PROTEIN KINASE C & CELL CYCLE IN VASCULAR INJURY

描述(改编自《调查者摘要》)：本申请 第一次获奖研究了蛋白激酶C的激活在 球囊导管损伤后即刻早期基因的表达 大鼠胸主动脉(BAL)。增强的平滑肌细胞(SMC) 增殖和迁移是血管内膜增厚的主要原因。 大鼠胸主动脉BAL检查结果。施加性伸展 在BAL中，SMC是不可避免的，这会产生更大的SMC 与单纯的内皮剥脱相比，增殖反应。 蛋白激酶C(PKC)抑制剂和钙通道阻滞剂是已知的 在体外调节SMC增殖，后一种化合物减少 BAL后在体内的增殖。这样做的根本目的是 应用于描绘早期钙依赖信号转导 牵张诱导静止SMC激活的机制 在体内，这与体液因素有关，导致 激活SMC细胞周期。前两个目标以BAL为中心 牵张对原位灌流的主动脉PKC的调节作用 准备工作。目标1包括时间进程和钙依赖 牵张诱导的激活和膜移位 钙敏感和不敏感的PKC亚型。目标2描述了 两相产生PKC辅因子的时间进程和要求 甘油二酯和肌醇3，4，5-三磷酸。这些发现将 在目标3中使用SMC在体外进行扩展。该目标将检查 G_1期特定蛋白激酶C的转位和下调 在体外的细胞周期中。血管壁中的SMC具有 生长受类肝素抑制的“收缩”表型 与细胞外基质相关的分子。与.相比 体外扩增合成的SMC，灌流模型更多 与拉伸相关的原位信号转导机制相关 更有可能模仿静止的最早的激活反应 活体内的SMC。PKC和细胞周期的研究是公认的在IN 体外系统反映了持续存在的细胞生长控制 增殖性，其中拉伸的建模可能受到限制。 然而，这些研究是随后调查的基础 蛋白激酶C在分化过程中细胞周期调控中的作用 血管壁SMC。

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.