PROTEIN KINASE C & CELL CYCLE IN VASCULAR INJURY

蛋白激酶C

• 批准号: 6043840
• 负责人: CAROLYN J SMITH
• 金额: $ 11.21万
• 依托单位: NEW YORK MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2001-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6043840
• 关键词: DNA replication; aorta; biological signal transduction; blood vessel disorder; calcium channel; calcium channel blockers; cell cycle; cell growth regulation; diacylglycerols; enzyme activity; gel electrophoresis; immunoprecipitation; injury; inositol phosphates; isozymes; laboratory rat; northern blottings; protein kinase C; regulatory gene; tissue /cell culture; vascular smooth muscle; western blottings

DESCRIPTION (Adapted from Investigator's Abstract): This application for a FIRST Award examines the role of activation of protein kinase C in immediate early gene expression following balloon catheter-induced injury of the rat thoracic aorta (BAL). Enhanced smooth muscle cell (SMC) proliferation and migration account for the intimal thickening which results from BAL of the rat thoracic aorta. The imposition of stretch upon SMC is inevitable in BAL, which produces a greater SMC proliferative response as compared to endothelial denudation alone. Protein kinase C (PKC) inhibitors and calcium channel blockers are known to modulate SMC proliferation in vitro, and the latter compounds reduce proliferation following BAL in vivo. The fundamental goal of this application is to delineate early calcium dependent signal transduction mechanisms responsible for stretch-induced activation of quiescent SMC in vivo, which in association with humoral factors, results in activation of the SMC cell cycle. The first two Aims center upon BAL stretch-induced regulation of PKC in an in situ perfused aortic preparation. Aim 1 covers the time course and calcium dependence for stretch-induced activation and membrane translocation of both the calcium sensitive and insensitive PKC isoforms. Aim 2 characterizes the time course and requirements for biphasic generation of the PKC cofactor diacylglycerol and inositol 3,4,5-trisphosphate. These findings will be extended using SMC in vitro in Aim 3. This Aim will examine translocation and down regulation of specific PKCs during the G1 phase of the cell cycle in vitro. The SMC in the vessel wall possesses a "contractile" phenotype whose growth is inhibited by heparinoid molecules associated with the extracellular matrix. Compared to proliferating "synthetic" SMC in vitro, the perfusion model is more relevant in that stretch-related signal transduction mechanisms in situ are more likely to mimic the earliest activation response of quiescent SMC in vivo. It is recognized the PKC and cell cycle studies in the in vitro system reflects growth control in cells that are persistently proliferative and in which the modeling of stretch may be limited. However, these studies serve as a basis of subsequent investigation of the role of PKC on the regulation of the cell cycle in differentiated vessel wall SMC.

描述（改编自研究者摘要）：本申请 FIRST 奖研究了蛋白激酶 C 激活在 球囊导管诱导损伤后立即早期基因表达 大鼠胸主动脉（BAL）。 增强型平滑肌细胞（SMC） 增殖和迁移导致内膜增厚， 大鼠胸主动脉 BAL 的结果。 施加拉伸 BAL 中 SMC 是不可避免的，它会产生更大的 SMC 与单独的内皮剥脱相比，增殖反应。 蛋白激酶 C (PKC) 抑制剂和钙通道阻滞剂是已知的 体外调节 SMC 增殖，后者化合物可减少 BAL 体内增殖。 本次活动的根本目标是 应用是描绘早期钙依赖性信号转导 拉伸诱导静态 SMC 激活的机制 在体内，与体液因素相关，导致 SMC 细胞周期的激活。 前两个目标以 BAL 为中心 原位灌注主动脉中牵拉诱导的 PKC 调节 准备。 目标 1 涵盖了时间进程和钙依赖性 拉伸诱导的激活和膜易位 钙敏感和不敏感 PKC 亚型。 目标 2 的特点是 PKC 辅因子双相生成的时间过程和要求 二酰基甘油和肌醇3,4,5-三磷酸。 这些发现将 在目标 3 中使用体外 SMC 进行扩展。该目标将检查 G1 期特定 PKC 的易位和下调 体外细胞周期。 血管壁中的 SMC 具有 “收缩”表型，其生长被类肝素抑制 与细胞外基质相关的分子。 相比 体外增殖“合成”SMC，灌注模型更 与拉伸相关的原位信号转导机制相关 更有可能模仿静态的最早激活反应 体内的SMC。 PKC和细胞周期研究在国际上得到认可 体外系统反映了持续存在的细胞的生长控制 增殖性的，其中拉伸的建模可能受到限制。 但这些研究可以作为后续调查的基础 PKC对分化细胞周期的调节作用 血管壁 SMC。