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: 7122069
• 负责人: CINTIA S. DE PAIVA
• 金额: $ 5.8万
• 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/7122069
• 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的紧密连接（TJ）对实验性干眼（EDE）的反应，并特别强调角膜上皮屏障功能的扰动。角膜上皮屏障将被评估为俄勒冈绿色葡聚糖的摄取，这是一种高分子量荧光分子（70KDA），它将与EDE前后的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响应EDE的TJ和LEP表达的作用。在预防角膜屏障功能障碍时，将评估应力相关途径/介体的特定抑制剂的有效性。在小鼠中，将通过药理学抑制撕裂分泌，并在具有较低湿度和持续的空气草稿的环境控制室中放置泪液分泌，并将其放置在小鼠中。实时PCR将用于评估促炎性介质的基因表达。 TJ和LEP表达将通过激光扫描共聚焦显微镜和免疫染色来评估。蛋白质印迹，ELISA和Luminex多重分析将评估炎性介质的表达。

项目成果

Stress-activated protein kinase signaling pathways in dry eye and ocular surface disease.

干眼和眼表疾病中的应激激活蛋白激酶信号通路。

• DOI: 10.1016/s1542-0124(12)70244-6
• 发表时间: 2005
• 期刊: The ocular surface
• 作者: Pflugfelder,StephenC;dePaiva,CintiaS;Tong,Louis;Luo,Lihui;Stern,MichaelE;Li,De-Quan
• 通讯作者: Li,De-Quan