摘要
目的研究糖基化终末产物(AGEs)对人视网膜色素上皮(RPE)细胞表达基质金属蛋白酶-9(MMP-9)的影响。方法体外培养人RPE细胞,加入质量浓度为100mg/L的AGEs分别作用不同时间(6、12、24、36h),采用免疫细胞化学法检测MMP-9蛋白的表达,应用逆转录聚合酶链式反应(RT-PCR)法检测MMP-9 mRNA水平。结果免疫细胞化学染色结果表明,对照组仅少量细胞有MMP-9弱阳性表达,而实验组各时间段均可见细胞浆黄色或棕黄色着染,随作用时间的延长其表达逐渐增强。各时间段两两结果比较,差异均有统计学意义(P<0.05)。RT-PCR检测显示MMP-9 mRNA量与MMP-9蛋白表达变化一致。结论AGEs能促使人RPE细胞MMP-9表达上调,在一定范围内与作用时间有关。
Objective Diabetic retinopathy is a common and serious microvascular complication. Recent studies have found that advanced glycation end products （AGEs） in the retinal blood vessel is an important cause in the development of diabetic retinopathy. The aim of this study was to investigate the effects of AGEs on the expression of matrix metalloproteinase 9 in human retinal pigment epithelial （RPE） cells. Methods Human RPE cells lines was maintained in DMEM/F12 medium containing 10% fetal bovine serum to prepare the cell suspension with the RPE cells 5 × 10^4/mL. Passaged human RPE cells were exposed with or without 100 mg/L AGEs for different time of period（6,12,24,36 hours）. The expression of MMP-9 protein was assessed by immunocytochemistry assay,and the MMP-9 mRNA was studied by reverse transcriptase PER. Results MMP-9 positive cells showed a brown-yellow staining in cytoplasm. Only small numbers of weaker positive cells for MMP-9 were seen in the control group in different cultured time,but in experimental groups,the intensity of positive staining for MMP-9 was obviously increased. MMP-9 expression peaked in 6 hours and gradually decreased at 12,24 and 36 hours after addition of AGE and showed significant differences between control group and AGEs groups in various time points （P 〈0. 05）. The changes in MMP-9 mRNA paralleled to the changes of MMP-9 protein. Conclusion AGEs could up-regulate the expression of MMP-9 in human RPE cells at a time-dependent manner.
出处
《眼科研究》
CSCD
北大核心
2008年第4期253-256,共4页
Chinese Ophthalmic Research
