力刺激调节LASIK术后圆锥角膜病变过程中MMP-2/9表达的分子机制研究

项目名称： 力刺激调节LASIK术后圆锥角膜病变过程中MMP-2/9表达的分子机制研究

项目编号： No.31300770

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 生物科学

项目作者： 宋婕

作者单位： 太原理工大学

项目金额： 23万元

中文摘要： LASIK术后角膜受力发生变化，MMP表达异常，胶原降解改变，角膜力学性能降低进而诱发圆锥角膜。机械拉伸能够诱导角膜细胞中MMP-2/9的表达，但其分子机制还不清楚。前期研究推测力刺激通过IL-1和整合素途径及下游的ERK-MAPK信号通路调控角膜细胞MMP-2/9的表达。本项目拟采用Flexcell 4000拉伸系统对体外培养的角膜细胞进行力刺激，用MAPK通路抑制剂、siRNA-MAPK、IL-1受体抑制剂、整合素抑制剂对角膜细胞相关信号途径进行干扰，结合力学加载，分别采用定量PCR、Western blot、明胶酶谱法检测处理前后角膜细胞中MMP-2/9相关基因表达及蛋白活性的变化，分析整合素、IL-1信号途径以及MAPK信号通路对力学加载诱导MMP-2/9基因表达的影响，初步阐明力刺激诱导角膜细胞MMP-2/9表达的分子机制，为研究圆锥角膜等LASIK术后并发症的发生机理提供参考。

中文关键词： 角膜细胞；机械拉伸；MMP-2；信号通路；

英文摘要： The decreased biomechanical properties as well as the abnormal MMP expression and collagen degradation after LASIK would induce postoperative complications including keratoconus. In our previous study, mechanical stretch was introduced to mimick the stress environment after LASIK, which resulted in an increase of MMP-2/9 expression in corneal cells in vitro. However, its molecular mechanism is still unclear. IL-1 and integrin signal pathway as well as the downstream ERK-MAPK pathway were suggested to participate in the mechanical stretch-induced MMP-2/9 expression in corneal cells. In light of this, Flexcell 4000 system was adopted to apply mechanical stretch on corneal cells in vitro in this study. Inhibitors of MAPK pathway, siRNA-MAPK, IL-1 receptor inhibitors as well as integrin inhibitors were incubated with corneal cells respectively to block the corresponding signal pathway. Mechanical stretch was then applied to the treated corneal cells. To analyze the effects of these pathways on the mechanical stretch-regulated MMP-2/9 activity, gene expression, protein content and activity of MMP-2/9 and its related proteins in the pathways were detected using real-time PCR, western blot and gelatin zymography. Our results can serve as the basis for better understanding the mechanism of keratoconus progression after

英文关键词： Keratocytes；Mechanical stretch；MMP-2；Signaling pathway；