多西环素对体外培养的人视网膜色素上皮细胞移行的抑制作用  

Matrix metalloproteinases inhibitor doxycycline inhibits TGF-β1 induced RPE cell migration

作　　者：曾爱萍[1] 曾水清[1] 李鹏程[1] 程扬[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院眼科,武汉430022

出　　处：《中华眼科杂志》2007年第8期734-738,共5页Chinese Journal of Ophthalmology

基　　金：湖北省自然科学基金资助项目（2004ABA239）

摘　　要：目的探讨基质金属蛋白酶抑制剂多西环素对体外培养并经转化生长因子（TGF）-β1刺激的人视网膜色素上皮（RPE）细胞迁移的影响。方法对3～6代培养的人RPE细胞，用不同浓度（0．01、0，10、1．00、10．00μg／L）TGF-β1处理36h，采用明胶酶谱分析法检测人RPE细胞上清液中明胶酶的活性，采用Boyden室迁移测定法评估RPE细胞经TGF-β1刺激后的迁移情况。结果TGF-β1能促进人RPE细胞中基质金属蛋白酶一2的分泌，并具有浓度依赖性。在无多西环素的环境下，TGF-β1刺激RPE细胞后，能明显促进RPE细胞迁移，迁移的RPE细胞数约增加27％，与对照组相比差异有统计学意义（P〈0．01）。在多西环素的环境中，TGF-β1刺激人RPE细胞后迁移受到明显抑制，随着多西环素浓度的增加，RPE细胞穿透微小多孔膜发生迁移的细胞数相应减少，迁移的RPE细胞数约减少50％～70％，与对照组相比差异有统计学意义（P〈0．01）。结论多西环素能抑制TGF-β1作用所致的RPE细胞迁移，TGF-β1介导的基质金属蛋白酶-2活性的增加在RPE细胞的迁移中起重要作用。Objective To investigate the effects of matrix metalloproteinases （MMP） inhibitor （Doxyeyeline） on retinal pigment epithelial （RPE） cell migration induced by transforming growth factor beta 1 （TGF-β1 ）. Methods The third to sixth passage human RPE cells cultures were treated with TGF-β1 at different concentrations（0.01, 0.10, 1.00, 10.00 μg/L）,and the conditioned media were collected after 36 h of TGF-β1 exposure. Gelatinase activities in conditioned media were analyzed by zymography. Migration assay was performed in a Boyden chamber with addition of different concentrations of Doxycycline. The number of migrated cells was counted under light microscopy. Results RPE cells migration were stimulated by TGF-β1 significantly,the number of migrated RPE cells increased about 27% compared with the control （P 〈 O. 01 ）. In the presence of Doxycycline, the RPE cells migration induced by TGF-β1 was inhibited and the number of migrated cells was reduced to 50% - 70% compared with control （ P 〈 0.01 ）. TGF-β1 also stimulated the secretion of MMP-2 in dose-dependent manner. Conclusions This study indicates that Doxyeyeline can inhibit migration of RPE cells stimulated by TGF-β1. RPE migration induced by TGF-β1 may act partially through the stimulation of MMP production.

关 键 词：转化生长因子Β1 色素上皮 眼 明胶酶A 多西环素 细胞运动 

分 类 号：R686[医药卫生—骨科学]