人眼巩膜成纤维细胞中视黄酸受体的分布及其对巩膜成纤维细胞的生长调控  

Expression of retinoid acid receptors in human scleral fibroblasts and regulation of growth of fibroblasts by retinoic acid

作　　者：闫东升[1] 周翔天[1] 陈晓燕[1] 吕帆[1] 王教[1] 胡诞宁[1,2] 瞿佳[1] 

机构地区：[1]温州医学院眼视光学院,325003 [2]温州医学院近视研究所纽约医学院纽约眼耳鼻喉科医院组织培养中心

出　　处：《中华眼科杂志》2007年第8期750-753,共4页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金资助项目（30371507,30500549）;浙江省自然科学基金资助项目（302095,11205739）;浙江省卫生厅重大课题资助项目（2002ZD009）;浙江省科技厅科技攻关计划一重点科研项目一社会发展项目（2003C23005）

摘　　要：目的研究正常人眼巩膜成纤维细胞视黄酸受体亚型的表达，进一步研究视黄酸对人巩膜成纤维细胞的生长调控作用，从而阐明其和近视发生发展的关系。方法应用定点解剖及游走促进法分离培养人巩膜成纤维细胞，取3～5代生长良好的细胞，运用逆转录聚合酶链式反应（RT—PCR）检测培养的巩膜成纤维细胞视黄酸受体亚型的表达；在培养液中分别加入0．01、0．10、1．00、10．00和100．00nmol／L的视黄酸，6d后进行细胞计数。每株细胞重复3次，共用3株细胞，观察视黄酸对巩膜成纤维细胞的生长调控作用。结果培养的巩膜成纤维细胞呈双极型或纺锤型，平均分裂时间为2～3d左右，细胞生长旺盛，近融合时细胞成规则排列的纺锤型。RT—PCR提示体外培养的人巩膜成纤维细胞表达所有的视黄酸受体亚型。视黄酸能明显抑制人巩膜成纤维细胞的生长，对细胞生长的抑制作用呈明显的剂量效应关系，RA浓度≥0．10nmol／L时，与无RA的对照组相比较，差异有统计学意义（P〈0．05）。结论所有的视黄酸受体亚型在人巩膜成纤维细胞上均有分布，视黄酸能明显抑制人巩膜成纤维细胞的生长。Objective The aim of this study was to identify the presence of retinoid acid receptors in cultured human scleral fibroblasts and the effect of retinoic acid （RA） on the regulation of cell growth. Methods Human scleral fibroblasts from three donors were isolated and cultured in Dulbecco Modified Ealge＇s Medium. Cells from the third or fifth generation were used in the present study. Reverse Transcription-Polymerase Chain Reaction （RT-PCR） was used to detect the expression of retinoid acid receptor subtypes in the fibroblasts. RA was added to the culture medium at concentrations of 0. 01,0. 10, 1.00,10. 00 and 100. 00 nmol/L, separately. Cells were counted and statistically analyzed six days after being treated with RA. Each strain of the cells was examined three times. Results Cultured fibroblasts mainly showed a bipolar or spindle-shaped morphology with an average time of 2 to 3 days for each division. RT-PCR showed mRNA expression of six subtypes of RA receptors （RARα, RARβ, RARγ, RXRα, RXRβ, and RXRγ）. The proliferation of scleral fibroblasts was inhibited by 2. 2%, 17. 6%, 37. 6%, 43. 8% and 63. 2% following the treatment with 0. 01, 0. 10, 1.00,10. 00 and 100. 00 nmol/L of RA, respectively. Cell proliferation in RA-treated samples reduced significantly as compared to the control samples （P 〈 0. 05 ） when the RA concentration exceeded 0. 10 nmol/L. Conclusions Six retinoid acid receptor subtypes are present in cultured human scleral fibroblasts and the growth of scleral fibroblasts was inhibited significantly by retinoic acid. These findings indicate that the sclera is a potential site of action for the retinoid acid during progression of myopia.

关 键 词：巩膜 成纤维细胞 受体 维甲酸 近视 

分 类 号：R686[医药卫生—骨科学]