ARTERIAL INJURY AND SMOOTH MUSCLE CELL SIGNALING

描述（改编自研究者摘要）：本提案将 确定细胞内信号通路，这是重要的， 动脉损伤后SMC复制。 第一组实验 将记录MAP激酶通路的激活，即p42/44 erk在 膨胀的受损大鼠颈动脉。 激活这一途径将 在体内被局部施用的MEK 1抑制剂抑制， 对SMC复制的影响。 此外，激活的效果 p46 sapk通过上游激活剂抑制细胞对SMC生长的影响 的p46 sapk和测量他们的复制能力，以应对已知的 有丝分裂原 第二个具体目标将决定 丝裂原活化蛋白激酶磷酸酶（MKP-1）对平滑肌细胞复制的影响 大鼠颈动脉 最初，MKP 1/2表达将记录在 受伤后的几次。 MKP-1将被过钒酸盐抑制， 用反义寡核苷酸阻断其表达。 复制 然后将定量颈动脉SMC对已知激动剂的反应。 第三 目的是确定MAP激酶，p42/44 erk是否被FGF 2激活， PDGF。 大鼠动脉将以已知的方式剥脱内皮， 诱导最低限度的SMC复制，然后将FGF 2和PDGF 管理。 检测p42/44 erk的活性和MKP-1的表达， 在这些动脉的SMC中测量。 最后一组实验将 记录细胞周期蛋白D和E活性的时间过程以及 细胞周期蛋白抑制剂，p21和p27在颈动脉损伤后的时间。 进一步 研究将评估p27在体内对SMC复制的影响， 用反义寡核苷酸抑制其表达，然后 测量内膜SMC对已知有丝分裂原的反应能力。 换句 研究了抑制剂p27在SMC中的过度表达及其在 抑制SMC复制，当用促有丝分裂刺激物激发时， 定量。 这些研究将提供有关信号传导的重要数据。 被认为在SMC中活跃的途径，并确定它们的激活是否是 对于SMC复制至关重要。

DESCRIPTION (Adapted from Investigator's Abstract): This proposal will determine the intracellular signaling pathways that are important for the replication of SMC after arterial injury. The first group of experiments will document the activation of the MAP kinase pathways, namely p42/44erk in the ballooned injured rat carotid arteries. Activation of this pathway will be inhibited in vivo with a topically applied MEK1 inhibitor and the effect on SMC replication quantitated. Furthermore, the effect of activation of the p46sapk on SMC growth by transfecting cells with the upstream activator of p46sapk and measuring their ability to replicate in response to known mitogens. The second specific aim will determine the role of mitogen-activated protein kinase phosphatases (MKP-1) on SMC replication in rat carotid arteries. Initially MKP1/2 expression will be documented at various times after injury. MKP-1 will be inhibited with pervanadate and its expression blocked with an antisense oligonucleotide. The replication of carotid artery SMC to known agonists will then be quantitated. The third aim will determine if the MAP kinases, p42/44erk are activated by FGF2 and PDGF. Rat arteries will be denuded of endothelium in a manner known to induce a minimal SMC replication, and then FGF2 and PDGF will be administered. The activity of p42/44erk and the expression of MKP-1 will be measured in the SMC of these arteries. The final group of experiments will document the time course of cyclin D and E activity and the presence of cyclin inhibitors, p21 and p27 at times after carotid injury. Further studies will evaluate the effect of p27 on SMC replication in vivo by inhibiting its expression with an antisense oligonucleotide and then measuring the ability of intimal SMC to respond to known mitogens. In other studies the inhibitor, p27 will be over expressed in SMC and its role in suppressing SMC replication, when challenged with a mitogenic stimulus, quantitated. These studies will provide important data on the signaling pathways thought to be active in SMC and determine if their activation is critical for SMC replication.