转化生长因子β_1调节培养的人视网膜色素上皮细胞分泌基质金属蛋白酶-2

Regulation of Matrix Metalloproteinase-2 Production by TGF-β_1 in Human RPE Cells

目的研究转化生长因子β1(TGF-β1)对培养的人视网膜色素上皮(HRPE)细胞分泌基质金属蛋白酶-2(MMP-2)的调节作用.方法 3～4代培养的人视网膜色素上皮细胞,经不同浓度的TGF-β1(0.01、0.1、1、10 ng/ml)处理,36 h后收集条件培养基,或经TGF-β1作用不同时间(24、36、48 h)后收集条件培养基,采用明胶酶谱分析方法定量检测HRPE细胞上清液中MMP-2的活性水平.结果培养的HRPE细胞上清液中可检测到MMP-2,经TGF-β1处理后,能显著上调培养基中MMP-2的分泌.TGF-β1的浓度从0.01～10 ng/ml,MMP-2的分泌活性有明显的剂量依赖性,同时MMP-2的分泌活性随TGF-β1作用时间延长而增强. 结论培养的HRPE细胞能分泌MMP-2,TGF-β1能上调HRPE细胞中MMP-2的分泌.MMP-2分泌增加,可能在玻璃体视网膜的疾病中有利于HRPE细胞的迁移.

Objective To study the regulatory effects of transforming growth factor β1 （TGF-β1） on the secretion of matrix metalloproteinase-2 （MMP-2） by cultured human retinal pigment epithelial cells （RPE）. Methods The third-forth passage human RPE were treated with TGF β1 at different concentrations （0.01, 0. 1, 1, 10 ng/ml）, and the conditioned media were collected after 36 h, or for different time （24, 36, 48 h）, and the conditioned media were collected. The activities of MMP-2 in human RPE supernatant were quantitatively determined by gelatin zymography. Results MMP-2 in the conditioned medium was detected by zymography. The MMP-2 secretion was markedly up-regulated by TGF-β1. The MMP 2 activity was stimulated by TGF-β1 at 0.01, 0. 1, 1, 10 ng/ml in a dose-dependent manner, and the secretion of MMP-2 was time-dependently increased by TGF-β1. Conclusion Cultured human RPE can secret MMP-2. The production of MMP-2 is up-regulated by TGF-β1. Increased MMP-2 release may result in facilitated migration of RPE in the pathogenesis of vitreoretinopathy.