Cui Wei, Bryant Michael R, Sweet Paula M, McDonnell Peter J
Doheny Eye Institute and the Department of Ophthalmology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.
Exp Eye Res. 2004 Feb;78(2):275-84. doi: 10.1016/j.exer.2003.10.007.
Scleral fibroblasts are involved in scleral remodeling during axial elongation in myopia. Mechanical load is a potent stimulator of gene expression. This study seeks to identify changes in gene expression of scleral fibroblasts in response to mechanical load and speculate on possible mechanisms of scleral remodeling in the development of myopia. Human scleral fibroblasts (HSFs) were mechanically stretched for 30 min and 24 hr. A gene microarray analysis was used to measure changes in gene expression. A total of 237 genes revealed differential and significant changes in expression (P<0.01) after 30 min of stretching. Of these, 28 unexpressed genes began to be expressed (turned on), while 31 expressed genes were no longer expressed (turned off). After 24 hr, 308 genes showed reproducible changes in expression (P<0.01), while 29 genes were turned on and 17 genes were turned off. After 30 min, 25 genes showed at least a threefold change in expression. These included genes for cell receptors, protein kinases, cell growth/differentiation factors, extracellular matrix (ECM) proteins, lipid metabolism, protein metabolism, transcription factors, binding proteins and water channels. After 24 hr, 21 genes showed at least a threefold change in expression. These included genes for cell receptors, protein kinases, cell growth/differentiation factors, lipid metabolism, ECM proteins, transcription factors, and carbohydrate metabolism. RT-PCR and Southern blotting confirmed the changes in expression of selected genes. In this study we identified a large number of early and late mechanical response genes in HSFs. These changes in gene expression will provide potential candidate genes that might be involved in scleral remodeling during axial elongation in myopia.
巩膜成纤维细胞参与近视轴向伸长过程中的巩膜重塑。机械负荷是基因表达的有力刺激因素。本研究旨在确定巩膜成纤维细胞基因表达对机械负荷的反应变化，并推测近视发展过程中巩膜重塑的可能机制。对人巩膜成纤维细胞（HSFs）进行30分钟和24小时的机械拉伸。采用基因微阵列分析来测量基因表达的变化。拉伸30分钟后，共有237个基因显示出表达上的差异和显著变化（P<0.01）。其中，28个未表达的基因开始表达（开启），而31个已表达的基因不再表达（关闭）。24小时后，308个基因显示出可重复的表达变化（P<0.01），同时29个基因开启，17个基因关闭。30分钟后，25个基因的表达至少有三倍的变化。这些基因包括细胞受体、蛋白激酶、细胞生长/分化因子、细胞外基质（ECM）蛋白、脂质代谢、蛋白质代谢、转录因子、结合蛋白和水通道的基因。24小时后，21个基因的表达至少有三倍的变化。这些基因包括细胞受体、蛋白激酶、细胞生长/分化因子、脂质代谢、ECM蛋白、转录因子和碳水化合物代谢的基因。RT-PCR和Southern印迹证实了所选基因表达的变化。在本研究中，我们在HSFs中鉴定出大量早期和晚期机械反应基因。这些基因表达的变化将提供可能参与近视轴向伸长过程中巩膜重塑的潜在候选基因。
