CELL SIGNAL TRANSDUCTION IN CORNEAL WOUND HEALING

角膜伤口愈合中的细胞信号转导

• 批准号: 2710926
• 负责人: Haydee E.P. Bazan
• 金额: $ 20.43万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-30 至 2002-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2710926
• 关键词: DNA replication; biological signal transduction; cell growth regulation; cell proliferation; corneal epithelium; enzyme activity; gene expression; genetic transcription; growth factor receptors; high performance liquid chromatography; imaging /visualization /scanning; immunoprecipitation; laboratory rabbit; mitogen activated protein kinase; northern blottings; organ culture; phosphatidylinositol 3 kinase; phospholipids; phosphorylation; protein isoforms; protein kinase C; western blottings; wound healing

DESCRIPTION: (adapted from the applicant s abstract) Corneal re-epithelization after trauma or surgery is fundamental to restore normal vision. The PI plans to test the hypothesis that phosphotidylinositol-3-kinase (PI-3K) and protein kinase C (PKC) are essential steps in the signal transduction pathway leading to repair. These pathways transmit stimuli from the cell surface to the nuclei in a cascade of events that involve mitogen-activated protein kinase (MAPK). The debridement of the corneal epithelial layer will be used as a model of wound healing. The PI proposes to test if PKC isoforms can directly activate transcription by translocating to the nuclei or indirectly by activating the MAPK cascade, regulating cell proliferation and wound healing. She also proposes to perform studies to establish how the receptors for epidermal growth factor (EGF) and keratinocyte growth factor (KGF) associate with PI-3K and to define which PKC isoforms are targets of PI-3K. Selective inhibitors will be used to correlate the studies with wound closure. Analytical procedures such as HPLC and FPLC will be used. Identification of pathway components will be accomplished by Western blot with specific antibodies, and kinase assays will be performed in enzyme immunocomplexes and quantified by phosphor imaging. The results obtained will define the involvement of PI-3K, PKC and MAPK in corneal wound healing and help target drugs to selective steps of signal transduction to modulate corneal epithelial repair.

描述：（改编自申请人的摘要）角膜 创伤或手术后的上皮再生是恢复正常的基础 想象。 PI 计划检验以下假设： 磷脂酰肌醇-3-激酶 (PI-3K) 和蛋白激酶 C (PKC) 是 导致修复的信号转导途径中的重要步骤。 这些 通路以级联方式将刺激从细胞表面传递到细胞核 涉及丝裂原激活蛋白激酶 (MAPK) 的事件。 这 角膜上皮层清创术将用作伤口模型 康复。 PI建议测试PKC亚型是否可以直接激活 通过易位到细胞核或间接通过激活 MAPK 级联，调节细胞增殖和伤口愈合。 她还 提议进行研究以确定表皮受体如何 生长因子（EGF）和角质形成细胞生长因子（KGF）与 PI-3K 并定义哪些 PKC 同工型是 PI-3K 的目标。 选择性 抑制剂将用于将研究与伤口闭合相关联。 将使用 HPLC 和 FPLC 等分析程序。 鉴定 途径组分将通过具有特定的蛋白质印迹来完成 抗体和激酶测定将在酶免疫复合物中进行 并通过磷光体成像进行量化。 获得的结果将定义 PI-3K、PKC 和 MAPK 参与角膜伤口愈合并帮助靶向 药物通过信号转导的选择性步骤来调节角膜 上皮修复。