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.

项目描述（由申请人提供）：本项目将研究紧密连接（TJ）对实验性干眼（EDE）在角膜上皮上的反应，特别强调角膜上皮屏障功能的扰动。角膜上皮屏障将通过俄勒冈绿葡聚糖（Oregon Green Dextran，一种高分子量荧光分子（70kda））的摄取来评估，并将在EDE治疗1、3、5、7、10和12天前后与TJ蛋白和晚期包膜蛋白（LEP）的表达相关。本研究还将评估促炎信号介质在EDE反应中对TJ和LEP表达的作用。应激相关通路/介质的特异性抑制剂在预防角膜屏障功能障碍中的有效性将被评估。用东莨菪碱对泪液分泌进行药理学抑制，并将其放置在低湿度和恒定气流的环境控制室中，从而诱导小鼠干眼症。Real time PCR将用于评估促炎介质的基因表达。TJ和LEP的表达将通过激光扫描共聚焦显微镜和免疫染色进行评估。促炎介质的表达将通过Western Blot、ELISA和Luminex Multiplex Assay进行评估。