The stress of dry eye on corneal barrier function

干眼症对角膜屏障功能的压力

• 批准号: 6999263
• 负责人: CINTIA S. DE PAIVA
• 金额: $ 5.75万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6999263
• 关键词: cell membrane; confocal scanning microscopy; corneal epithelium; enzyme linked immunosorbent assay; gene expression; immunocytochemistry; immunologic assay /test; keratoconjunctivitis sicca; laboratory mouse; membrane activity; physiologic stressor; physiology; polymerase chain reaction; postdoctoral investigator; tight junctions; western blottings

DESCRIPTION (provided by applicant): The project will study the response of the tight junctions (TJ) to experimental dry eye (EDE) on the corneal epithelium with specific emphasis on the perturbation of corneal epithelial barrier function. Corneal epithelial barrier will be evaluated as the uptake of Oregon Green Dextran, a high molecular weight fluorescent molecule (70kda) and it will be correlated to the expression of TJ proteins and late envelope proteins (LEP) prior to and after 1,3,5,7,10 and 12 days of EDE. This study will also evaluate the role of pro-inflammatory signaling mediators on expression of TJ and LEP in response to EDE. Effectiveness of specific inhibitors of stress associated pathways/mediators will be evaluated in the prevention of corneal barrier dysfunction. The dry eye will be induced in mice by pharmacologic inhibition of tear secretion with scopolamine and placement in an environmentally controlled chamber with low humidity and a constant air draft. Real time PCR will be used to evaluate the gene expression of pro inflammatory mediators. TJ and LEP expression will be evaluated by laser scanning confocal microscopy and immunostaining. Pro inflammatory mediator expression will be evaluated by Western Blot, ELISA and the Luminex Multiplex Assay.

描述（由申请人提供）：该项目将研究角膜上皮上紧密连接（TJ）对实验性干眼（EDE）的反应，特别强调角膜上皮屏障功能的扰动。角膜上皮屏障将根据俄勒冈绿葡聚糖（一种高分子量荧光分子（70kda））的摄取进行评估，并将与 EDE 1、3、5、7、10 和 12 天之前和之后 TJ 蛋白和晚期包膜蛋白 (LEP) 的表达相关。这项研究还将评估促炎信号传导介质对 EDE 反应中 TJ 和 LEP 表达的作用。将评估应激相关途径/介质的特异性抑制剂在预防角膜屏障功能障碍方面的有效性。通过用东莨菪碱药理学抑制泪液分泌并将其放置在低湿度和恒定通风的环境控制室中，诱导小鼠干眼。实时 PCR 将用于评估促炎介质的基因表达。 TJ 和 LEP 表达将通过激光扫描共聚焦显微镜和免疫染色进行评估。将通过蛋白质印迹、ELISA 和 Luminex 多重测定来评估促炎介质表达。