Li Chuanxu, McFadden Sally A, Morgan Ian, Cui Dongmei, Hu Jianmin, Wan Wenjuan, Zeng Junwen
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Mol Vis. 2010 Apr 15;16:689-97.
Fibulin-1 (FBLN1) mRNA is expressed in human sclera and is an important adhesion modulatory protein that can affect cell-matrix interactions and tissue remodeling. Scleral remodeling is influenced by all-trans retinoic acid (RA). Our purpose was to confirm the presence of fibulin-1 protein in guinea pig sclera and investigate the effect of RA on the expression of fibulin-1 in guinea pig sclera in vivo and in cultured human scleral fibroblasts (HSFs).
Confocal fluorescence microscopy was used to study fibulin-1 and aggrecan expression and localization in sclera from control guinea pigs and in animals given RA by daily gavage from 4 to 8 days of age. The effects of RA (from 10(-9) to 10(-5) M) on fibulin-1 expression in HSFs were observed by immunohistochemistry and assayed by real-time PCR and western blot analysis.
Fibulin-1 protein expression was detected by confocal fluorescence microscopy in guinea pig sclera and in cultured HSFs. Upregulation of fibulin-1 in scleral tissue was observed after feeding with RA. In vitro, the level of Fbln1 mRNA was increased after treatment of HSFs with RA (at concentrations of 10(-8) to 10(-6) M; p<0.001), with a maximum effect at 10(-7) M. Fibulin-1 protein levels were significantly increased after treatment of HSFs with 10(-7) M of RA for 24 or 48 h (p<0.05).
Fibulin-1 protein was expressed in guinea pig sclera and cultured HSFs. Expression was regulated by RA, a molecule known to be involved in the regulation of eye growth. Further studies on the role of fibulin-1 in the regulation of eye growth, including during the development of myopia, are therefore warranted.
纤维连接蛋白-1（FBLN1）mRNA在人巩膜中表达，是一种重要的黏附调节蛋白，可影响细胞与基质的相互作用以及组织重塑。巩膜重塑受全反式维甲酸（RA）影响。我们的目的是证实豚鼠巩膜中纤维连接蛋白-1蛋白的存在，并研究RA对豚鼠巩膜及培养的人巩膜成纤维细胞（HSFs）中纤维连接蛋白-1表达的影响。
采用共聚焦荧光显微镜研究对照组豚鼠巩膜以及4至8日龄每日经口灌胃给予RA的动物巩膜中纤维连接蛋白-1和聚集蛋白聚糖的表达及定位。通过免疫组织化学观察RA（浓度为10⁻⁹至10⁻⁵M）对HSFs中纤维连接蛋白-1表达的影响，并通过实时PCR和蛋白质印迹分析进行检测。
通过共聚焦荧光显微镜在豚鼠巩膜和培养的HSFs中检测到纤维连接蛋白-1蛋白表达。给予RA后，观察到巩膜组织中纤维连接蛋白-1上调。在体外，用RA（浓度为10⁻⁸至10⁻⁶M）处理HSFs后，Fbln1 mRNA水平升高（p<0.001），在10⁻⁷M时作用最大。用10⁻⁷M的RA处理HSFs 24或48小时后，纤维连接蛋白-1蛋白水平显著升高（p<0.05）。
纤维连接蛋白-1蛋白在豚鼠巩膜和培养的HSFs中表达。其表达受RA调节，RA是一种已知参与眼睛生长调节的分子。因此，有必要进一步研究纤维连接蛋白-1在眼睛生长调节中的作用，包括在近视发展过程中的作用。
