细胞自噬在角膜缘干细胞稳态维持和功能障碍中的作用与机制

Limbal stem cells are ultimately responsible for the corneal epithelium under normal circumstances and during wound healing. Our preliminary studies showed that LSCs exhibited high autophagy levels and autophagy was involved in LSCD development. To investigate the function of autophagy in LSCs homeostasis and dysfunction, current project will be carried out to determine, 1) The effects of autophagy on LSCs in vitro; 2) The effects of autophagy in LSCs on the corneal homeostasis in vivo; 3) The relationship between autophagy and LSCD; 4) The feasibility of therapeutics targeting on autophagy for LSCD. This project will help illustrate the mechanism of the homostasis of LSCs, and to shed new light on the treatment of LSCD. It will also help improve the protocol of ex vivo expansion of corneal epithelial stem cells, as well as the corneal epithelium tissue engineering.

角膜缘干细胞（Limbal stem cell，LSC）是角膜上皮生发的源泉，也是角膜屏障功能维持的必要条件。我们的前期研究表明，LSCs中自噬水平较高，且抑制自噬与角膜缘干细胞功能障碍(limbal stem cell dysfunction，LSCD)发生相关。为了深入探究自噬影响LSCs的分子机理，本项目将进行如下研究(一)离体条件下自噬活性变化对LSCs表型的影响；（二）体内环境中LSCs自噬活性变化对角膜生理功能的影响；（三）LSCD发生过程中LSCs内自噬的时空动态变化特征；（四）自噬激动剂对LSCD治疗作用评价。本研究的开展有利于阐明LSCs稳态维持的机理，为LSCD的治疗提供新的思路和方法，也将有助于改善LSCs体外扩增技术以及组织工程角膜上皮构建技术。

睡眠缺乏是导致角膜缘干细胞功能障碍的重要因素之一，本项目为了探讨睡眠缺乏对角膜缘干细胞的影响及其作用机制，进行了如下研究（一）短期睡眠缺乏导致的角膜上皮细胞全基因组表达变化并进行聚类分析；（二）基因和蛋白分析确认了短期睡眠缺乏对角膜缘干细胞基因表达的影响；（三）短期睡眠缺乏对角膜缘干细胞氧化应激和抗氧化能力的调节作用；（四）外源H2O2对角膜缘干细胞增殖的影响；（五）抗氧化剂L-谷胱甘肽对角膜缘干细胞增殖的抑制作用；（六）明确PI3K/AKT信号通路在ROS介导的CEPCs过度增殖中起关键作用；（七）长期睡眠缺乏对角膜缘干细胞增殖能力的影响。本项目阐明了角膜缘干细胞功能障碍的分子机理，并为该种疾病的治疗提供了新的靶点。