CORNEAL ENDOTHELIAL REPAIR--ROLE OF LIPID MEDIATORS

本项目的总体目标是阐明 控制角膜内皮的伤口修复。 中央 在拟议的研究中探索的假设是，花生四烯酸 衍生的脂质介质合成响应损伤或 细胞外信号调节角膜的进展 内皮细胞通过其有丝分裂和分化周期。的 拟议研究的目的是： 1)鉴定角膜内皮的调节性脂质介质， 2)表征这种介质的生产有丝分裂和 分化内皮细胞， 3)确定受体依赖性信号转导是否受到调节， 有丝分裂和分化的内皮细胞， 4)探索受体后信号信号效应器通路， 脂质介质对角膜细胞有丝分裂和分化的调控 内皮细胞和 5)比较脂质介导的有丝分裂和分化的调节， 兔人角膜内皮细胞 这些问题将使用有丝分裂，分化和 组织培养兔角膜内皮细胞伤口愈合模型的建立 培养和使用人角膜内皮的器官培养模型。 调节有丝分裂和分化的脂质介质将是 使用细胞周期的细胞化学检测进行鉴定， 分化标志物 介质的生物合成和调节 受体介导的事件将使用生物化学示踪剂来确定， 生物合成酶免疫测定和第二类的生化测定 使者细胞内效应通路的参与将是 研究使用药理学探针改变信号转导 途径。 脂质介质改变兔的有丝分裂和分化 还将使用以下器官培养模型测试角膜内皮： 和营养不良的人角膜内皮。 获得的结果 这些研究将提出增强角膜内皮细胞的治疗策略， 意外或手术创伤后的内皮伤口修复。

The overall objective of this project is to elucidate the mechanisms governing wound repair in the corneal endothelium. The central hypothesis to be explored in the proposed studies, is that arachidonate derived lipid mediators synthesized in response to injury or to extracellular signals modulate the progression of the corneal endothelial cell through its cycle of mitosis and differentiation. The aims of the proposed study are to: 1) identify the modulatory lipid mediators of the corneal endothelium, 2) characterize the production of such mediators by mitotic and differentiating endothelial cells, 3) determine if receptor-dependent signal transduction is regulated in mitotic and differentiated endothelial cells, 4) explore post-receptor signal signal effector pathways that govern lipid mediator regulation of mitosis and differentiation in corneal endothelial cells and 5) compare lipid mediated regulation of mitosis and differentiation in rabbit human corneal endothelial cells. These questions will be investigated using mitotic, differentiated and wound closure models of rabbit corneal endothelial cells grown in tissue culture, and using organ culture models of human corneal endothelium. Lipid mediators regulating mitosis and differentiation will be identified using cyctochemical detection of cell cycle and differentiation markers. Biosynthesis of mediators and regulation of receptor mediated events will be determined using biochemical tracers, immunoassay of biosynthetic enzymes and biochemical assay of second messengers. The involvement of intracellular effector pathways will be investigated using pharmacological probes to alter signal transduction pathways. Lipid mediators found to alter mitosis and differentiation of rabbit corneal endothelium will also be tested using organ culture models of and dystrophic human corneal endothelium. The results obtained from these studies will suggest therapeutic strategies for enhancing corneal endothelial wound repair subsequent to accidental or surgical trauma.