Vascular tone & cAMP phosphodiesterase in angioplasty

血管张力

• 批准号: 6922843
• 负责人: JOHN QUILLEY
• 金额: $ 31.3万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6922843
• 关键词: 3' 5' cyclic nucleotide phosphodiesterase; aorta; biological signal transduction; cardiovascular injury; cell proliferation; cyclic AMP; enzyme activity; enzyme induction /repression; immunocytochemistry; in situ hybridization; intraluminal angioplasty; isozymes; laboratory rat; phosphodiesterase inhibitors; protein kinase A; vascular endothelium; vascular smooth muscle; vasodilators; vasomotion; vasospasm

EXCEED THE SPACE PROVIDED. Enhanced smooth muscle cell (SMC) proliferation and migration account for intitnal thickeningwhich follows balloon catheter de-endothelialization(BAL) of rat aorta. Recent studies show activation of cAMP-dependent protein kinase (PKA) by specific inhibition of cAMP phosphodiesterases PDE3 and/or PDE4 reduces SMC proliferation/ migration and lesions after angioplasty. In addition to changes in SMC growth, contractility may be affected by injury. Overall hypothesis: BAL leads to enhanced expression and activity of specific cAMP PDEs in both SMC and vessel wall-associated inflammatory cells, which then affects SMC cyclic nucleotide levels, protein phosphorylation and contractility. The project will first identify which high affinity, cAMP-selective PDE3 and PDE4 isoforms are upregul- ated in the BAL-injured rat aorta or growth factor-stimulated rat aortic SMC, while later aims assess the impact of PDE upregulationon cAMP-dependent phosphorylation,vessel contractility, and cellular locale. Aim 1 A: Determine the time course of protein expression for PDE3A/3B and PDE4A/4B/4D genes followingBAL in vivo. Pilot data for aortic medial SMC show that BAL is associated with biphasic increases in PDE4BimRNA, and smaller (PDE3B) or no changes in other genes (PDE3A, 4D). Aim IB: To determine the time course of PDE4B, PDE4D and PDE3A protein splice variants in RASMC stimulatedwith serum, PDGF-BB or bFGF. Aim 2A: Determine the time course for PDE inhibitor enhancementof PKA activity, or PKA-dependent phosphorylation of a vasodilator-sensitive substrate VASP in SMC. These indicesof intracellular cAMP will be used to determine if inhibition of overexpressed PDE3/4 in BAL restores or enhances beta-agonist and forskolin-dependent activation. Aim 2B: Determine the impact of PDE3/4 upregulation in vitro on VASP phosphorylation and PKA activity. Aim 3: Characterize contractility of the BAL aorta at 24 hr and 1-2 weeks after injury with various vasodilators plus/minus PDE inhibitors. Aim 4: Identify at 24 hr and 7- 14 days after BAL the aortic cellular specificityof PDE expression by immunohistochemistry and in situ hybridizati- on. The increase in PDE3/4 is predicted to reduce vasorelaxation produced by agents which increase cAMP and cGMP levels. PDE overexpression favors vasospasm, which may affect vessel wall remodeling.Upregulation of specific PDEs represents an importantresponse to injury that may serve as a therapeutictarget in restenosis and hypertension. PERFORMANCE SITE ========================================Section End===========================================

超出所提供的空间。大鼠主动脉球囊导管去内皮化（BAL）后平滑肌细胞（SMC）增殖和迁移增强导致内膜增厚。最近的研究表明，通过特异性抑制 cAMP 磷酸二酯酶 PDE3 和/或 PDE4 来激活 cAMP 依赖性蛋白激酶 (PKA)，可减少血管成形术后的 SMC 增殖/迁移和病变。除了 SMC 生长的变化之外，收缩性也可能受到损伤的影响。总体假设：BAL 导致 SMC 和血管壁相关炎症细胞中特定 cAMP PDE 的表达和活性增强，从而影响 SMC 环核苷酸水平、蛋白质磷酸化和收缩性。该项目将首先确定哪些高亲和力、cAMP 选择性 PDE3 和 PDE4 亚型在 BAL 损伤的大鼠主动脉或生长因子刺激的大鼠主动脉 SMC 中上调，随后的目标是评估 PDE 上调对 cAMP 依赖性磷酸化、血管收缩性和细胞部位的影响。目标 1 A：确定体内 BAL 后 PDE3A/3B 和 PDE4A/4B/4D 基因蛋白质表达的时间过程。主动脉内侧 SMC 的试验数据显示，BAL 与 PDE4BimRNA 的双相增加有关，而其他基因（PDE3A、4D）的变化较小（PDE3B）或没有变化。目标IB：确定用血清、PDGF-BB或bFGF刺激的RASMC中PDE4B、PDE4D和PDE3A蛋白剪接变体的时程。目标 2A：确定 PDE 抑制剂增强 PKA 活性或 SMC 中血管舒张剂敏感底物 VASP 的 PKA 依赖性磷酸化的时间过程。这些细胞内 cAMP 指数将用于确定 BAL 中过度表达的 PDE3/4 的抑制是否恢复或增强 β-激动剂和毛喉素依赖性激活。目标 2B：确定体外 PDE3/4 上调对 VASP 磷酸化和 PKA 活性的影响。目标 3：使用各种血管扩张剂加上/减去 PDE 抑制剂，表征损伤后 24 小时和 1-2 周时 BAL 主动脉的收缩性。目标 4：通过免疫组织化学和原位杂交鉴定 BAL 后 24 小时和 7-14 天 PDE 表达的主动脉细胞特异性。 PDE3/4 的增加预计会减少由增加 cAMP 和 cGMP 水平的药物产生的血管舒张作用。 PDE 过度表达有利于血管痉挛，可能影响血管壁重塑。特定 PDE 的上调是对损伤的重要反应，可作为再狭窄和高血压的治疗靶点。 表演网站==========================================章节结束===============================================

项目成果

Increased phosphodiesterase 3A/4B expression after angioplasty and the effect on VASP phosphorylation.

• DOI: 10.1016/j.ejphar.2008.05.016
• 发表时间: 2008-08
• 期刊: European journal of pharmacology
• 影响因子: 5
• 作者: Hong Zhao;Qizhi Guan;Carolyn J. Smith;J. Quilley

Differential effects of phosphodiesterase PDE-3/PDE-4-specific inhibitors on vasoconstriction and cAMP-dependent vasorelaxation following balloon angioplasty.

磷酸二酯酶 PDE-3/PDE-4 特异性抑制剂对球囊血管成形术后血管收缩和 cAMP 依赖性血管舒张的不同影响。

• DOI: 10.1152/ajpheart.00419.2006
• 发表时间: 2007
• 期刊: American journal of physiology. Heart and circulatory physiology
• 作者: Zhao,Hong;Quilley,John;Montrose,DavidC;Rajagopalan,Swarna;Guan,Qizhi;Smith,CarolynJ
• 通讯作者: Smith,CarolynJ

Effect of tempol on renal cyclooxygenase expression and activity in experimental diabetes in the rat.

tempol 对实验性糖尿病大鼠肾环氧合酶表达和活性的影响。

• DOI: 10.1124/jpet.104.076927
• 发表时间: 2005
• 期刊: The Journal of pharmacology and experimental therapeutics.
• 作者: Li,Jing;Chen,Yu-Jung;Quilley,John
• 通讯作者: Quilley,John