKP was able to improve renal function in hypertension, diabetes and other experimental models of renal diseases. However, the mechanism(s) or receptor(s) involved in Ac- SDKP's cardiovascular and renal effects are not fully understood. We hypothesize that Ac-SDKP exerts its anti-inflammatory and anti-fibrotic effects on the cardiovascular and renal systems in hypertension via specific receptor(s) located on the plasma membrane, contributing to end organ protection. In aim I we will identify and characterize Ac-SDKP receptors using pharmacological tools [ l]Hpp-Aca-SDKP, 5(6)FAM-SDKP and new analogues of Ac-SDKP), proteomic technology, and cloning techniques. In aim II we will 1) perform a more extensive examination of the structural activity of Ac-SDKP in order to a) develop potent antagonists that lack partial agonistic activity and b) improve the affinity of the radio-iodinated peptide; and 2) characterize the Ac-SDKP receptor in fibroblasts and macrophages (rat and human), using [ l]-Hpp- Aca-SDKP and newly developed antagonists; and 3) compare rat cardiac fibroblasts and human cardiac fibroblasts for the inhibitory effect of Ac-SDKP or analogues on collagen synthesis and proliferation. In aim we will study whether Ac-SDKP receptor activity depends on mechanisms closely linked to the regulation of receptor internalization. In aim IV We will determine 1) the effect of Ac-SDKP on the non-receptor tyrosine kinase Src and HB-EGF on Ang II and ET-1-stimulated transactivation of the EGFR; 2) whether Ac-SDKP inhibits the effects of calcium ionophores or EGF on p42/44 MAPK and collagen synthesis; 3) whether PLC, EGFR, cSrc, calmodulin kinase or IP3 inhibitors attenuate MAPK activity and collagen synthesis to the same extent as Ac-SDKP in response to Ang II or ET-1; and 4) whether inhibition of MAP kinase activation by Ac- SDKP is mediated by MAP kinase phosphatase-1, using selective inhibitors of phosphatases and specific SiRNAs. This project will provide important new information on the d the mechanism of action of Ac-SDKP. Consequently, it will identify another component (Ac-SDKP) as part of the multiple mediators participating in the cardioprotective effects of ACEi in hypertension.

描述（由申请人提供）：高血压是心血管和肾脏疾病的主要危险因素。炎症和细胞外基质（EM）成分对高血压患者的动脉、心脏和肾脏等最终靶器官的生理和功能产生负面影响。阻断血管紧张素转换酶 (ACE) 会降低血管紧张素 II (Ang II) 并增加激肽，从而减少心血管炎症、肥大和胶原蛋白。 ACE 抑制剂 (ACEi) 会增加血浆 Ac-SDKP，Ac-SDKP 是血浆和组织中细胞增殖的负调节因子。在高血压和心力衰竭中，Ac-SDKP 可防止主动脉、肾脏和左心室 (LV) 中的单核细胞/巨噬细胞浸润和纤维化。凭借其抗纤维化和抗炎作用，Ac-SDKP能够改善高血压、糖尿病和其他肾脏疾病实验模型的肾功能。然而，Ac-SDKP 的心血管和肾脏作用所涉及的机制或受体尚未完全了解。我们假设 Ac-SDKP 通过位于质膜上的特定受体对高血压患者的心血管和肾脏系统发挥抗炎和抗纤维化作用，从而有助于保护终末器官。在目标 I 中，我们将使用药理学工具 [1]Hpp-Aca-SDKP、5(6)FAM-SDKP 和 Ac-SDKP 的新类似物、蛋白质组技术和克隆技术来鉴定和表征 Ac-SDKP 受体。在目标 II 中，我们将 1) 对 Ac-SDKP 的结构活性进行更广泛的检查，以便 a) 开发缺乏部分激动活性的有效拮抗剂，b) 提高​​放射性碘肽的亲和力； 2) 使用[1]-Hpp-Aca-SDKP和新开发的拮抗剂表征成纤维细胞和巨噬细胞（大鼠和人）中的Ac-SDKP受体； 3)比较大鼠心脏成纤维细胞和人心脏成纤维细胞Ac-SDKP或类似物对胶原合成和增殖的抑制作用。我们的目的是研究 Ac-SDKP 受体活性是否依赖于与受体内化调节密切相关的机制。在目标 IV 中，我们将确定 1) Ac-SDKP 对非受体酪氨酸激酶 Src 和 HB-EGF 对 Ang II 和 ET-1 刺激的 EGFR 反式激活的影响； 2) Ac-SDKP是否抑制钙离子载体或EGF对p42/44 MAPK和胶原蛋白合成的影响； 3) PLC、EGFR、cSrc、钙调蛋白激酶或IP3抑制剂是否会与Ac-SDKP响应Ang II或ET-1一样减弱MAPK活性和胶原蛋白合成； 4) 使用选择性磷酸酶抑制剂和特异性 SiRNA，研究 Ac-SDKP 对 MAP 激酶激活的抑制是否是由 MAP 激酶磷酸酶-1 介导的。该项目将提供有关 Ac-SDKP 作用机制的重要新信息。因此，它将确定另一种成分 (Ac-SDKP) 作为参与 ACEi 对高血压心脏保护作用的多种介质的一部分。

项目成果

Angiotensin II-induced dilated cardiomyopathy in Balb/c but not C57BL/6J mice.

• DOI: 10.1113/expphysiol.2011.057612
• 发表时间: 2011-08
• 期刊: Experimental physiology
• 影响因子: 2.7
• 作者: Peng H;Yang XP;Carretero OA;Nakagawa P;D'Ambrosio M;Leung P;Xu J;Peterson EL;González GE;Harding P;Rhaleb NE
• 通讯作者: Rhaleb NE