Central GRK5 modulation of Angiotensin II receptor expression in heart failure

GRK5 对心力衰竭中血管紧张素 II 受体表达的中枢调节

• 批准号: 8531707
• 负责人: Karla Haack
• 金额: $ 5.39万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8531707
• 关键词: Address; Anesthesia procedures; Angiotensin II; Angiotensin II Receptor; Animal Model; Animals; Attenuated; Baroreflex; Brain; Brain Stem; Brain region; Cardiac; Cause of Death; Cell Line; Cells; Chronic; Complex; Coronary artery; Data; Diagnosis; Disease; Dissociation; Dominant-Negative Mutation; Echocardiography; Excretory function; Exercise; Frequencies; G protein coupled receptor kinase; G-Protein-Coupled Receptors; GRK; GRK5 gene; Genetic Transcription; Heart Rate; Heart failure; Hypertension; Hypothalamic structure; In Vitro; Injection of therapeutic agent; Laboratories; Lead; Ligation; Link; Measures; Mediating; Mus; Muscle; Nerve; Neurons; Norepinephrine; Nuclear; Patients; Phosphorylation; Phosphotransferases; Plasma; Plasmids; Play; Process; Quality of life; Rattus; Receptor, Angiotensin, Type 1; Recycling; Regulation; Role; Small Interfering RNA; Subfamily lentivirinae; Techniques; Testing; Therapeutic; Training; Transcriptional Regulation; Ubiquitin-Proteasomal Pathway; United States; Up-Regulation; effective therapy; hemodynamics; improved; indexing; overexpression; paraventricular nucleus; pressure; prevent; protein expression; receptor; receptor expression; receptor upregulation; research study; response; sedentary; therapeutic development; therapeutic target; urinary

DESCRIPTION (provided by applicant): This proposal will address the following specific aims: (1) Determine the role(s) of central GRK5 in the regulation of AT1R (Angiotensin II type 1 Receptor) expression under normal conditions and during Chronic Heart Failure (CHF). (2) Identify the mechanism(s) of modulation of central AT1R and GRK5 following Exercise Training (ExT) in CHF animals. (3) Determine the role of cytosolic and nuclear GRK5 in the transcriptional regulation of AT1R by I?B¿ and NF-?B. In Specific Aim 1 we will induce CHF by coronary artery ligation. Overexpression of GRK5 will be targeted to the RVLM or PVN by lentiviral injection. To determine the effects of GRK5 knockdown on AT1R expression and changes in sympatho-excitation, we will utilize both commercially available GRK5 KO mice and lentiviral packaged siRNA against GRK5 that will be injected into the RVLM or PVN. Urinary excretion of norepinephrine (NE), plasma NE and Ang II will be measured in all animal groups. Arterial pressure and heart rate will be continuously recorded in order to derive additional indices of sympatho-excitation and to determine arterial baroreflex function. Sympathetic nerve activity will be directly recorded under anesthesia in terminal experiments. In all animals cardiac function will be evaluated serially by high-frequency echocardiography. In Specific Aim 2, we will induce CHF in GRK5KO mice that are either sedentary or ExT as well as utilize CHF rats in which GRK5 has been silenced in the PVN or RVLM using siRNA lentivirus. Following ExT, sympathetic and baroreflex function will be evaluated in a similar fashion as in Specific Aim 1. We will test Specific Aim 3 in CATH.a neurons, utilizing both overexpression and silencing techniques with a GRK5 plasmid and siRNA to examine the subcellular localization of AT1R, I?B¿, NF-?B, and GRK5 following Ang II stimulation. We will also perform parallel studies using a K215R dominant negative GRK5 construct to determine if the GRK5/AT1R/I?B¿/NF-?B interaction is kinase-dependent.

描述（由适用提供）：该提案将解决以下具体目的：（1）在正常条件和慢性心力衰竭（CHF）期间确定中央GRK5在调节AT1R（血管紧张素II型1型受体）表达中的作用。 （2）确定CHF动物运动训练后中央AT1R和GRK5调制的机制。 （3）确定胞质和核GRK5在i？b¿和nf- b的AT1R转录调控中的作用。在特定目标1中，我们将通过冠状动脉结扎诱导瑞士法郎。 GRK5的过表达将通过慢病毒注射针对RVLM或PVN。为了确定GRK5敲低对AT1R表达的影响以及交感神经激发的变化，我们将利用将注射到RVLM或PVN中的市售GRK5 KO小鼠和慢病毒包装的siRNA对GRK5的影响。尿极端。去甲肾上腺素（NE），血浆NE和ANG II将在所有动物群中进行测量。动脉压和心率将连续记录，以得出其他交感神经引起的指标并确定动脉压力反射功能。在终末实验中，将在麻醉下直接记录交感神经活动。在所有动物心脏 功能将通过高频超声心动图串行评估。在特定的目标2中，我们将在久坐的或ext的GRK5KO小鼠中诱导CHF，并利用使用siRNA慢病毒在PVN或RVLM中沉默的CHF大鼠。在Ext之后，将以与特定目标1相似的方式评估交感神经和Baroreflex功能。我们将使用GRK5质粒和siRNA使用过表达和沉默技术来测试特定的目标3，以检查AT1R的亚细胞定位，并检查AT1R的亚细胞位置，i？b？，nf-？b？，nf-？b ang ii simulation simulation simulation simulation simulation simulation。我们还将使用K215R显性的负GRK5构建体进行并行研究，以确定GRK5/AT1R/I？b¿/nf- b的相互作用是否依赖激酶。