ARTERIAL INJURY AND SMOOTH MUSCLE CELL SIGNALING

描述(改编自《调查员摘要》)：本提案将 确定重要的细胞内信号通路 动脉损伤后SMC的复制。第一组实验 将记录MAP激酶通路的激活，即p42/44erk 气球损伤的大鼠颈动脉。激活这一途径将 局部应用MEK1抑制剂在体内被抑制及其作用 对SMC复制进行了量化。此外，激活的效果 上游激活剂转染细胞对SMC生长的影响 并测量它们的复制能力以响应已知的 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SMC复制的影响 大鼠颈动脉。最初，mkp1/2表达式将记录在 受伤后的不同时间。过钒酸盐可抑制MKP-1的表达 它的表达被反义寡核苷酸阻断。复制 颈动脉SMC对已知激动剂的作用将被量化。第三 AIM将确定MAP激酶p42/44erk是否被FGF2和 PDGF。大鼠动脉将以一种已知的方式剥离内皮 诱导最小的SMC复制，然后FGF2和PDGF将 管理。P42/44erk活性和MKP-1的表达 以这些动脉的SMC进行测量。最后一组实验将 记录细胞周期蛋白D和E活性的时间进程以及 颈动脉损伤后细胞周期蛋白抑制因子p21和p27的表达。进一步 研究将通过以下方法评估p27对体内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。在其他 研究抑制因子p27在SMC中的过度表达及其在血管内皮生长中的作用 抑制SMC复制，当受到有丝分裂刺激时， 量化的。这些研究将提供有关信号的重要数据。 被认为在SMC中活跃的通路，并确定它们的激活是否 对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.